LOCUS       sequence-65291-e        6060 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 6060)
  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 6060)
  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: pXP821
FEATURES             Location/Qualifiers
     source          1..6060
                     /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)."
     rep_origin      2295..3637
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(3258..3305)
                     /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(3657..3673)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3657..3673)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3670..3692)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3681..3697
                     /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(3705..3711,3712..3729,3730..3735))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3750..3771
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3888..3905)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4059..4647)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4139..4158)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4818..5609,5610..5678))
                     /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     5441..5460
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5679..5783)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5851..5869
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5907..5929)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6029..6048
                     /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 attcaacgaa gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga
     2341 gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag aaatgcaacg
     2401 cgaaagcgct attttaccaa cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa
     2461 cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg
     2521 caacgcgaga gcgctatttt accaacaaag aatctatact tcttttttgt tctacaaaaa
     2581 tgcatcccga gagcgctatt tttctaacaa agcatcttag attacttttt ttctcctttg
     2641 tgcgctctat aatgcagtct cttgataact ttttgcactg taggtccgtt aaggttagaa
     2701 gaaggctact ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc acttcccgcg
     2761 tttactgatt actagcgaag ctgcgggtgc attttttcaa gataaaggca tccccgatta
     2821 tattctatac cgatgtggat tgcgcatact ttgtgaacag aaagtgatag cgttgatgat
     2881 tcttcattgg tcagaaaatt atgaacggtt tcttctattt tgtctctata tactacgtat
     2941 aggaaatgtt tacattttcg tattgttttc gattcactct atgaatagtt cttactacaa
     3001 tttttttgtc taaagagtaa tactagagat aaacataaaa aatgtagagg tcgagtttag
     3061 atgcaagttc aaggagcgaa aggtggatgg gtaggttata tagggatata gcacagagat
     3121 atatagcaaa gagatacttt tgagcaatgt ttgtggaagc ggtattcgca atattttagt
     3181 agctcgttac agtccggtgc gtttttggtt ttttgaaagt gcgtcttcag agcgcttttg
     3241 gttttcaaaa gcgctctgaa gttcctatac tttctagaga ataggaactt cggaatagga
     3301 acttcaaagc gtttccgaaa acgagcgctt ccgaaaatgc aacgcgagct gcgcacatac
     3361 agctcactgt tcacgtcgca cctatatctg cgtgttgcct gtatatatat atacatgaga
     3421 agaacggcat agtgcgtgtt tatgcttaaa tgcgtactta tatgcgtcta tttatgtagg
     3481 atgaaaggta gtctagtacc tcctgtgata ttatcccatt ccatgcgggg tatcgtatgc
     3541 ttccttcagc actacccttt agctgttcta tatgctgcca ctcctcaatt ggattagtct
     3601 catccttcaa tgctatcatt tcctttgata ttggatcata cgaattcgta atcatggtca
     3661 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     3721 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     3781 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     3841 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3901 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3961 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4021 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4081 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4141 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4201 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4261 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4321 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4381 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4441 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     4501 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4561 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4621 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4681 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4741 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4801 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4861 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4921 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4981 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5041 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5101 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5161 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5221 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5281 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5341 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5401 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5461 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5521 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5581 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5641 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5701 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5761 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     5821 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     5881 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     5941 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     6001 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
//
