LOCUS       sequence-65289-e        6323 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 6323)
  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 6323)
  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: pXP811
FEATURES             Location/Qualifiers
     source          1..6323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      734..981
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(750..768)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     protein_bind    1027..1060
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2502..2535
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      2558..3900
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(3521..3568)
                     /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(3920..3936)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3920..3936)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3933..3955)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3944..3960
                     /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(3968..3974,3975..3992,3993..3998))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4013..4034
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4151..4168)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4322..4910)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4402..4421)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5081..5872,5873..5941))
                     /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     5704..5723
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5942..6046)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6114..6132
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6170..6192)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6292..6311
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catatgaggc atttgcaaga attactcgtg agtaaggaaa gagtgaggaa ctatcgcata
       61 cctgcattta aagatgccga tttgggcgcg aatcctttat tttggcttca ccctcatact
      121 attatcaggg ccagaaaaag gaagtgtttc cctccttctt gaattgatgt taccctcata
      181 aagcacgtgg cctcttatcg agaaagaaat taccgtcgct cgtgatttgt ttgcaaaaag
      241 aacaaaactg aaaaaaccca gacacgctcg acttcctgtc ttcctattga ttgcagcttc
      301 caatttcgtc acacaacaag gtcctagcga cggctcacag gttttgtaac aagcaatcga
      361 aggttctgga atggcgggaa agggtttagt accacatgct atgatgccca ctgtgatctc
      421 cagagcaaag ttcgttcgat cgtactgtta ctctctctct ttcaaacaga attgtccgaa
      481 tcgtgtgaca acaacagcct gttctcacac actcttttct tctaaccaag ggggtggttt
      541 agtttagtag aacctcgtga aacttacatt tacatatata taaacttgca taaattggtc
      601 aatgcaagaa atacatattt ggtcttttct aattcgtagt ttttcaagtt cttagatgct
      661 ttctttttct cttttttaca gatcatcaag gaagtaatta tctacttttt acaacaaata
      721 tactagtctc gagtcatgta attagttatg tcacgcttac attcacgccc tccccccaca
      781 tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt
      841 tttatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
      901 tacagacgcg tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga
      961 cgctcgaagg ctttaatttg cggccaagct tgcatgcctg caggtcgact ctagaggatc
     1021 cccgggataa cttcgtataa tgtatgctat acgaagttat ggattttctt aacttcttcg
     1081 gcgacagcat caccgacttc ggtggtactg ttggaaccac ctaaatcacc agttctgata
     1141 cctgcatcca aaaccttttt aactgcatct tcaatggcct taccttcttc aggcaagttc
     1201 aatgacaatt tcaacatcat tgcagcagac aagatagtgg cgatagggtt gaccttattc
     1261 tttggcaaat ctggagcaga accgtggcat ggttcgtaca aaccaaatgc ggtgttcttg
     1321 tctggcaaag aggccaagga cgcagatggc aacaaaccca aggaacctgg gataacggag
     1381 gcttcatcgg agatgatatc accaaacatg ttgctggtga ttataatacc atttaggtgg
     1441 gttgggttct taactaggat catggcggca gaatcaatca attgatgttg aaccttcaat
     1501 gtaggaaatt cgttcttgat ggtttcctcc acagtttttc tccataatct tgaagaggcc
     1561 aaaacattag ctttatccaa ggaccaaata ggcaatggtg gctcatgttg tagggccatg
     1621 aaagcggcca ttcttgtgat tctttgcact tctggaacgg tgtattgttc actatcccaa
     1681 gcgacaccat caccatcatc ttcctttctc ttaccaaagt aaatacctcc cactaattct
     1741 ctgacaacaa cgaagtcagt acctttagca aattgtggct tgattggaga taagtctaaa
     1801 agagagtcgg atgcaaagtt acatggtctt aagttggcgt acaattgaag ttctttacgg
     1861 atttttagta aaccttgttc aggtctaaca ctacctgtac cccatttagg accacccaca
     1921 gcacctaaca aaacggcatc agccttcttg gaggcttcca gcgcctcatc tggaagtggg
     1981 acacctgtag cttcgatagc agcaccacca attaaatgat tttcgaaatc gaacttgaca
     2041 ttggaacgaa catcagaaat agctttaaga accttaatgg cttcggctgt gatttcttga
     2101 ccaacgtggt cacctggcaa aacgacgatc ttcttagggg cagacattag aatggtatat
