LOCUS       sequence-497165-        5388 bp    DNA     circular SYN 13-JAN-2026
DEFINITION  P7SulA.1 tagging casette for WTC847 with Hygromycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5388)
  TITLE     Stelling Lab Plasmids 2025
REFERENCE   2  (bases 1 to 5388)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 13, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: P2755
FEATURES             Location/Qualifiers
     source          1..5388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     167..186
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     400..422
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     414..431
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     415..431
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     438..457
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        438..456
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     gene            516..2091
                     /label=hphMX6
                     /note="yeast selectable marker conferring hygromycin 
                     resistance"
     promoter        516..859
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             860..1888
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      1894..2091
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             2881..2952
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     promoter        complement(3015..3033)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3054..3070)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3054..3070)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3067..3089)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3078..3094
                     /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(3102..3108,3109..3126,3127..3132))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3147..3168
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3285..3302)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3456..4044)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3536..3555)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4215..5006,5007..5075))
                     /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     4838..4857
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5076..5180)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(5206..5388,1..273))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5343..5364)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
       61 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
      121 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
      181 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
      241 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
      301 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
      361 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
      421 cgacggccag tgaattgtaa tacgactcac tatagggcga attggagctc gatcctcgag
      481 agcttgcctt gtccccgccg ggtcacccgg ccagcgacat ggaggcccag aataccctcc
      541 ttgacagtct tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg tacatttagc
      601 ccatacatcc ccatgtataa tcatttgcat ccatacattt tgatggccgc acggcgcgaa
      661 gcaaaaatta cggctcctcg ctgcggacct gcgagcaggg aaacgctccc ctcacagacg
      721 cgttgaattg tccccacgcc gcgcccctgt agagaaatat aaaaggttag gatttgccac
      781 tgaggttctt ctttcatata cttcctttta aaatcttgct aggatacagt tctcacatca
      841 catccgaaca taaacaacca tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa
      901 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga
      961 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
     1021 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatcgg ccgcgctccc
     1081 gattccggaa gtgcttgaca ttggggagtt cagcgagagc ctgacctatt gcatctcccg
     1141 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
     1201 gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat cttagccaga cgagcgggtt
     1261 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
     1321 gattgctgat ccccacgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc
     1381 cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca
     1441 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
     1501 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
     1561 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
     1621 tccggagctt gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg gtcttgacca
     1681 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
     1741 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
     1801 cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc
     1861 cagcactcgt ccgagggcaa aggaataatc agtactgaca ataaaaagat tcttgttttc
     1921 aagaacttgt catttgtata gtttttttat attgtagttg ttctatttta atcaaatgtt
     1981 agcgtgattt atattttttt tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg
     2041 cagaaagtaa tatcatgcgt caatcgtatg tgaatgctgg tcgctatact gctgtcgatt
     2101 cgatactaac gccgccatcc agtgtttaaa cgaactagtg cggccgccag ttcgagttta
     2161 tcattatcaa tactgccatt tcaaagaata cgtaaataat taatagtagt gattttccta
     2221 actttattta gtcaaaaaat tagccctgta catccataca gtgtctgggt gactgtacat
     2281 ccatacagca tccactaatt ttaattctgc tgtaacccgt acatgcccaa aatagggggc
     2341 gggttacaca gaactgtaca tccatacagt gtctgggtga ctgtacatcc atacagcatc
     2401 cactaaatat aatggagcct gtacatccat acagcatcca ctaaaaaaaa gaatcccagc
     2461 accaaaatat tgttttcttc accaaccatc agttcatagg tccattctct tagcgcaact
     2521 acagagaaca ggggcacaaa caggcaaaaa acgggcacaa cctcaatgga gtgatgcaac
     2581 ctgcctggag taaatgatga cacaaggcaa ttgacccacg catgtatcta tctcattttc
     2641 ttacaccttc tattaccttc tgctctctct gatttggaaa aagctgaaaa aaaaggttga
     2701 aaccagttcc ctgaaattat tcccctctgt acatccatac agtataaata ctgtacatcc
     2761 atacagtaat tctgtaaatc tatttcttaa acttcttaaa ttctactttt atagttagtc
     2821 ttttttttag ttttaaaaca ccaagaactt agtttcgaat aaacacacat aaacaaaatg
     2881 gattataaag atgacgatga caaggattat aaagatgacg atgacaagga ttataaagat
     2941 gacgatgaca agggaggtgg tggtggtgga ggcggtccga cagagaatca tcgatgggta
     3001 cccagctttt gttcccttta gtgagggtta atttcgagct tggcgtaatc atggtcatag
     3061 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3121 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     3181 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3241 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3301 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3361 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3421 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3481 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3541 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3601 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3661 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3721 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3781 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3841 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3901 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3961 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4021 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4081 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4141 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4201 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4261 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4321 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4381 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4441 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     4501 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4561 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4621 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4681 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4741 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4801 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4861 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4921 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4981 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5041 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5101 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5161 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     5221 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     5281 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     5341 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacg
//
