LOCUS       sequence-252809-        2770 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Position 1 module- Linker .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2770)
  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 2770)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pMV8
FEATURES             Location/Qualifiers
     source          1..2770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    102..123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(138..143,144..161,162..168)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    176..192
                     /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     181..203
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     200..216
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     200..216
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(280..303)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      303..312
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(370..387)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(370..386)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(379..401)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(595..614)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     714..736
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(774..792)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        860..964
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(965..1033,1034..1825)
                     /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(1183..1202)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1996..2584
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2485..2504
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2738..2755
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
       61 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
      121 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
      181 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcttgcatgc
      241 aggcctctgc agtcgacggg cccggtctct catacctccc ttgtcatcgt cgtccttgta
      301 gtccatggtg gcggatcgag accgggatcc gatatctaga tgcattcgcg aggtaccgag
      361 ctcgaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
      421 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
      481 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta
      541 ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat
      601 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
      661 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
      721 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
      781 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
      841 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
      901 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
      961 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     1021 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     1081 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     1141 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     1201 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     1261 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     1321 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     1381 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     1441 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     1501 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     1561 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     1621 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     1681 atctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     1741 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     1801 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     1861 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     1921 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     1981 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2041 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2101 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     2161 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2221 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2281 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2341 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2401 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2461 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2521 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2581 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2641 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2701 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2761 ggaagagcgc
//
