LOCUS       sequence-320162-        6932 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  (parent plasmid pCFB262) pXII - 4 -LoxP -SpHIS5 -209bp - pCYC1 -PcaO
            -108 -yeGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6932)
  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 6932)
  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: pFA197
FEATURES             Location/Qualifiers
     source          1..6932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(178..343)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     264..288
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             695..1411
                     /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      1425..1614
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     protein_bind    1662..1695
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            1750..2950
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        1750..2093
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2094..2747
                     /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      2753..2950
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    3010..3043
                     /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(3086..3105)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3087..3105)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3115..3131)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3124..3146)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    complement(3751..3784)
                     /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(4254..4270)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4369..4386)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4540..5128)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4620..4639)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5250..6041,6042..6110))
                     /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     5873..5892
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6111..6202)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6244..6263
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6269..6288
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    6495..6528
                     /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 tggatttgtt ttatcaatac acagtctaca gtcaagaatt ttttttatca aattttgcgt
       61 tcgagcgtat aaaatagccg ctgtagctac ttaagttcct gttcagcgat agtttttttc
      121 catcacacgt actatggcaa ttaagtcctc agcgagctcg catggaatgc gtgcgatgag
      181 cgacctcatg ctatacctga gaaagcaacc tgacctacag gaaagagtta ctcaagaata
      241 agaattttcg ttttaaaacc taagagtcac tttaaaattt gtatacactt atttttttta
      301 taacttattt aataataaaa atcataaatc ataagaaatt cgcttattta gaagtgtcaa
      361 caacgtatct accaacggaa tacgtgcgat ccaggcaact ttagtgctga cacatacagg
      421 catatatata tgtgtgcgac gacacatgat catatggcat gcatgtgctc tgtatgtata
      481 taaaactctt ggatcgtata acctcctggt taagggaaag ccacgaaata tcattttacc
      541 taaccggatg aaacatccaa atctgacgac gttttcttct tttctctaaa tattctttcc
      601 ttatacatta ggacctttgc agcataaatt actatacttc tatagacaca caaacacaaa
      661 tacacacact aaattaataa tctgtcataa aacaatgtct aaaggtgaag aattattcac
      721 tggtgttgtc ccaattttgg ttgaattaga tggtgatgtt aatggtcaca aattttctgt
      781 ctccggtgaa ggtgaaggtg atgctactta cggtaaattg accttaaaat ttatttgtac
      841 tactggtaaa ttgccagttc catggccaac cttagtcact actttcggtt atggtgttca
      901 atgttttgcg agatacccag atcatatgaa acaacatgac tttttcaagt ctgccatgcc
      961 agaaggttat gttcaagaaa gaactatttt tttcaaagat gacggtaact acaagaccag
     1021 agctgaagtc aagtttgaag gtgatacctt agttaataga atcgaattaa aaggtattga
     1081 ttttaaagaa gatggtaaca ttttaggtca caaattggaa tacaactata actctcacaa
     1141 tgtttacatc atggctgaca aacaaaagaa tggtatcaaa gttaacttca aaattagaca
     1201 caacattgaa gatggttctg ttcaattagc tgaccattat caacaaaata ctccaattgg
     1261 tgatggtcca gtcttgttac cagacaacca ttacttatcc actcaatctg ccttatccaa
     1321 agatccaaac gaaaagagag accacatggt cttgttagaa tttgttactg ctgctggtat
     1381 tacccatggt atggatgaat tgtacaaata aatcgcgtgc attcatccgc tctaaccgaa
     1441 aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta
     1501 gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacag acgcgtgtac
     1561 gcatgtaaca ttatactgaa aaccttgctt gagaaggttt tgggacgctc gaagatcgcg
     1621 tcagctgaag cttcgtacgc tgcaggtcga caacccttaa tataacttcg tataatgtat
     1681 gctatacgaa gttattaggt ctagagatct gtttagcttg cctcgtcccc gccgggtcac
     1741 ccggccagcg acatggaggc ccagaatacc ctccttgaca gtcttgacgt gcgcagctca
     1801 ggggcatgat gtgactgtcg cccgtacatt tagcccatac atccccatgt ataatcattt
     1861 gcatccatac attttgatgg ccgcacggcg cgaagcaaaa attacggctc ctcgctgcag
     1921 acctgcgagc agggaaacgc tcccctcaca gacgcgttga attgtcccca cgccgcgccc
     1981 ctgtagagaa atataaaagg ttaggatttg ccactgaggt tcttctttca tatacttcct
     2041 tttaaaatct tgctaggata cagttctcac atcacatccg aacataaaca accatgggta
     2101 ggagggcttt tgtagaaaga aatacgaacg aaacgaaaat cagcgttgcc atcgctttgg
     2161 acaaagctcc cttacctgaa gagtcgaatt ttattgatga acttataact tccaagcatg
     2221 caaaccaaaa gggagaacaa gtaatccaag tagacacggg aattggattc ctggatcaca
     2281 tgtatcatgc actggctaaa catgcaggct ggagcttacg actttactca agaggtgatt
     2341 taatcatcga tgatcatcac actgcagaag atactgctat tgcacttggt attgcattca
