LOCUS       sequence-320161-        8060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  (parent plasmid pCFB257) pX - 3 -LoxP -KlLEU2 -pTEF1 - TetR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8060)
  AUTHORS   Ambri F, D'Ambrosio V, Di Blasi R, Maury J, Jacobsen SAB, McCloskey 
            D, Jensen MK, Keasling JD
  TITLE     High-Resolution Scanning of Optimal Biosensor Reporter Promoters in 
            Yeast.
  JOURNAL   ACS Synth Biol. 2020 Feb 21;9(2):218-226. doi: 
            10.1021/acssynbio.9b00333. Epub 2020 Jan 31.
  PUBMED    31935067
REFERENCE   2  (bases 1 to 8060)
  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: 
            "10.1021/acssynbio.9b00333"; journalName: "ACS Synth Biol"; date: 
            "2020-02-21- 21"; volume: "9"; issue: "2"; pages: "218-226"
COMMENT     Sequence Label: pVD_79
FEATURES             Location/Qualifiers
     source          1..8060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(190..355)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     276..300
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     promoter        404..810
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     protein_bind    1714..1747
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4038..4071
                     /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(4114..4133)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4115..4133)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4143..4159)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4152..4174)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    complement(4822..4855)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(5325..5341)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5440..5457)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5611..6199)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5691..5710)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6321..7112,7113..7181))
                     /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     6944..6963
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7182..7273)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7315..7334
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7340..7359
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    7566..7599
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
ORIGIN
        1 tgcttggcct ttgttttggt ttttgatgtc ggtcgtttga ggccccttgc ggaaaatcga
       61 gatcgccgaa tggcacgcga gggaagggaa ataaggttta aaggcactga aacaataggc
      121 aagaagtagg cgagagccga catacgagac tattaagtcc tcagcgagct cgcatggaat
      181 gcgtgcgatg agcgacctca tgctatacct gagaaagcaa cctgacctac aggaaagagt
      241 tactcaagaa taagaatttt cgttttaaaa cctaagagtc actttaaaat ttgtatacac
      301 ttattttttt tataacttat ttaataataa aaatcataaa tcataagaaa ttcgcttatt
      361 tagaagtgtc aacaacgtat ctaccaacgg aatacgtgcg atgcacacac catagcttca
      421 aaatgtttct actccttttt tactcttcca gattttctcg gactccgcgc atcgccgtac
      481 cacttcaaaa cacccaagca cagcatacta aatttcccct ctttcttcct ctagggtgtc
      541 gttaattacc cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt ttctttttct
      601 tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt ttcttgaaaa tttttttttt
      661 tgattttttt ctctttcgat gacctcccat tgatatttaa gttaataaac ggtcttcaat
      721 ttctcaagtt tcagtttcat ttttcttgtt ctattacaac tttttttact tcttgctcat
      781 tagaaagaaa gcatagcaat ctaatctaag ttttaattac aaagtgcagg taaaacaatg
      841 atgtctagac tggacaaatc taaggttata aacagtgcac tagaattgtt gaatgaagtt
      901 ggtatcgagg gtcttactac gagaaagtta gcacaaaaat taggcgttga acaacctact
      961 ctttattggc atgtcaaaaa caagcgtgca ttgctggacg cactagctat agaaatgttg
     1021 gacagacacc acacgcactt ctgccctttg gaaggtgagt catggcagga tttcctaaga
     1081 aacaacgcaa agagctttag gtgtgcattg ttgagtcata gggacggcgc taaggtccat
     1141 ctaggtacta gaccaaccga aaaacaatac gaaacactgg aaaaccaact agcgttctta
     1201 tgtcagcaag gtttttcttt ggaaaatgcc ttatatgcac tgtccgctgt aggccatttt
     1261 actctgggtt gtgtcttaga agaccaagaa catcaagtag ccaaggagga gagagaaact
