LOCUS       sequence-64876-e        5468 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 5468)
  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 5468)
  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: pXP711
FEATURES             Location/Qualifiers
     source          1..5468
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      734..981
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(750..768)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     protein_bind    1027..1060
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2502..2535
                     /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(3065..3081)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3065..3081)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3078..3100)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3089..3105
                     /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(3113..3119,3120..3137,3138..3143))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3158..3179
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3296..3313)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3467..4055)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3547..3566)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4226..5017,5018..5086))
                     /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     4849..4868
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5087..5191)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5259..5277
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5315..5337)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5437..5456
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catatgaggc atttgcaaga attactcgtg agtaaggaaa gagtgaggaa ctatcgcata
       61 cctgcattta aagatgccga tttgggcgcg aatcctttat tttggcttca ccctcatact
      121 attatcaggg ccagaaaaag gaagtgtttc cctccttctt gaattgatgt taccctcata
      181 aagcacgtgg cctcttatcg agaaagaaat taccgtcgct cgtgatttgt ttgcaaaaag
      241 aacaaaactg aaaaaaccca gacacgctcg acttcctgtc ttcctattga ttgcagcttc
      301 caatttcgtc acacaacaag gtcctagcga cggctcacag gttttgtaac aagcaatcga
      361 aggttctgga atggcgggaa agggtttagt accacatgct atgatgccca ctgtgatctc
      421 cagagcaaag ttcgttcgat cgtactgtta ctctctctct ttcaaacaga attgtccgaa
      481 tcgtgtgaca acaacagcct gttctcacac actcttttct tctaaccaag ggggtggttt
      541 agtttagtag aacctcgtga aacttacatt tacatatata taaacttgca taaattggtc
      601 aatgcaagaa atacatattt ggtcttttct aattcgtagt ttttcaagtt cttagatgct
      661 ttctttttct cttttttaca gatcatcaag gaagtaatta tctacttttt acaacaaata
      721 tactagtctc gagtcatgta attagttatg tcacgcttac attcacgccc tccccccaca
      781 tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt
      841 tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
      901 tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga
      961 cgctcgaagg ctttaatttg cggccaagct tgcatgcctg caggtcgact ctagaggatc
     1021 cccgggataa cttcgtataa tgtatgctat acgaagttat ggattttctt aacttcttcg
     1081 gcgacagcat caccgacttc ggtggtactg ttggaaccac ctaaatcacc agttctgata
     1141 cctgcatcca aaaccttttt aactgcatct tcaatggcct taccttcttc aggcaagttc
     1201 aatgacaatt tcaacatcat tgcagcagac aagatagtgg cgatagggtt gaccttattc
     1261 tttggcaaat ctggagcaga accgtggcat ggttcgtaca aaccaaatgc ggtgttcttg
     1321 tctggcaaag aggccaagga cgcagatggc aacaaaccca aggaacctgg gataacggag
     1381 gcttcatcgg agatgatatc accaaacatg ttgctggtga ttataatacc atttaggtgg
     1441 gttgggttct taactaggat catggcggca gaatcaatca attgatgttg aaccttcaat
     1501 gtaggaaatt cgttcttgat ggtttcctcc acagtttttc tccataatct tgaagaggcc
     1561 aaaacattag ctttatccaa ggaccaaata ggcaatggtg gctcatgttg tagggccatg
     1621 aaagcggcca ttcttgtgat tctttgcact tctggaacgg tgtattgttc actatcccaa
     1681 gcgacaccat caccatcatc ttcctttctc ttaccaaagt aaatacctcc cactaattct
     1741 ctgacaacaa cgaagtcagt acctttagca aattgtggct tgattggaga taagtctaaa
     1801 agagagtcgg atgcaaagtt acatggtctt aagttggcgt acaattgaag ttctttacgg
     1861 atttttagta aaccttgttc aggtctaaca ctacctgtac cccatttagg accacccaca
     1921 gcacctaaca aaacggcatc agccttcttg gaggcttcca gcgcctcatc tggaagtggg
     1981 acacctgtag cttcgatagc agcaccacca attaaatgat tttcgaaatc gaacttgaca
     2041 ttggaacgaa catcagaaat agctttaaga accttaatgg cttcggctgt gatttcttga
     2101 ccaacgtggt cacctggcaa aacgacgatc ttcttagggg cagacattag aatggtatat
     2161 ccttgaaata tatatatata ttgctgaaat gtaaaaggta agaaaagtta gaaagtaaga
     2221 cgattgctaa ccacctattg gaaaaacaat aggtccttaa ataatattgt caacttcaag
     2281 tattgtgatg caagcattta gtcatgaacg cttctctatt ctatatgaaa agccggttcc
     2341 ggcgctctca cctttccttt ttctcccaat ttttcagttg aaaaaggtat atgcgtcagg
     2401 cgacctctga aattaacaaa aaatttccag tcatcgaatt tgattctgtg cgatagcgcc
     2461 cctgtgtgtt ctcgttatgt tgaggaaaaa aataatggtt aataacttcg tataatgtat
     2521 gctatacgaa gttatcccgg gtaccgagct cgaattcatc acgtgctata aaaataatta
     2581 taatttaaat tttttaatat aaatatataa attaaaaata gaaagtaaaa aaagaaatta
     2641 aagaaaaaat agtttttgtt ttccgaagat gtaaaagact ctagggggat cgccaacaaa
     2701 tactaccttt tatcttgctc ttcctgctct caggtattaa tgccgaattg tttcatcttg
     2761 tctgtgtaga agaccacaca cgaaaatcct gtgattttac attttactta tcgttaatcg
     2821 aatgtatatc tatttaatct gcttttcttg tctaataaat atatatgtaa agtacgcttt
     2881 ttgttgaaat tttttaaacc tttgtttatt tttttttctt cattccgtaa ctcttctacc
     2941 ttctttattt actttctaaa atccaaatac aaaacataaa aataaataaa cacagagtaa
     3001 attcccaaat tattccatca ttaaaagata cgaggcgcgt gtaagttacg aattcgtaat
     3061 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     3121 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     3181 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     3241 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     3301 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3361 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3421 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3481 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3541 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3601 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3661 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3721 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3781 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3841 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3901 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3961 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4021 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4081 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4141 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4201 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4261 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4321 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4381 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4441 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4501 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4561 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4621 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4681 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4741 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4801 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4861 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4921 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4981 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5041 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5101 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5161 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     5221 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     5281 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     5341 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     5401 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     5461 gagtgcac
//
