LOCUS       sequence-345248-       10493 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targeting vector for generating Wnk1 STREAMING-tag KI cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10493)
  AUTHORS   Ohishi H, Shimada S, Uchino S, Li J, Sato Y, Shintani M, Owada H, 
            Ohkawa Y, Pertsinidis A, Yamamoto T, Kimura H, Ochiai H
  TITLE     STREAMING-tag system reveals spatiotemporal relationships between 
            transcriptional regulatory factors and transcriptional activity.
  JOURNAL   Nat Commun. 2022 Dec 20;13(1):7672. doi: 10.1038/s41467-022-35286-2.
  PUBMED    36539402
REFERENCE   2  (bases 1 to 10493)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2022-12-20"; volume: "13(1):7672. doi"; pages: " 
            10.1038/s41467-022-35286-2"
COMMENT     Sequence Label: pTV-Wnk1-5prime-1000-24MS96T-3F_NeoR
FEATURES             Location/Qualifiers
     source          1..10493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2157..2166
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2469..2487
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2498..2516
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2527..2545
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2572..2590
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2601..2619
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2630..2648
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2675..2693
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2704..2722
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2733..2751
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2778..2796
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2807..2825
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2836..2854
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2881..2899
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2910..2928
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2939..2957
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2984..3002
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3013..3031
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3042..3060
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3087..3105
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3116..3134
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3145..3163
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3190..3208
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3219..3237
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3248..3266
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3293..3311
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3322..3340
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3351..3369
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3396..3414
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3425..3443
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3454..3472
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3499..3517
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3528..3546
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3557..3575
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3602..3620
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3631..3649
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3660..3678
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3705..3723
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3734..3752
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3763..3781
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3808..3826
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3837..3855
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3866..3884
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3911..3929
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3940..3958
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3969..3987
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4014..4032
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4043..4061
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4072..4090
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4117..4135
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4146..4164
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4175..4193
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4220..4238
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4249..4267
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4278..4296
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4323..4341
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4352..4370
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4381..4399
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4426..4444
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4455..4473
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4484..4502
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4529..4547
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4558..4576
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4587..4605
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4632..4650
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4661..4679