     1321 cctaccaccg attctatgcc tcccctttta cgtcaagcga tcgagctgtt tgatcatcaa
     1381 ggggctgaac ccgcctttct ttttggcctt gaattgataa tttgcggtct agagaaacag
     1441 ttaaaatgcg aatccggctc ttagatcgcg tgtattcatc cgctctaacc gaaaaggaag
     1501 gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa
     1561 gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta
     1621 acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgagatcg cgtcagctga
     1681 agcttcgtac gctgcaggtc gacaaccctt aatataactt cgtataatgt atgctatacg
     1741 aagttattag gtctagagat ccgcaggcta accggaacct gtattattta gtttatgcta
     1801 cgttaaataa agacctttcg ttcacataac tgaatgtgta atggccttga gatttcaagc
     1861 ataccaagtt ggtggagacg gggtcgttac aaaagactct ttaagccaag atttccttga
     1921 cagccttggc gatagcatcg ccaacctcag tggtagagtt agaaccacca aggtcaccgg
     1981 ttctgacacc tgcatccaag acatttctaa cagcttcttc aagagcccta ccttcttcaa
     2041 ccaaatccaa ggataacttc aacatcatag ctgcagataa gatggtagca attgggttaa
     2101 ccttgtttgc tggtaaatct ggggcagaac catgacatgg ttcgtacaaa ccgaatgcct
     2161 tgttagtgtc aggtagggaa gctagagatg cagaaggtaa taaacccaaa gaacctggaa
     2221 taacagaggc ttcatcggag ataatatcac caaacatgtt gttggtaata acaacaccgt
     2281 ttagcttagt tggtgattta accaaaatca tagcagcaga gtcgatcaat tggtgctgaa
     2341 cagttaattg tgggaactca gtcttgatgg tttcttcaac agtctttctc cacaatctgg
     2401 aagaggcaag cacgttagcc ttgtcaagag accagattgg taatggtggg ttttgttgca
     2461 atgccaagaa agcagccatt cttgtaattc tttgaacttc aggaacactg tatttctcag
     2521 agtcccaagc aactccgtca ccttcatctt cttttctttc accaaagtag ataccaccaa
     2581 ccaattctct aacgacgacg aaatcggtac cctttgcata ttcaggcttc aaaggagaaa
     2641 gatctagtaa agaatcagaa gcaaagttac atggtctcaa gttggcgtat agacccaatt
     2701 cctttctgat cttcaataga ccttgttctg gtctaactgc gcccgtaccc cattttggac
     2761 caccaacagc acctagtaag acagcatcgg ctttcttaga ggcttctaga gcttcatctg
     2821 gtaaaggagt gccagtggca tcgatggcag cacccccgat caagtgatgt tggaaattga
     2881 acttaatttc tggacggact tcagcaatgg cattcaagac cttaatagct tcgtcagtaa
     2941 cttctttacc gacgtgatca cccggtagga caacgatatt cttagacatg gtataatctg
     3001 tgtagtgtgg gatacttttt acttcttcaa ataggtatca acaataaaaa taaatcgaag
     3061 caaatgtagg aatgcgttaa agcagatgta cttacttaga gtacatagat atatttatat
     3121 aattcaatat ataaaagtat atgaagcatc tgatgttgaa cctgtcatga ctctaaactg
     3181 gaatgggtag ttatgcttct gaaggttttc cgttagaaaa tgagtgcagg gatcaggccc
     3241 ggaaccggtt ttagcctgaa aggaaaagaa aaaaatgcgg tgattactag cacgtgactg
     3301 cgctgaattg gaatctacca caagtacagc gaatggcaag agggaagggg aatatattgt
     3361 gcctggcggt agcgtctgct atatcaaatg ttaaacataa ctaaatgcag tagtgaatag
     3421 agtaatgtat ttctacatat gtgggtaggg ccaatgggag cccgatgtct attgtacaga
     3481 tattttcctt ttatagttaa tcaattcaac ttggaaatca ttgagtcgat atctacttgg
     3541 ataactggag ccgtggcagc gtttgaaatt tgacgagtct tcttgttttg ttcgcgcatt
     3601 ttcagtttat tcaaacggga gatggcaccg aatccactga cttctctgtt caatttgaat
     3661 ggtaatagga agaaagtaca tttcgagaaa ttttcgccca acgtcgaaat ctcatggatg
     3721 atatcctcga tacaaatgac accttcatca cctaatttct cttcaacgat attgttgtca
     3781 ttcaagatta tctcttttgg ttctttatct tctggccttt gccacattat tctagatctc
     3841 ttttggatta aagaacggat agatgcaagt gaaggttgtc cgataacaat atatggtgat
     3901 acaagtttca acaatgggta cacgtccttg gtaagtttga tgaaaacacc agtgttcaac
     3961 tcaacaagtc tcaaaagaga taacactttg taagcctttg ctgggatctt cacagcgagc
     4021 tctcgagaac ccttaatata acttcgtata atgtatgcta tacgaagtta ttaggtgata
     4081 tcagatccac tagtggccta tgcacccaat tcgccctata gtgagtcgta ttacgcgcgc
     4141 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccctgcagga
     4201 ctagtgctga ggcattaatt gtgtccgcgt ttctaaggcc acttttcaat gaaacggata
     4261 ttgatatgct agtaaaagga cgagctcaag agcgaaaata taagtaaaga attcgagtgc
     4321 acttgtctcc atgcagcaag atttcatatg agtctttttt atctttttac tttttacatt
     4381 acacgatatg cactttatga aaatttaacg aggttggaag ccggataatc aaccaaaatc
     4441 aggcacgaag gcacactcgt atatgcatgt tgttgaaact ctgttacgct gaactaacaa
