LOCUS       sequence-12714-e        4821 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 4821)
  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 4821)
  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: pXP518
FEATURES             Location/Qualifiers
     source          1..4821
                     /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)."
     primer_bind     complement(2600..2616)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2600..2616)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2613..2635)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2624..2640
                     /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(2648..2654,2655..2672,2673..2678))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2693..2714
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2831..2848)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3002..3590)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3082..3101)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3761..4552,4553..4621))
                     /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     4384..4403
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4622..4726)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4794..4812
                     /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 ttcgtaactt
     2101 acacgcgcct cgtatctttt aatgatggaa taatttggga atttactctg tgtttattta
     2161 tttttatgtt ttgtatttgg attttagaaa gtaaataaag aaggtagaag agttacggaa
     2221 tgaagaaaaa aaaataaaca aaggtttaaa aaatttcaac aaaaagcgta ctttacatat
     2281 atatttatta gacaagaaaa gcagattaaa tagatataca ttcgattaac gataagtaaa
     2341 atgtaaaatc acaggatttt cgtgtgtggt cttctacaca gacaagatga aacaattcgg
     2401 cattaatacc tgagagcagg aagagcaaga taaaaggtag tatttgttgg cgatccccct
     2461 agagtctttt acatcttcgg aaaacaaaaa ctattttttc tttaatttct ttttttactt
     2521 tctattttta atttatatat ttatattaaa aaatttaaat tataattatt tttatagcac
     2581 gtgatgaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     2641 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     2701 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     2761 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2821 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2881 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2941 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3001 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3061 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3121 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3181 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3241 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3301 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3361 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3421 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     3481 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3541 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3601 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3661 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3721 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3781 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3841 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3901 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3961 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4021 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4081 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4141 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4201 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4261 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4321 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4381 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4441 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4501 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4561 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4621 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4681 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4741 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     4801 gtatcacgag gccctttcgt c
//
