LOCUS       sequence-200872-        3853 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SapTrap destination vector for building combined sgRNA expression + 
            repair template vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3853)
  AUTHORS   Schwartz ML, Jorgensen EM
  TITLE     SapTrap, a Toolkit for High-Throughput CRISPR/Cas9 Gene Modification
            in Caenorhabditis elegans.
  JOURNAL   Genetics. 2016 Feb 2. pii: genetics.115.184275.
  PUBMED    26837755
REFERENCE   2  (bases 1 to 3853)
  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: "Genetics. 
            2016 Feb 2. pii: genetics.115.184275."
COMMENT     Sequence Label: pMLS256
FEATURES             Location/Qualifiers
     source          1..3853
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        125..624
                     /label=CeU6 promoter
                     /note="promoter for the C. elegans K09B11.12 U6 snRNA gene 
                     (Friedland et al., 2013)"
     misc_RNA        679..754
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    762..998
                     /label=3' CeU6
                     /note="region downstream of the C. elegans K09B11.12 U6 
                     snRNA gene"
     primer_bind     complement(1005..1022)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1176..1764)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1256..1275)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1935..2726,2727..2795))
                     /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     2558..2577
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2796..2900)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2926..3381)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3063..3084)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3275..3294
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(3394..3765)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSTAVAANSIGEEPMDPLVIWNSAGL
                     LFGNQAYRYRRPRGGARYPIRPIVSRITRAHWPSFYNVVTGKTLALPNLIALQHIPLSP
                     AGVIAKRPAPIALPNSCAA"
     primer_bind     3508..3530
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3522..3539
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3523..3539
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3549..3568
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3549..3567
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3592..3608
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3593..3609
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3722..3742)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3722..3740)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3761..3777)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3761..3777)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3774..3796)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3785..3801
                     /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(3809..3815,3816..3833,3834..3839))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
       61 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
      121 tgggctccaa gaactcgtac aaaaatgctc tgaagtaggt ctcgagatca atactgggaa
      181 gacgaaagtc ttgcgaaacc gattcgctga ccccagtaaa gtctacttcg gtagcccttc
      241 ccccaccacc cagctcgacg acgtcgacga gtacatctac ctcggtcgtc aaatcaacgc
      301 ccaaaacaac ttgatgccgg aaatccaccg aagacgtcga gcagcctggg ctgcattcaa
      361 tggaatcaag aatgccaccg actccatcac cgacaagaag attcgtgcga atctgttcga
      421 ctcaattgtc cttccagcgc tcacctacgt ttcagaagcc tggacattca ccaaagctct
      481 atccgaacga gtacgaatca cacatgcctc ccccacaaat tattgatcta catccccatt
      541 tggtatagtg tcttacagga ctctcaatat gtcagctgca tgataaatgg tccctatata
      601 attgaattgc aaatctaaat gtttgcgaag agcaggcctg cagaattcca tatgactagt
      661 ggatccatgg gctcttcggt tttagagcta gaaatagcaa gttaaaataa ggctagtccg
      721 ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt tcaataatat tctagatatc
      781 cctttttgaa actaaaatga aatactctaa atttttgagt tcaatgtttc aactaatctt
      841 ggttaaaggt aaattcccgg agggacgatt ttaaagttgc cagagttgat taaattggtc
      901 caaaaaattt tattaggaaa ctcgaaaaaa aattgatttt ttttgacaat attccaaaga
      961 aaattacaaa atctcgacgc caaacatagt cggctgtgcg cttcctcgct cactgactcg
     1021 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     1081 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     1141 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     1201 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     1261 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     1321 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     1381 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     1441 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     1501 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     1561 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     1621 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     1681 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     1741 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     1801 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     1861 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     1921 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     1981 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     2041 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     2101 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     2161 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     2221 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     2281 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     2341 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     2401 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     2461 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     2521 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     2581 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     2641 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     2701 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     2761 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     2821 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     2881 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     2941 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     3001 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     3061 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     3121 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3181 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3241 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3301 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3361 atgcgccgct acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc
     3421 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     3481 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     3541 agcgcgcgta atacgactca ctatagggcg aattgggtac cgggcccccc ctcgaggtcg
     3601 acggtatcga taagcttgat taccgaagag caggcctgca gaattccata tgactagtgg
     3661 atccatgggc tcttcgccaa tcgaattggc cgccaccgcg gtggagctcc agcttttgtt
     3721 ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt
     3781 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3841 cctggggtgc cta
//