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4690..4708
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4735..4753
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4764..4782
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4793..4811
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4838..4856
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4867..4885
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4896..4914
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4941..4959
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4970..4988
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4999..5017
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5044..5062
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5073..5091
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5102..5120
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5147..5165
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5176..5194
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5205..5223
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5250..5268
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5279..5297
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5308..5326
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5353..5371
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5382..5400
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5411..5429
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5456..5474
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5485..5503
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5514..5532
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5559..5577
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5588..5606
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5617..5635
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5662..5680
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5691..5709
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5720..5738
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    5805..5830
                     /label=SA
                     /note="splice acceptor site"
     CDS             6196..6219
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6235..6258
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6274..6297
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(8313..8333)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8313..8331)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8352..8368)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8352..8368)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8365..8387)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8376..8392
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(8400..8406,8407..8424,8425..8430))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8445..8466
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8576..8593)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8747..9335)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8827..8846)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9506..10297,10298..10366))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     10129..10148
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10367..10457)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggtttgaca
      661 tgcctgcaag tgcaggtact cccttcttcg ctgacagaag gcgcctagtg gggtgactgc
      721 tctttttctc tccctctccc cctcctccca gttctttcgg tctcctctcc tcctggccag
      781 agaagtagcg gagcccgggc cccgcggtga gcggccctcc cctccccaca gttccctccc
      841 tcctctgccc ccggcgccgg ccagagaggg gaagaggttc gccaggcccg gggcgggcgg
      901 ggaggcctcg gggacggggg gggccgctcc tcaggcttcg aggcttcgag gcttcggccc
      961 tcagcatcta gtgatgggcg ccctctgaga ctggtcccct ctccggggcg cgtgctgcct
     1021 gaggcggcgg ccgagcgagc gagcgagcga gcgagcgacc gggaacccgg gccgcggccc
     1081 tccttcccct gcgcggcccc tcccactcct gtgccccggg ctgccaccgc ccggcgtcgg
     1141 ggctcgtccc tcgcgcgctg tgcctccgcc ctcagggcct atcccgccgt gctcggcggg
     1201 cgggcggcga cggcggcagc ggcagcggag ggagcttagc cgtcttcact tgggtcgcat
     1261 cctctcgccc aggagccaat tccgcgccct gctcctcatg actgcggcgc ttctgctgcc
     1321 ccagccctac cggcctccgc tcgctgcagg atggatgcgg actgtgcggc gctaaccccc
     1381 gtggctccgc tccctcctag cccgccgacg agcccgcccc gtgagccgca gcagcttcgg
     1441 tgccagcagc agcagccgcc gcagcagccg gactcagtgt cccgcgtcgc cctcggggcg
     1501 ccttgtcagt gctgagtggg agtcgtccgg tggttcggag cccgccgtag gagcgctgcc
     1561 cagcacttct tggtggcttc tccgggtcac ccttttccct tttcattcat aaatccgagc
     1621 ccgcattctt cactccatcg gaccgaccat gtctgacggt gccgccgaga agcaaagcca
     1681 ggtaagcgat aagtattcga tcatgcattg gttagcggcc gcgaattcgc ccttggatcc
     1741 ggaatgacca ccaggcattc cgatgccacg atggaccatc aggccatcga gctgcagaag
     1801 tgacgaccac gcacttcgga gtgacagagg aggatcaccc ctctggccac cagagtagag
     1861 catcagccta ctcacagagc ctacggagga ccaccccgta ggagtagagc gaggagcacc
     1921 agccctcgcg tgtgacgcac gagcatcagc cgtgcgatgg taagatccgg aatgaccacc
     1981 aggcattcct ccaaggcgat ggaccatcag gccatcgagg actcgaagtg acgaccacgc
     2041 acttcgtgaa ggcagaggag gatcacccct ctgcccacag gagtagagca tcagcctact
