LOCUS       sequence-64878-e        5205 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 5205)
  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 5205)
  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: pXP721
FEATURES             Location/Qualifiers
     source          1..5205
                     /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    764..797
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2239..2272
                     /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(2802..2818)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2802..2818)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2815..2837)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2826..2842
                     /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(2850..2856,2857..2874,2875..2880))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2895..2916
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3033..3050)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3204..3792)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3284..3303)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3963..4754,4755..4823))
                     /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     4586..4605
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4824..4928)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4996..5014
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5052..5074)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5174..5193
                     /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 cgtataatgt
      781 atgctatacg aagttatgga ttttcttaac ttcttcggcg acagcatcac cgacttcggt
      841 ggtactgttg gaaccaccta aatcaccagt tctgatacct gcatccaaaa cctttttaac
      901 tgcatcttca atggccttac cttcttcagg caagttcaat gacaatttca acatcattgc
      961 agcagacaag atagtggcga tagggttgac cttattcttt ggcaaatctg gagcagaacc
     1021 gtggcatggt tcgtacaaac caaatgcggt gttcttgtct ggcaaagagg ccaaggacgc
     1081 agatggcaac aaacccaagg aacctgggat aacggaggct tcatcggaga tgatatcacc
     1141 aaacatgttg ctggtgatta taataccatt taggtgggtt gggttcttaa ctaggatcat
     1201 ggcggcagaa tcaatcaatt gatgttgaac cttcaatgta ggaaattcgt tcttgatggt
     1261 ttcctccaca gtttttctcc ataatcttga agaggccaaa acattagctt tatccaagga
     1321 ccaaataggc aatggtggct catgttgtag ggccatgaaa gcggccattc ttgtgattct
     1381 ttgcacttct ggaacggtgt attgttcact atcccaagcg acaccatcac catcatcttc
     1441 ctttctctta ccaaagtaaa tacctcccac taattctctg acaacaacga agtcagtacc
     1501 tttagcaaat tgtggcttga ttggagataa gtctaaaaga gagtcggatg caaagttaca
     1561 tggtcttaag ttggcgtaca attgaagttc tttacggatt tttagtaaac cttgttcagg
     1621 tctaacacta cctgtacccc atttaggacc acccacagca cctaacaaaa cggcatcagc
     1681 cttcttggag gcttccagcg cctcatctgg aagtgggaca cctgtagctt cgatagcagc
     1741 accaccaatt aaatgatttt cgaaatcgaa cttgacattg gaacgaacat cagaaatagc
     1801 tttaagaacc ttaatggctt cggctgtgat ttcttgacca acgtggtcac ctggcaaaac
     1861 gacgatcttc ttaggggcag acattagaat ggtatatcct tgaaatatat atatatattg
     1921 ctgaaatgta aaaggtaaga aaagttagaa agtaagacga ttgctaacca cctattggaa
     1981 aaacaatagg tccttaaata atattgtcaa cttcaagtat tgtgatgcaa gcatttagtc
     2041 atgaacgctt ctctattcta tatgaaaagc cggttccggc gctctcacct ttcctttttc
     2101 tcccaatttt tcagttgaaa aaggtatatg cgtcaggcga cctctgaaat taacaaaaaa
     2161 tttccagtca tcgaatttga ttctgtgcga tagcgcccct gtgtgttctc gttatgttga
     2221 ggaaaaaaat aatggttaat aacttcgtat aatgtatgct atacgaagtt atcccgggta
     2281 ccgagctcga attcatcacg tgctataaaa ataattataa tttaaatttt ttaatataaa
     2341 tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag aaaaaatagt ttttgttttc
     2401 cgaagatgta aaagactcta gggggatcgc caacaaatac taccttttat cttgctcttc
     2461 ctgctctcag gtattaatgc cgaattgttt catcttgtct gtgtagaaga ccacacacga
     2521 aaatcctgtg attttacatt ttacttatcg ttaatcgaat gtatatctat ttaatctgct
     2581 tttcttgtct aataaatata tatgtaaagt acgctttttg ttgaaatttt ttaaaccttt
     2641 gtttattttt ttttcttcat tccgtaactc ttctaccttc tttatttact ttctaaaatc
     2701 caaatacaaa acataaaaat aaataaacac agagtaaatt cccaaattat tccatcatta
     2761 aaagatacga ggcgcgtgta agttacgaat tcgtaatcat ggtcatagct gtttcctgtg
     2821 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     2881 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     2941 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3001 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3061 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3121 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3181 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3241 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3301 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3361 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3421 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3481 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3541 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3601 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     3661 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     3721 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     3781 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     3841 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     3901 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     3961 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4021 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4081 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4141 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4201 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4261 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4321 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4381 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4441 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4501 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4561 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4621 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4681 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     4741 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     4801 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     4861 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     4921 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     4981 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     5041 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     5101 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     5161 tggcttaact atgcggcatc agagcagatt gtactgagag tgcac
//
