LOCUS       sequence-12716-e        5677 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 5677)
  AUTHORS   Fang F, Salmon K, Shen MW, Aeling KA, Ito E, Irwin B, Tran UP, 
            Hatfield GW, Da Silva NA, Sandmeyer S
  TITLE     A vector set for systematic metabolic engineering in Saccharomyces 
            cerevisiae.
  JOURNAL   Yeast. 2011 Feb;28(2):123-36. doi: 10.1002/yea.1824. Epub 2010 Oct 
            8.
  PUBMED    20936606
REFERENCE   2  (bases 1 to 5677)
  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.1824"; 
            journalName: "Yeast"; date: "2011-02"; volume: "28"; issue: "2"; 
            pages: "123-36"
COMMENT     Sequence Label: pXP618
FEATURES             Location/Qualifiers
     source          1..5677
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      598..845
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(614..632)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     protein_bind    complement(891..924)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        940..1155
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             1156..1959
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     protein_bind    complement(2038..2071)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      2094..3436
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(3057..3104)
                     /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(3456..3472)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3456..3472)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3469..3491)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3480..3496
                     /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(3504..3510,3511..3528,3529..3534))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3549..3570
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3687..3704)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3858..4446)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3938..3957)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4617..5408,5409..5477))
                     /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     5240..5259
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5478..5582)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5650..5668
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgct cgtaggaaca atttcgggcc cctgcgtgtt cttctgaggt tcatctttta
      241 catttgcttc tgctggataa ttttcagagg caacaaggaa aaattagatg gcaaaaagtc
      301 gtctttcaag gaaaaatccc caccatcttt cgagatcccc tgtaacttat tggcaactga
      361 aagaatgaaa aggaggaaaa tacaaaatat actagaactg aaaaaaaaaa agtataaata
      421 gagacgatat atgccaatac ttcacaatgt tcgaatctat tcttcatttg cagctattgt
      481 aaaataataa aacatcaaga acaaacaagc tcaacttgtc ttttctaaga acaaagaata
      541 aacacaaaaa caaaaagttt ttttaatttt aatcaaaaaa ctagtgatat cctcgagtca
      601 tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct ctaaccgaaa
      661 aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata gttatgttag
      721 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg
      781 catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg aaggctttaa
      841 tttgcggcca agcttgcatg cctgcaggtc gactctagag gatccccggg ataacttcgt
      901 atagcataca ttatacgaag ttatgcacca taccaccttt tcaattcatc attttttttt
      961 tattcttttt tttgatttcg gtttccttga aatttttttg attcggtaat ctccgaacag
     1021 aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt
     1081 gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa aacctgcagg
     1141 aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt
     1201 cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca
     1261 ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt
     1321 tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag
     1381 ccgctaaagg cattatccgc caagtacaat tttttactct tcgaagacag aaaatttgct
     1441 gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg
     1501 gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag
     1561 gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc
     1621 aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac
     1681 aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac
     1741 gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagacgc attgggtcaa
     1801 cagtatagaa ccgtggatga tgtggtctct acaggatctg acattattat tgttggaaga
     1861 ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc
     1921 tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat
     1981 gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt attacccata
     2041 acttcgtata gcatacatta tacgaagtta tcccgggtac cgagctcgaa ttcaacgaag
     2101 catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa
     2161 agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta ttttaccaac
     2221 gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca
     2281 aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag cgctatttta
     2341 ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag agcgctattt
     2401 ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata atgcagtctc
     2461 ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt tggtgtctat
     2521 tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta ctagcgaagc
     2581 tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc gatgtggatt
     2641 gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt cagaaaatta
     2701 tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt acattttcgt
     2761 attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct aaagagtaat
     2821 actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca aggagcgaaa
     2881 ggtggatggg taggttatat agggatatag cacagagata tatagcaaag agatactttt
     2941 gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca gtccggtgcg
     3001 tttttggttt tttgaaagtg cgtcttcaga gcgcttttgg ttttcaaaag cgctctgaag
     3061 ttcctatact ttctagagaa taggaacttc ggaataggaa cttcaaagcg tttccgaaaa
     3121 cgagcgcttc cgaaaatgca acgcgagctg cgcacataca gctcactgtt cacgtcgcac
     3181 ctatatctgc gtgttgcctg tatatatata tacatgagaa gaacggcata gtgcgtgttt
     3241 atgcttaaat gcgtacttat atgcgtctat ttatgtagga tgaaaggtag tctagtacct
     3301 cctgtgatat tatcccattc catgcggggt atcgtatgct tccttcagca ctacccttta
     3361 gctgttctat atgctgccac tcctcaattg gattagtctc atccttcaat gctatcattt
     3421 cctttgatat tggatcatac gaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     3481 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     3541 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     3601 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3661 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3721 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3781 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3841 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3901 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3961 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4021 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4081 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4141 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4201 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4261 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     4321 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4381 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4441 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4501 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4561 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4621 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4681 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4741 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4801 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4861 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4921 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4981 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5041 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5101 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5161 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5221 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5281 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5341 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5401 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5461 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5521 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5581 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     5641 acctataaaa ataggcgtat cacgaggccc tttcgtc
//
