LOCUS       sequence-64881-e        4880 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 4880)
  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 4880)
  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: pXP722
FEATURES             Location/Qualifiers
     source          1..4880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..450
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    8..125
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     421..444
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     terminator      471..718
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(487..505)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     protein_bind    complement(764..797)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        815..1031
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             1032..1835
                     /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
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLATGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     protein_bind    complement(1914..1947)
                     /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(2477..2493)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2477..2493)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2490..2512)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2501..2517
                     /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(2525..2531,2532..2549,2550..2555))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2570..2591
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2708..2725)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2879..3467)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2959..2978)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3638..4429,4430..4498))
                     /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     4261..4280
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4499..4603)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4671..4689
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4727..4749)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4849..4868
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catatgacgg attagaagcc gccgagcggg tgacagccct ccgaaggaag actctcctcc
       61 gtgcgtcctc gtcctcaccg gtcgcgttcc tgaaacgcag atgtgcctcg cgccgcactg
      121 ctccgaacaa taaagattct acaatactag cttttatggt tatgaagagg aaaaattggc
      181 agtaacctgg ccccacaaac cttcaaatga acgaatcaaa ttaacaacca taggatgata
      241 atgcgattag ttttttagcc ttatttctgg ggtaattaat cagcgaagcg atgatttttg
      301 atctattaac agatatataa atgcaaaaac tgcattaacc actttaacta atactttcaa
      361 cattttcggt ttgtattact tcttattcaa atgtaataaa agtatcaaca aaaaattgtt
      421 aatatacctc tatactttaa cgtcaaggag aaaaaaccac tagtctcgag tcatgtaatt
      481 agttatgtca cgcttacatt cacgccctcc ccccacatcc gctctaaccg aaaaggaagg
      541 agttagacaa cctgaagtct aggtccctat ttattttttt atagttatgt tagtattaag
      601 aacgttattt atatttcaaa tttttctttt ttttctgtac agacgcgtgt acgcatgtaa
      661 cattatactg aaaaccttgc ttgagaaggt tttgggacgc tcgaaggctt taatttgcgg
      721 ccaagcttgc atgcctgcag gtcgactcta gaggatcccc gggataactt cgtatagcat
      781 acattatacg aagttatcag ggtccataaa gcttttcaat tcatcttttt tttttttgtt
      841 cttttttttg attccggttt ctttgaaatt tttttgattc ggtaatctcc gagcagaagg
      901 aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt ggtgttgaag
      961 aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacc tgcaggaaac
     1021 gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat cctagtcctg
     1081 ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt gcttcattgg
     1141 atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc aaaatttgtt
     1201 tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca gttaagccgc
     1261 taaaggcatt atccgccaag tacaattttt tactcttcga agacagaaaa tttgctgaca
     1321 ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca gaatgggcag
     1381 acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg aagcaggcgg
     1441 cggaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg tcatgcaagg
     1501 gctccctagc tactggagaa tatactaagg gtactgttga cattgcgaag agcgacaaag
     1561 attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa ggttacgatt
     1621 ggttgattat gacacccggt gtgggtttag atgacaaggg agacgcattg ggtcaacagt
     1681 atagaaccgt ggatgatgtg gtctctacag gatctgacat tattattgtt ggaagaggac
     1741 tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa gcaggctggg
     1801 aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa gtaaatgcat
     1861 gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta cccataactt
     1921 cgtatagcat acattatacg aagttatccc gggtaccgag ctcgaattca tcacgtgcta
     1981 taaaaataat tataatttaa attttttaat ataaatatat aaattaaaaa tagaaagtaa
     2041 aaaaagaaat taaagaaaaa atagtttttg ttttccgaag atgtaaaaga ctctaggggg
     2101 atcgccaaca aatactacct tttatcttgc tcttcctgct ctcaggtatt aatgccgaat
     2161 tgtttcatct tgtctgtgta gaagaccaca cacgaaaatc ctgtgatttt acattttact
     2221 tatcgttaat cgaatgtata tctatttaat ctgcttttct tgtctaataa atatatatgt
     2281 aaagtacgct ttttgttgaa attttttaaa cctttgttta tttttttttc ttcattccgt
     2341 aactcttcta ccttctttat ttactttcta aaatccaaat acaaaacata aaaataaata
     2401 aacacagagt aaattcccaa attattccat cattaaaaga tacgaggcgc gtgtaagtta
     2461 cgaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     2521 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     2581 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     2641 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2701 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2761 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2821 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2881 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2941 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3001 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3061 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3121 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3181 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3241 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3301 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3361 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3421 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3481 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3541 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3601 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3661 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3721 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3781 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3841 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3901 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3961 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4021 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4081 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4141 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4201 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4261 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4321 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4381 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4441 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4501 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4561 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     4621 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     4681 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     4741 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     4801 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     4861 agattgtact gagagtgcac
//