     2101 ccaagcagct acggaggacc accccgtaga agtagggcga ggagcaccag ccctcgccat
     2161 ggtggcacga gcatcagccg tgcagggtat agatccggaa tgaccaccag gcattcctgg
     2221 ccaacgatgg accatcaggc catcggcata gcgaagtgac gaccacgcac ttcagggtag
     2281 cagaggagga tcacccctct ggccacacga gtagagcatc agcctactcc acaggactac
     2341 ggaggaccac cccgtagaaa gtgggcgagg agcaccagcc ctcgccgtga tggcacgagc
     2401 atcagccgtg cgttaggaag atctcgattc gatgcagcct agttaggggt cgactagtct
     2461 actcactatc tctatcactg atagggaaac cttcctctct ctatcactga tagggacctt
     2521 ttcccttctc tatcactgat agggacccat attctcctag tttaacttct atctctatca
     2581 ctgataggga aactcacttc tctctatcac tgatagggac cttatcccct ctctatcact
     2641 gatagggata ttatatttcc tagtattcac ctcctctcta tcactgatag ggattcacca
     2701 ttttctctat cactgatagg gatatttcct tctctctatc actgataggg acatattttt
     2761 ccctagtcaa tcctcactct ctatcactga tagggactaa caaaattctc tatcactgat
     2821 agggaaaacc aaacatctct atcactgata gggatcatca ttttcctagt ataaccatac
     2881 tctctatcac tgatagggat cacccccact ctctatcact gatagggatt acacaacttc
     2941 tctatcactg atagggactc tctttatcct agtcacatta acatctctat cactgatagg
     3001 gacacctttt catctctatc actgataggg acatccatta ttctctatca ctgataggga
     3061 cattaactct cctagtaaaa tccccatctc tatcactgat agggaaacct catcatctct
     3121 atcactgata gggaatctat tccatctcta tcactgatag ggatatcttt tatcctagtc
     3181 ctaacatcat ctctatcact gatagggaac ccaaactttc tctatcactg atagggacca
     3241 atttacctct ctatcactga tagggaatct tcttatccta gtacaaaact aatctctatc
     3301 actgataggg aatccctccc ttctctatca ctgataggga tacctttcct tctctatcac
     3361 tgatagggaa ttacttttac ctagtaatac ctcaatctct atcactgata gggaaaatca
     3421 ctaatctcta tcactgatag ggatttttat tattctctat cactgatagg gaactccttt
     3481 aacctagtct attccatttc tctatcactg atagggattc actaccctct ctatcactga
     3541 tagggattat ttcttctctc tatcactgat agggaatttt tccttcctag taccataacc
     3601 ctctctatca ctgataggga acaccaatat tctctatcac tgatagggac attatttatt
     3661 ctctatcact gatagggaat tattactccc tagttcttac attatctcta tcactgatag
     3721 ggaacccctc tcctctctat cactgatagg gatatcaatc actctctatc actgataggg
     3781 acctattata ccctagtcca atatatatct ctatcactga tagggaccac acttcttctc
     3841 tatcactgat agggatcacc ttatatctct atcactgata gggacactct ctatcctagt
     3901 ccaatccttt tctctatcac tgatagggat aaacacatct ctctatcact gatagggata
     3961 cattttactc tctatcactg atagggattc tccctttcct agtcttaact tcttctctat
     4021 cactgatagg gatttttcca attctctatc actgataggg aatcacttca ttctctatca
     4081 ctgataggga ttatttctcc cctagttcac tcctactctc tatcactgat agggactccc
     4141 aatattctct atcactgata gggatatttt tctctctcta tcactgatag ggacacctcc
     4201 tctcctagtt accacaactt ctctatcact gatagggaat actctcaatc tctatcactg
     4261 atagggattt ctatttctct ctatcactga tagggaatac cctctaccta gtaacaaatc
     4321 tttctctatc actgataggg aataatatat ctctctatca ctgataggga tcttataccc
     4381 tctctatcac tgatagggac cttccacctc ctagttaatc aaatatctct atcactgata
     4441 gggataatct actttctcta tcactgatag ggaaactcat ccatctctat cactgatagg
     4501 gatttttaac ttcctagtat ccattctatc tctatcactg atagggataa tacaaactct
     4561 ctatcactga tagggatact ctcctttctc tatcactgat agggacctta ttttccctag
     4621 tataatccct atctctatca ctgataggga acattaacta tctctatcac tgatagggat
     4681 tttattcctt ctctatcact gatagggaac ttacctatcc tagtataccc ataatctcta
     4741 tcactgatag ggaaaccaaa atttctctat cactgatagg gaatccaatt actctctatc
     4801 actgataggg atttctatct ccctagttta aaaccaatct ctatcactga tagggaaccc
     4861 ctaaactctc tatcactgat agggatttct cttattctct atcactgata gggaccttct
     4921 ctcacctagt ctactcacta tctctatcac tgatagggaa accttcctct ctctatcact
     4981 gatagggacc ttttcccttc tctatcactg atagggaccc atattctcct agtacactat
     5041 aaatctctat cactgatagg gaaaacatac cttctctatc actgataggg actatatttt
     5101 ttctctatca ctgataggga tctaatactt cctagtactt taaatatctc tatcactgat
     5161 agggaatcta ctaaatctct atcactgata gggacccata aacttctcta tcactgatag
     5221 ggatattctt taacctagta caataccatt ctctatcact gatagggacc cccctaattc
     5281 tctatcactg atagggaatt tttttcctct ctatcactga tagggattat ctcttcccta
     5341 gttcctctta aatctctatc actgataggg atctcactac ttctctatca ctgataggga
     5401 aactcataac tctctatcac tgatagggaa aattatcttc ctagtaaaac acctatctct
     5461 atcactgata gggatattat accatctcta tcactgatag ggatttatca ttttctctat
     5521 cactgatagg gatttttacc atcctagtta aaaatatctc tctatcactg atagggatca
     5581 ctaactctct ctatcactga tagggaacta ttctattctc tatcactgat agggacactt
     5641 tttaacctag tacattaccc atctctatca ctgataggga acctatctct tctctatcac
     5701 tgatagggat attttttcat ctctatcact gatagggaaa actttacacc taggattacc
     5761 ctgttatccc tactcgacat cgattagctg atacgataaa gcttctgacc tcttctcttc
     5821 ctcccacagg gatgtctgag ctacgataca gagatcctca cagttgagta cggatttctg
     5881 cccattggca aaatagtcga ggaaaggatt gaatgcacag tttacacagt ggacaagaat
     5941 ggctttgtct atacacagcc aatagcccag tggcataacc gaggagagca agaggtattc
     6001 gagtattgtc tcgaagatgg gagtatcatc cgagccacca aagaccacaa attcatgaca
     6061 acagacggac aaatgctgcc tattgacgaa atatttgaga gaggtttgga tctgaagcag
