LOCUS       sequence-254033-        2710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Module cloning vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2710)
  AUTHORS   Goulev Y, Matifas A, Heyer V, Reina-San-Martin B, Charvin G
  TITLE     COSPLAY: An expandable toolbox for combinatorial and swift 
            generation of expression plasmids in yeast.
  JOURNAL   PLoS One. 2019 Aug 28;14(8):e0220694. doi: 
            10.1371/journal.pone.0220694. eCollection 2019.
  PUBMED    31461461
REFERENCE   2  (bases 1 to 2710)
  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: "PLoS One.";
            date: "2019-08-28"; volume: "14(8):e0220694. doi"; pages: " 
            10.1371/journal.pone.0220694. eCollection 2019"
COMMENT     Sequence Label: pUC57BsaI
FEATURES             Location/Qualifiers
     source          1..2710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     117..134
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    251..272
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(287..292,293..310,311..317)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    325..341
                     /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)."
     primer_bind     330..352
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     349..365
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     349..365
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             361..708
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHAGLCSRRARDPISRCIREVPSSNSLAVVLQRRDWEN
                     PGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIW
                     CTLSTICSDAA"
     primer_bind     complement(459..476)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(459..475)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(468..490)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(684..703)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     803..825
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(863..881)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        949..1053
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1054..1122,1123..1914)
                     /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     complement(1272..1291)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2085..2673
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2574..2593
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
       61 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
      121 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
      181 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
      241 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
      301 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
      361 atgaccatga ttacgccaag cttgcatgca ggcctctgca gtcgacgggc ccgggatccg
      421 atatctagat gcattcgcga ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt
      481 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
      541 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
      601 ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
      661 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
      721 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
      781 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
      841 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
      901 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
      961 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     1021 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     1081 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     1141 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     1201 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     1261 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     1321 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     1381 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     1441 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     1501 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     1561 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     1621 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     1681 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     1741 gctgataaat ctggagccgg tgagcgtgga tctcgcggta tcattgcagc actggggcca
     1801 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     1861 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     1921 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     1981 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     2041 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2101 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2161 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     2221 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     2281 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     2341 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     2401 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     2461 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     2521 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     2581 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     2641 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     2701 ttcctggcct
//