     2401 agcaggctat gggtaacttt gccggcgtta aaagatttgg acatgcttat tgtccacttg
     2461 acgaagccct ttctagaagc gtagttgact tgtcgggacg gccctatgct gttatcgatt
     2521 tgggattaaa gcgtgaaaag gttggggaat tgtcctgtga aatgatccct cacttactat
     2581 attccttttc ggtagcagct ggaattactt tgcatgttac ctgcttatat ggtagtaatg
     2641 accatcatcg tgctgaaagc gcttttaaat ctctggctgt tgccatgcgc gcggctacta
     2701 gtcttactgg aagttctgaa gtcccaagca cgaagggagt gttgtaaaga gtactgacaa
     2761 taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata ttgtagttgt
     2821 tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca tcatctgccc
     2881 agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt gaatgctggt
     2941 cgctatactg ctgtcgattc gatactaacg ccgccatcca gtttaaacga gctctcgaga
     3001 acccttaata taacttcgta taatgtatgc tatacgaagt tattaggtga tatcagatcc
     3061 actagtggcc tatgcaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc
     3121 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccctgcag gactagtgct
     3181 gaggcattaa tccaactttg tactattcct tccccgttta ctcaattctt gaagccaatt
     3241 tgtacaattc cccattagag tcaaataaaa ggatgcctca cggaggtatg ttacccgcgc
     3301 tatttcacat ggctcattga attagaggtg gaatttggtg taccctcccc tcctcatctg
     3361 atgaagtagt gatccgacaa ttcttaaaag ttgtagacat tacttttacc accaactaag
     3421 ttgtatttat attgctaccc ttatcctttt atatctaact agcgctcata aggttggggc
     3481 aatactaaaa ctgtgttctt attcaactca ttaaatacgt ggcagtacgt accctattag
     3541 aaacaatagg aaacagcaga gtcggaagaa gccaaatgcc agatttgaag tccaaaacct
     3601 tgtcaagcca atctttggga gcggctattc ctccagaaat tgtgtaccaa atacttacat
     3661 accagtttag ggatttgtta agaaatgacc atccaggtac ggcagaaaat taagacctca
     3721 gcgcggccgc aaatttaaat aaaatgaagt gaagttccta tactttctag agaataggaa
     3781 cttctatagt gagtcgaata agggcgacac aaaatttatt ctaaatgcat aataaatact
     3841 gataacatct tatagtttgt attatatttt gtattatcgt tgacatgtat aattttgata
     3901 tcaaaaactg attttccctt tattattttc gagatttatt ttcttaattc tctttaacaa
     3961 actagaaata ttgtatatac aaaaaatcat aaataataga tgaatagttt aattataggt
     4021 gttcatcaat cgaaaaagca acgtatctta tttaaagtgc gttgcttttt tctcatttat
     4081 aaggttaaat aattctcata tatcaagcaa agtgacaggc gcccttaaat attctgacaa
     4141 atgctctttc cctaaactcc ccccataaaa aaacccgccg aagcgggttt ttacgttatt
     4201 tgcggattaa cgattactcg ttatcagaac cgcccagggg gcccgagctt aagactggcc
     4261 gtcgttttac aacacagaaa gagtttgtag aaacgcaaaa aggccatccg tcaggggcct
     4321 tctgcttagt ttgatgcctg gcagttccct actctcgcct tccgcttcct cgctcactga
     4381 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4441 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4501 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4561 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4621 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4681 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4741 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4801 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4861 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4921 gtatgtaggc ggtgctacag agttcttgaa gtggtgggct aactacggct acactagaag
     4981 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5041 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5101 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5161 cgctcagtgg aacgacgcgc gcgtaactca cgttaaggga ttttggtcat gagcttgcgc
     5221 cgtcccgtca agtcagcgta atgctctgct taccaatgct taatcagtga ggcacctatc
     5281 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5341 acgatacggg agggcttacc atctggcccc agcgctgcga tgataccgcg agaaccacgc
     5401 tcaccggctc cggatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5461 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5521 agtagttcgc cagttaatag tttgcgcaac gttgttgcca tcgctacagg catcgtggtg
     5581 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5641 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5701 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5761 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5821 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5881 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5941 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6001 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6061 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat attcttcctt
     6121 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6181 tgtatttaga aaaataaaca aataggggtc agtgttacaa ccaattaacc aattctgaac
     6241 attatcgcga gcccatttat acctgaatat ggctcataac accccttgtt tgcctggcgg
     6301 cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc
     6361 cgatggtagt gtggggactc cccatgcgag agtagggaac tgccaggcat caaataaaac
     6421 gaaaggctca gtcgaaagac tgggcctttc gcccgggcta attatggggt gtcgccctta
     6481 ttcgactcta tagtgaagtt cctattctct agaaagtata ggaacttctg aagtggggat
     6541 ttaaatgcgg ccgcgctgag ggtttaatgt atccggctgt tccttcatag ccctttcaat
     6601 gaacgttgca gccctttgaa gattggccat tttgtcagga ctcgagcctg acagttggac
     6661 caacgcaact ttaatttttt gtgaaagaat cttcgaagca ctcatactgg cgatcttcac
     6721 gccctcctgc tattacaaaa gctgtgtttt tacaagaatc aaattaagtt agcaagatat
     6781 tatacaacat tattgataat ttcaatatcg tgttcgtacc tgatgacgta tctgtgcatt
     6841 gataaggccc gcatggtttc agaaagcaga gcggaacgat tccaaattag tggccttgtg
     6901 ctttgcatgt caattgtgtt accttcagct cg
//
