LOCUS       sequence-64879-e        5059 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5059)
  AUTHORS   Shen MW, Fang F, Sandmeyer S, Da Silva NA
  TITLE     Development and characterization of a vector set with regulated 
            promoters for systematic metabolic engineering in Saccharomyces 
            cerevisiae.
  JOURNAL   Yeast. 2012 Dec;29(12):495-503. doi: 10.1002/yea.2930. Epub 2012 Nov
            19.
  PUBMED    23166051
REFERENCE   2  (bases 1 to 5059)
  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"; doi: "10.1002/yea.2930"; 
            journalName: "Yeast"; date: "2012-12"; volume: "29"; issue: "12"; 
            pages: "495-503"
COMMENT     Sequence Label: pXP731
FEATURES             Location/Qualifiers
     source          1..5059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..256
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     terminator      325..572
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(341..359)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     protein_bind    618..651
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2093..2126
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(2656..2672)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2656..2672)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2669..2691)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2680..2696
                     /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(2704..2710,2711..2728,2729..2734))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2749..2770
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2887..2904)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3058..3646)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3138..3157)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3817..4608,4609..4677))
                     /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     4440..4459
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4678..4782)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4850..4868
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4906..4928)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5028..5047
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catatgatct tagccttgtt actagttaga aaaagacatt tttgctgtca gtcactgtca
       61 agagattctt ttgctggcat ttcttctaga agcaaaaaga gcgatgcgtc ttttccgctg
      121 aaccgttcca gcaaaaaaga ctaccaacgc aatatggatt gtcagaatca tataaaagag
      181 aagcaaataa ctccttgtct tgtatcaatt gcattataat atcttcttgt tagtgcaata
      241 tcatatagaa gtcatcgaaa tagatattaa gaaaaacaaa ctgtacaatc aatcaatcaa
      301 tcatcacata aacctaggct cgagtcatgt aattagttat gtcacgctta cattcacgcc
      361 ctccccccac atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc
      421 ctatttattt ttttatagtt atgttagtat taagaacgtt atttatattt caaatttttc
      481 ttttttttct gtacagacgc gtgtacgcat gtaacattat actgaaaacc ttgcttgaga
      541 aggttttggg acgctcgaag gctttaattt gcggccaagc ttgcatgcct gcaggtcgac
      601 tctagaggat ccccgggata acttcgtata atgtatgcta tacgaagtta tggattttct
      661 taacttcttc ggcgacagca tcaccgactt cggtggtact gttggaacca cctaaatcac
      721 cagttctgat acctgcatcc aaaacctttt taactgcatc ttcaatggcc ttaccttctt
      781 caggcaagtt caatgacaat ttcaacatca ttgcagcaga caagatagtg gcgatagggt
      841 tgaccttatt ctttggcaaa tctggagcag aaccgtggca tggttcgtac aaaccaaatg
      901 cggtgttctt gtctggcaaa gaggccaagg acgcagatgg caacaaaccc aaggaacctg
      961 ggataacgga ggcttcatcg gagatgatat caccaaacat gttgctggtg attataatac
     1021 catttaggtg ggttgggttc ttaactagga tcatggcggc agaatcaatc aattgatgtt
     1081 gaaccttcaa tgtaggaaat tcgttcttga tggtttcctc cacagttttt ctccataatc
     1141 ttgaagaggc caaaacatta gctttatcca aggaccaaat aggcaatggt ggctcatgtt
     1201 gtagggccat gaaagcggcc attcttgtga ttctttgcac ttctggaacg gtgtattgtt
     1261 cactatccca agcgacacca tcaccatcat cttcctttct cttaccaaag taaatacctc
     1321 ccactaattc tctgacaaca acgaagtcag tacctttagc aaattgtggc ttgattggag
     1381 ataagtctaa aagagagtcg gatgcaaagt tacatggtct taagttggcg tacaattgaa
     1441 gttctttacg gatttttagt aaaccttgtt caggtctaac actacctgta ccccatttag
     1501 gaccacccac agcacctaac aaaacggcat cagccttctt ggaggcttcc agcgcctcat
     1561 ctggaagtgg gacacctgta gcttcgatag cagcaccacc aattaaatga ttttcgaaat
     1621 cgaacttgac attggaacga acatcagaaa tagctttaag aaccttaatg gcttcggctg
     1681 tgatttcttg accaacgtgg tcacctggca aaacgacgat cttcttaggg gcagacatta
     1741 gaatggtata tccttgaaat atatatatat attgctgaaa tgtaaaaggt aagaaaagtt
     1801 agaaagtaag acgattgcta accacctatt ggaaaaacaa taggtcctta aataatattg
     1861 tcaacttcaa gtattgtgat gcaagcattt agtcatgaac gcttctctat tctatatgaa
     1921 aagccggttc cggcgctctc acctttcctt tttctcccaa tttttcagtt gaaaaaggta
     1981 tatgcgtcag gcgacctctg aaattaacaa aaaatttcca gtcatcgaat ttgattctgt
     2041 gcgatagcgc ccctgtgtgt tctcgttatg ttgaggaaaa aaataatggt taataacttc
     2101 gtataatgta tgctatacga agttatcccg ggtaccgagc tcgaattcat cacgtgctat
     2161 aaaaataatt ataatttaaa ttttttaata taaatatata aattaaaaat agaaagtaaa
     2221 aaaagaaatt aaagaaaaaa tagtttttgt tttccgaaga tgtaaaagac tctaggggga
     2281 tcgccaacaa atactacctt ttatcttgct cttcctgctc tcaggtatta atgccgaatt
     2341 gtttcatctt gtctgtgtag aagaccacac acgaaaatcc tgtgatttta cattttactt
     2401 atcgttaatc gaatgtatat ctatttaatc tgcttttctt gtctaataaa tatatatgta
     2461 aagtacgctt tttgttgaaa ttttttaaac ctttgtttat ttttttttct tcattccgta
     2521 actcttctac cttctttatt tactttctaa aatccaaata caaaacataa aaataaataa
     2581 acacagagta aattcccaaa ttattccatc attaaaagat acgaggcgcg tgtaagttac
     2641 gaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     2701 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     2761 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     2821 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2881 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2941 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3001 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3061 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3121 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3181 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3241 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3301 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3361 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3421 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3481 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3541 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3601 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3661 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3721 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3781 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3841 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3901 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3961 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4021 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4081 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4141 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4201 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     4261 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4321 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4381 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4441 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4501 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4561 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4621 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4681 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4741 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4801 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     4861 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     4921 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     4981 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     5041 gattgtactg agagtgcac
//