     4501 tcacacatgt agaggtcacc gggaaaagtt gcgaccccat ggaaggtcga tctcttcgtt
     4561 tggctttgct tggctggcgg cattgcgctt cttcgcttat acccgtctct tgacgctcga
     4621 gctcgttcat tgagatacct ttattcttgc acattttctg gcttttttcg ctactcgggt
     4681 acatgtaatc atgcacacag aaggtgctgt agggtgaaag ttcctttgtg ctgtcgtttg
     4741 tttttaatgc caaactttcc ggtgatcaat aaccacctca ttaagacctc agcgcggccg
     4801 caaatttaaa taaaatgaag tgaagttcct atactttcta gagaatagga acttctatag
     4861 tgagtcgaat aagggcgaca caaaatttat tctaaatgca taataaatac tgataacatc
     4921 ttatagtttg tattatattt tgtattatcg ttgacatgta taattttgat atcaaaaact
     4981 gattttccct ttattatttt cgagatttat tttcttaatt ctctttaaca aactagaaat
     5041 attgtatata caaaaaatca taaataatag atgaatagtt taattatagg tgttcatcaa
     5101 tcgaaaaagc aacgtatctt atttaaagtg cgttgctttt ttctcattta taaggttaaa
     5161 taattctcat atatcaagca aagtgacagg cgcccttaaa tattctgaca aatgctcttt
     5221 ccctaaactc cccccataaa aaaacccgcc gaagcgggtt tttacgttat ttgcggatta
     5281 acgattactc gttatcagaa ccgcccaggg ggcccgagct taagactggc cgtcgtttta
     5341 caacacagaa agagtttgta gaaacgcaaa aaggccatcc gtcaggggcc ttctgcttag
     5401 tttgatgcct ggcagttccc tactctcgcc ttccgcttcc tcgctcactg actcgctgcg
     5461 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5521 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5581 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5641 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5701 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5761 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5821 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5881 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5941 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6001 cggtgctaca gagttcttga agtggtgggc taactacggc tacactagaa gaacagtatt
     6061 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6121 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6181 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6241 gaacgacgcg cgcgtaactc acgttaaggg attttggtca tgagcttgcg ccgtcccgtc
     6301 aagtcagcgt aatgctctgc ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6361 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6421 gagggcttac catctggccc cagcgctgcg atgataccgc gagaaccacg ctcaccggct
     6481 ccggatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6541 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6601 ccagttaata gtttgcgcaa cgttgttgcc atcgctacag gcatcgtggt gtcacgctcg
     6661 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6721 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6781 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6841 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6901 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6961 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7021 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7081 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7141 aaaaagggaa taagggcgac acggaaatgt tgaatactca tattcttcct ttttcaatat
     7201 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7261 aaaaataaac aaataggggt cagtgttaca accaattaac caattctgaa cattatcgcg
     7321 agcccattta tacctgaata tggctcataa caccccttgt ttgcctggcg gcagtagcgc
     7381 ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag
     7441 tgtggggact ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc
     7501 agtcgaaaga ctgggccttt cgcccgggct aattatgggg tgtcgccctt attcgactct
     7561 atagtgaagt tcctattctc tagaaagtat aggaacttct gaagtgggga tttaaatgcg
     7621 gccgcgctga gggtttaatc gagatctttg tgttcggtta cccggctcag atcctaactt
     7681 cttcttttgg tatgtttatt cgtataagtt actgttgtcc acaggcaata ctctgcagaa
     7741 aattaaaacg gcattaatgc taggacaacc agaattgtta ctactgtatg tgcgatagtt
     7801 gataactgca acattatgcc cggtatattc tcaaaaaacc ctattactgc atacgaagaa
     7861 atcgcaagag aaatctttcg gtttggaaaa gctcactgtg aggttccttg gagccaatag
     7921 taatacagca caatccaagg aaaaatctgg cctatatgca aggaaggaga gatagtcaaa
     7981 agcattcttt cccctagaag ttggtgcata tatggcatcg ttaaaacata ttacccccaa
     8041 aatttcttct ctaaacgatg
//
