LOCUS       sequence-320164-        6945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  (parent plasmid pCFB262) pXII - 4 -LoxP -SpHIS5 -209bp - 
            pCYC1_BenO_183 -yeGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6945)
  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 6945)
  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: pFA271
FEATURES             Location/Qualifiers
     source          1..6945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(261..426)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     347..371
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             791..1507
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1521..1710
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     protein_bind    1758..1791
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            1846..3046
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        1846..2189
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2190..2843
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his5"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=S. pombe his5
                     /note="yeast auxotrophic marker; corresponds to S. 
                     cerevisiae HIS3"
                     /translation="MGRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHANQ
                     KGEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQ
                     AMGNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLY
                     SFSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     terminator      2849..3046
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    3106..3139
                     /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(3182..3201)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3183..3201)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3211..3227)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3220..3242)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    complement(3847..3880)
                     /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(4350..4366)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4465..4482)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4636..5224)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4716..4735)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5346..6137,6138..6206))
                     /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     5969..5988
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6207..6298)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6340..6359
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6365..6384
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    6591..6624
                     /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 cgcatggttt cagaaagcag agcggaacga ttccaaatta gtggccttgt gctttgcatg
       61 tcaattgtgt taccttcagc tcgtggattt gttttatcaa tacacagtct acagtcaaga
      121 atttttttta tcaaattttg cgttcgagcg tataaaatag ccgctgtagc tacttaagtt
      181 cctgttcagc gatagttttt ttccatcaca cgtactatgg caattaagtc ctcagcgagc
      241 tcgcatggaa tgcgtgcgat gagcgacctc atgctatacc tgagaaagca acctgaccta
      301 caggaaagag ttactcaaga ataagaattt tcgttttaaa acctaagagt cactttaaaa
      361 tttgtataca cttatttttt ttataactta tttaataata aaaatcataa atcataagaa
      421 attcgcttat ttagaagtgt caacaacgta tctaccaacg gaatacgtgc gataccaacg
      481 gaatgcgtgc gatccaggca actttagtgc tgacacataa tactccatag gtattttatt
      541 atacaaataa tgtgtttgaa cttattaaaa cattctttta aggtataaac aacaggcata
      601 tatatatgtg tgcgacgaca catgatcata tggcatgcat gtgctctgta tgtatataaa
      661 actcttgttt tcttcttttc tctaaatatt ctttccttat acattaggac ctttgcagca
      721 taaattacta tacttctata gacacacaaa cacaaataca cacactaaat taataatctg
      781 tcataaaaca atgtctaaag gtgaagaatt attcactggt gttgtcccaa ttttggttga
      841 attagatggt gatgttaatg gtcacaaatt ttctgtctcc ggtgaaggtg aaggtgatgc
      901 tacttacggt aaattgacct taaaatttat ttgtactact ggtaaattgc cagttccatg
      961 gccaacctta gtcactactt tcggttatgg tgttcaatgt tttgcgagat acccagatca
     1021 tatgaaacaa catgactttt tcaagtctgc catgccagaa ggttatgttc aagaaagaac
     1081 tatttttttc aaagatgacg gtaactacaa gaccagagct gaagtcaagt ttgaaggtga
     1141 taccttagtt aatagaatcg aattaaaagg tattgatttt aaagaagatg gtaacatttt
     1201 aggtcacaaa ttggaataca actataactc tcacaatgtt tacatcatgg ctgacaaaca
     1261 aaagaatggt atcaaagtta acttcaaaat tagacacaac attgaagatg gttctgttca
     1321 attagctgac cattatcaac aaaatactcc aattggtgat ggtccagtct tgttaccaga
     1381 caaccattac ttatccactc aatctgcctt atccaaagat ccaaacgaaa agagagacca
     1441 catggtcttg ttagaatttg ttactgctgc tggtattacc catggtatgg atgaattgta
     1501 caaataaatc gcgtgcattc atccgctcta accgaaaagg aaggagttag acaacctgaa
     1561 gtctaggtcc ctatttattt ttttatagtt atgttagtat taagaacgtt atttatattt
     1621 caaatttttc ttttttttct gtacagacgc gtgtacgcat gtaacattat actgaaaacc
     1681 ttgcttgaga aggttttggg acgctcgaag atcgcgtcag ctgaagcttc gtacgctgca
     1741 ggtcgacaac ccttaatata acttcgtata atgtatgcta tacgaagtta ttaggtctag
     1801 agatctgttt agcttgcctc gtccccgccg ggtcacccgg ccagcgacat ggaggcccag
     1861 aataccctcc ttgacagtct tgacgtgcgc agctcagggg catgatgtga ctgtcgcccg
     1921 tacatttagc ccatacatcc ccatgtataa tcatttgcat ccatacattt tgatggccgc
     1981 acggcgcgaa gcaaaaatta cggctcctcg ctgcagacct gcgagcaggg aaacgctccc
     2041 ctcacagacg cgttgaattg tccccacgcc gcgcccctgt agagaaatat aaaaggttag
     2101 gatttgccac tgaggttctt ctttcatata cttcctttta aaatcttgct aggatacagt
     2161 tctcacatca catccgaaca taaacaacca tgggtaggag ggcttttgta gaaagaaata
