LOCUS       sequence-65293-e        5914 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 5914)
  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 5914)
  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: pXP831
FEATURES             Location/Qualifiers
     source          1..5914
                     /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)."
     rep_origin      2149..3491
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(3112..3159)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(3511..3527)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3511..3527)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3524..3546)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3535..3551
                     /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(3559..3565,3566..3583,3584..3589))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3604..3625
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3742..3759)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3913..4501)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3993..4012)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4672..5463,5464..5532))
                     /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     5295..5314
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5533..5637)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5705..5723
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5761..5783)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5883..5902
                     /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 tcgaattcaa cgaagcatct
     2161 gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat
     2221 ctgagctgca tttttacaga acagaaatgc aacgcgaaag cgctatttta ccaacgaaga
     2281 atctgtgctt catttttgta aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa
     2341 agaatctgag ctgcattttt acagaacaga aatgcaacgc gagagcgcta ttttaccaac
     2401 aaagaatcta tacttctttt ttgttctaca aaaatgcatc ccgagagcgc tatttttcta
     2461 acaaagcatc ttagattact ttttttctcc tttgtgcgct ctataatgca gtctcttgat
     2521 aactttttgc actgtaggtc cgttaaggtt agaagaaggc tactttggtg tctattttct
     2581 cttccataaa aaaagcctga ctccacttcc cgcgtttact gattactagc gaagctgcgg
     2641 gtgcattttt tcaagataaa ggcatccccg attatattct ataccgatgt ggattgcgca
     2701 tactttgtga acagaaagtg atagcgttga tgattcttca ttggtcagaa aattatgaac
     2761 ggtttcttct attttgtctc tatatactac gtataggaaa tgtttacatt ttcgtattgt
     2821 tttcgattca ctctatgaat agttcttact acaatttttt tgtctaaaga gtaatactag
     2881 agataaacat aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg
     2941 atgggtaggt tatataggga tatagcacag agatatatag caaagagata cttttgagca
     3001 atgtttgtgg aagcggtatt cgcaatattt tagtagctcg ttacagtccg gtgcgttttt
     3061 ggttttttga aagtgcgtct tcagagcgct tttggttttc aaaagcgctc tgaagttcct
     3121 atactttcta gagaatagga acttcggaat aggaacttca aagcgtttcc gaaaacgagc
     3181 gcttccgaaa atgcaacgcg agctgcgcac atacagctca ctgttcacgt cgcacctata
     3241 tctgcgtgtt gcctgtatat atatatacat gagaagaacg gcatagtgcg tgtttatgct
     3301 taaatgcgta cttatatgcg tctatttatg taggatgaaa ggtagtctag tacctcctgt
     3361 gatattatcc cattccatgc ggggtatcgt atgcttcctt cagcactacc ctttagctgt
     3421 tctatatgct gccactcctc aattggatta gtctcatcct tcaatgctat catttccttt
     3481 gatattggat catacgaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3541 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3601 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3661 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3721 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3781 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3841 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3901 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3961 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4021 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4081 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4141 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4201 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4261 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4321 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     4381 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4441 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4501 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4561 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4621 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4681 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4741 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4801 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4861 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4921 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4981 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5041 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5101 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5161 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5221 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5281 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5341 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5401 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5461 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5521 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5581 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5641 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     5701 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     5761 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     5821 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     5881 tgcggcatca gagcagattg tactgagagt gcac
//