     2161 ccttgaaata tatatatata ttgctgaaat gtaaaaggta agaaaagtta gaaagtaaga
     2221 cgattgctaa ccacctattg gaaaaacaat aggtccttaa ataatattgt caacttcaag
     2281 tattgtgatg caagcattta gtcatgaacg cttctctatt ctatatgaaa agccggttcc
     2341 ggcgctctca cctttccttt ttctcccaat ttttcagttg aaaaaggtat atgcgtcagg
     2401 cgacctctga aattaacaaa aaatttccag tcatcgaatt tgattctgtg cgatagcgcc
     2461 cctgtgtgtt ctcgttatgt tgaggaaaaa aataatggtt aataacttcg tataatgtat
     2521 gctatacgaa gttatcccgg gtaccgagct cgaattcaac gaagcatctg tgcttcattt
     2581 tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
     2641 ttttacagaa cagaaatgca acgcgaaagc gctattttac caacgaagaa tctgtgcttc
     2701 atttttgtaa aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc
     2761 tgcattttta cagaacagaa atgcaacgcg agagcgctat tttaccaaca aagaatctat
     2821 acttcttttt tgttctacaa aaatgcatcc cgagagcgct atttttctaa caaagcatct
     2881 tagattactt tttttctcct ttgtgcgctc tataatgcag tctcttgata actttttgca
     2941 ctgtaggtcc gttaaggtta gaagaaggct actttggtgt ctattttctc ttccataaaa
     3001 aaagcctgac tccacttccc gcgtttactg attactagcg aagctgcggg tgcatttttt
     3061 caagataaag gcatccccga ttatattcta taccgatgtg gattgcgcat actttgtgaa
     3121 cagaaagtga tagcgttgat gattcttcat tggtcagaaa attatgaacg gtttcttcta
     3181 ttttgtctct atatactacg tataggaaat gtttacattt tcgtattgtt ttcgattcac
     3241 tctatgaata gttcttacta caattttttt gtctaaagag taatactaga gataaacata
     3301 aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc gaaaggtgga tgggtaggtt
     3361 atatagggat atagcacaga gatatatagc aaagagatac ttttgagcaa tgtttgtgga
     3421 agcggtattc gcaatatttt agtagctcgt tacagtccgg tgcgtttttg gttttttgaa
     3481 agtgcgtctt cagagcgctt ttggttttca aaagcgctct gaagttccta tactttctag
     3541 agaataggaa cttcggaata ggaacttcaa agcgtttccg aaaacgagcg cttccgaaaa
     3601 tgcaacgcga gctgcgcaca tacagctcac tgttcacgtc gcacctatat ctgcgtgttg
     3661 cctgtatata tatatacatg agaagaacgg catagtgcgt gtttatgctt aaatgcgtac
     3721 ttatatgcgt ctatttatgt aggatgaaag gtagtctagt acctcctgtg atattatccc
     3781 attccatgcg gggtatcgta tgcttccttc agcactaccc tttagctgtt ctatatgctg
     3841 ccactcctca attggattag tctcatcctt caatgctatc atttcctttg atattggatc
     3901 atacgaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     3961 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4021 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4081 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4141 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4201 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4261 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4321 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4381 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4441 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4501 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4561 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4621 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4681 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4741 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     4801 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4861 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4921 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4981 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5041 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5101 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5161 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5221 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5281 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5341 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5401 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5461 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5521 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5581 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5641 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5701 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5761 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5821 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5881 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5941 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6001 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6061 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     6121 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     6181 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     6241 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     6301 agcagattgt actgagagtg cac
//