     6121 gttgacgggc tgcccgccgg agccggtgcc ggagctgcaa gcgggaaaga gaaaacacat
     6181 gtttcccggc ctagggatta caaggatgac gacgataagg gcactgcaag cagtgactat
     6241 aaggatgatg atgacaaagg gtctggtggg ggcgactata aggatgacga tgacaagcgg
     6301 cctaggctta atagcaacat ggacgcagac ctttatggat ataagtgggc tcgcgacaat
     6361 gtaggacaga gcggcgctac tatttatcgg ctctatggaa aacccgatgc tcccgagctg
     6421 ttcctgaagc acggcaaggg atccgtagca aacgacgtta cagacgaaat ggtacgactc
     6481 aactggttga cagaatttat gcccctgccc actatcaagc acttcataag aactcctgat
     6541 gacgcttggc tgctgacaac cgcaatcccc ggtaaaacag cattccaggt ccttgaagag
     6601 tatccagact ctggtgagaa tattgtggac gcactggcag tattcctcag gcggctccac
     6661 agtattcctg tttgcaattg tccctttaac tctgacaggg tattcaggct cgcccaggcc
     6721 caatcacgaa tgaataatgg cttggttgat gcaagcgatt tcgatgatga acgcaatggc
     6781 tggcctgttg aacaagtttg gaaagaaatg cacaagctcc tgcccttttc acccgattct
     6841 gtagtgactc acggggactt cagcctcgac aatttgatct tcgatgaagg aaagcttatc
     6901 ggctgcatag atgttgggcg cgttgggata gccgacagat accaggacct ggccatattg
     6961 tggaattgtc tcggcgagtt ttcccctagc ttgcaaaaac ggctctttca gaagtatggc
     7021 atagataacc ccgacatgaa taaacttcaa ttccatctga tgcttgacga gttcttcgga
     7081 actgctggag ctggagctgg ggctgtaaag ggaggaggtg gatctggtgg aggaggtagt
     7141 ggaggaggtg gaagtggtgg gggagtaaag atcatcagtc gaaaatccct gggcacccaa
     7201 aatgtatacg atattggggt cgagaaagat cacaattttc tcctgaagaa tggtctggtt
     7261 gcaagcaatt gcttcggcac tccgggcttc cttacgcctc cggcgcccgt ccccaagaat
     7321 ggctcaagct ccgattcttc cgtgggcgag aaactggggg ctacggtcgc cgactcggga
     7381 gttggcagga cggaggaata tcggcgccgc cggcacacta tggacaagga cagtcgtggg
     7441 gcggccgcca ccaccacccc cacggagcac cgattcttcc gccgaagcgt catctgcgac
     7501 tccaatgcca ccgcgttgga gctgccgggc ctccctctct ctatcccgca acctagtgta
     7561 cctgcagtgg tgccccagag tgctccaccc gaacctcaca gagaggaaac tttgaccgcc
     7621 accgttgctt cccaggtgtc tcagcaaccc tcggctgctg cctcccctgg ggaacaagcc
     7681 gttgtaggct cggccaccac gaccgtcccc agcagtacca gcaaagaccg cccggtgtcc
     7741 cagcccagcc tcgtggggag caaagaggag ccaccgccgt ccagaagtgg aagtggcagt
     7801 ggtggcgcca gcgccaaaga ggcgcaggag gataggagcc agcaacagga tgatatcgaa
     7861 gagctggaga ccaaggcagt gggaatgtcc aatgacggtc gctttctcaa atttgacatc
     7921 gaaatcggca gaggctcctt taagacagtc tacaaaggtc tggacaccga aaccactgtg
     7981 gaagtcgcct ggtgtgaatt gcaggtaatg ccccaccccc ttttttctta ttcggtcgtt
     8041 tggatcccaa atttggctct gttaagccag tttgtcgagt tcactcttta cttgtaattc
     8101 tctgactctg agagggcggg aagactagtg tccaaagagg ttgaaattgg gagattgtgg
     8161 agcttgtagt tagttaggtg tttggaaaag ataggaacta ttggtgtcaa gaaggattgt
     8221 tatcagccag gggtaaaaag agtcatttaa gtacaaactt gtgtgtttaa tgtcagcctg
     8281 cacttgcagg catgtcaaac cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     8341 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     8401 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     8461 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     8521 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcc gcttcctcgc
     8581 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     8641 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     8701 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     8761 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     8821 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     8881 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     8941 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     9001 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     9061 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     9121 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     9181 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     9241 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     9301 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     9361 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     9421 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     9481 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     9541 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     9601 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcggctg ccacgctcac
     9661 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     9721 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     9781 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     9841 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     9901 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     9961 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
    10021 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
    10081 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
    10141 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
    10201 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
    10261 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
    10321 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttccttttca
    10381 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
    10441 ttagaaaaat aaacaaatag ggattccgcg cacatttccc cgaaaagtgc cac
//