     2221 cgaacgaaac gaaaatcagc gttgccatcg ctttggacaa agctccctta cctgaagagt
     2281 cgaattttat tgatgaactt ataacttcca agcatgcaaa ccaaaaggga gaacaagtaa
     2341 tccaagtaga cacgggaatt ggattcctgg atcacatgta tcatgcactg gctaaacatg
     2401 caggctggag cttacgactt tactcaagag gtgatttaat catcgatgat catcacactg
     2461 cagaagatac tgctattgca cttggtattg cattcaagca ggctatgggt aactttgccg
     2521 gcgttaaaag atttggacat gcttattgtc cacttgacga agccctttct agaagcgtag
     2581 ttgacttgtc gggacggccc tatgctgtta tcgatttggg attaaagcgt gaaaaggttg
     2641 gggaattgtc ctgtgaaatg atccctcact tactatattc cttttcggta gcagctggaa
     2701 ttactttgca tgttacctgc ttatatggta gtaatgacca tcatcgtgct gaaagcgctt
     2761 ttaaatctct ggctgttgcc atgcgcgcgg ctactagtct tactggaagt tctgaagtcc
     2821 caagcacgaa gggagtgttg taaagagtac tgacaataaa aagattcttg ttttcaagaa
     2881 cttgtcattt gtatagtttt tttatattgt agttgttcta ttttaatcaa atgttagcgt
     2941 gatttatatt ttttttcgcc tcgacatcat ctgcccagat gcgaagttaa gtgcgcagaa
     3001 agtaatatca tgcgtcaatc gtatgtgaat gctggtcgct atactgctgt cgattcgata
     3061 ctaacgccgc catccagttt aaacgagctc tcgagaaccc ttaatataac ttcgtataat
     3121 gtatgctata cgaagttatt aggtgatatc agatccacta gtggcctatg cacccaattc
     3181 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
     3241 ggaaaaccct ggcgttaccc ctgcaggact agtgctgagg cattaatcca actttgtact
     3301 attccttccc cgtttactca attcttgaag ccaatttgta caattcccca ttagagtcaa
     3361 ataaaaggat gcctcacgga ggtatgttac ccgcgctatt tcacatggct cattgaatta
     3421 gaggtggaat ttggtgtacc ctcccctcct catctgatga agtagtgatc cgacaattct
     3481 taaaagttgt agacattact tttaccacca actaagttgt atttatattg ctacccttat
     3541 ccttttatat ctaactagcg ctcataaggt tggggcaata ctaaaactgt gttcttattc
     3601 aactcattaa atacgtggca gtacgtaccc tattagaaac aataggaaac agcagagtcg
     3661 gaagaagcca aatgccagat ttgaagtcca aaaccttgtc aagccaatct ttgggagcgg
     3721 ctattcctcc agaaattgtg taccaaatac ttacatacca gtttagggat ttgttaagaa
     3781 atgaccatcc aggtacggca gaaaattaag acctcagcgc ggccgcaaat ttaaataaaa
     3841 tgaagtgaag ttcctatact ttctagagaa taggaacttc tatagtgagt cgaataaggg
     3901 cgacacaaaa tttattctaa atgcataata aatactgata acatcttata gtttgtatta
     3961 tattttgtat tatcgttgac atgtataatt ttgatatcaa aaactgattt tccctttatt
     4021 attttcgaga tttattttct taattctctt taacaaacta gaaatattgt atatacaaaa
     4081 aatcataaat aatagatgaa tagtttaatt ataggtgttc atcaatcgaa aaagcaacgt
     4141 atcttattta aagtgcgttg cttttttctc atttataagg ttaaataatt ctcatatatc
     4201 aagcaaagtg acaggcgccc ttaaatattc tgacaaatgc tctttcccta aactcccccc
     4261 ataaaaaaac ccgccgaagc gggtttttac gttatttgcg gattaacgat tactcgttat
     4321 cagaaccgcc cagggggccc gagcttaaga ctggccgtcg ttttacaaca cagaaagagt
     4381 ttgtagaaac gcaaaaaggc catccgtcag gggccttctg cttagtttga tgcctggcag
     4441 ttccctactc tcgccttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4501 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4561 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4621 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4681 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4741 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4801 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4861 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4921 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4981 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5041 cttgaagtgg tgggctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     5101 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5161 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5221 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg acgcgcgcgt
     5281 aactcacgtt aagggatttt ggtcatgagc ttgcgccgtc ccgtcaagtc agcgtaatgc
     5341 tctgcttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5401 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5461 ggccccagcg ctgcgatgat accgcgagaa ccacgctcac cggctccgga tttatcagca
     5521 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5581 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5641 cgcaacgttg ttgccatcgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5701 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5761 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5821 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5881 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5941 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6001 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6061 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6121 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6181 gcgacacgga aatgttgaat actcatattc ttcctttttc aatattattg aagcatttat
     6241 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6301 ggggtcagtg ttacaaccaa ttaaccaatt ctgaacatta tcgcgagccc atttatacct
     6361 gaatatggct cataacaccc cttgtttgcc tggcggcagt agcgcggtgg tcccacctga
     6421 ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggactcccca
     6481 tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     6541 cctttcgccc gggctaatta tggggtgtcg cccttattcg actctatagt gaagttccta
     6601 ttctctagaa agtataggaa cttctgaagt ggggatttaa atgcggccgc gctgagggtt
     6661 taatgtatcc ggctgttcct tcatagccct ttcaatgaac gttgcagccc tttgaagatt
     6721 ggccattttg tcaggactcg agcctgacag ttggaccaac gcaactttaa ttttttgtga
     6781 aagaatcttc gaagcactca tactggcgat cttcacgccc tcctgctatt acaaaagctg
     6841 tgtttttaca agaatcaaat taagttagca agatattata caacattatt gataatttca
     6901 atatcgtgtt cgtacctgat gacgtatctg tgcattgata aggcc
//
