LOCUS       Exported                6925 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  (parent plasmid pCFB262) pXII - 4 -LoxP -SpHIS5 -209bp - 
            pCYC1_BenO_182 -yeGFP.
ACCESSION   .
VERSION     .
KEYWORDS    pFA272
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6925)
  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 6925)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6925
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      420..585
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     506..530
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             930..1646
                     /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      1660..1849
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     protein_bind    complement(1897..1930)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     gene            1985..3185
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        1985..2328
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2329..2982
                     /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      2988..3185
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    complement(3245..3278)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     primer_bind     complement(3321..3340)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3322..3340)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3350..3366)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3359..3381)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     protein_bind    complement(3986..4019)
                     /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(4489..4505)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4604..4621)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4775..5363)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4855..4874)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5485..6345)
                     /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     6108..6127
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6346..6437)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6479..6498
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6504..6523
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    6730..6763
                     /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 cttcgaagca ctcatactgg cgatcttcac gccctcctgc tattacaaaa gctgtgtttt
       61 tacaagaatc aaattaagtt agcaagatat tatacaacat tattgataat ttcaatatcg
      121 tgttcgtacc tgatgacgta tctgtgcatt gataaggccc gcatggtttc agaaagcaga
      181 gcggaacgat tccaaattag tggccttgtg ctttgcatgt caattgtgtt accttcagct
      241 cgtggatttg ttttatcaat acacagtcta cagtcaagaa ttttttttat caaattttgc
      301 gttcgagcgt ataaaatagc cgctgtagct acttaagttc ctgttcagcg atagtttttt
      361 tccatcacac gtactatggc aattaagtcc tcagcgagct cgcatggaat gcgtgcgatg
      421 agcgacctca tgctatacct gagaaagcaa cctgacctac aggaaagagt tactcaagaa
      481 taagaatttt cgttttaaaa cctaagagtc actttaaaat ttgtatacac ttattttttt
      541 tataacttat ttaataataa aaatcataaa tcataagaaa ttcgcttatt tagaagtgtc
      601 aacaacgtat ctaccaacgg aatgcgtgcg atccaggcaa ctttagtgct gacacataca
      661 tactccatag gtattttatt atacaaataa tgtgtttgaa cttattaaaa cattctttta
      721 aggtataaac aaaggcatat atatatgtgt gcgacgacac atgatcatat ggcatgcatg
      781 tgctctgtat gtatataaaa ctcttgtttt cttcttttct ctaaatattc tttccttata
      841 cattaggacc tttgcagcat aaattactat acttctatag acacacaaac acaaatacac
      901 acactaaatt aataatctgt cataaaacaa tgtctaaagg tgaagaatta ttcactggtg
      961 ttgtcccaat tttggttgaa ttagatggtg atgttaatgg tcacaaattt tctgtctccg
     1021 gtgaaggtga aggtgatgct acttacggta aattgacctt aaaatttatt tgtactactg
     1081 gtaaattgcc agttccatgg ccaaccttag tcactacttt cggttatggt gttcaatgtt
     1141 ttgcgagata cccagatcat atgaaacaac atgacttttt caagtctgcc atgccagaag
     1201 gttatgttca agaaagaact atttttttca aagatgacgg taactacaag accagagctg
     1261 aagtcaagtt tgaaggtgat accttagtta atagaatcga attaaaaggt attgatttta
     1321 aagaagatgg taacatttta ggtcacaaat tggaatacaa ctataactct cacaatgttt
     1381 acatcatggc tgacaaacaa aagaatggta tcaaagttaa cttcaaaatt agacacaaca
     1441 ttgaagatgg ttctgttcaa ttagctgacc attatcaaca aaatactcca attggtgatg
     1501 gtccagtctt gttaccagac aaccattact tatccactca atctgcctta tccaaagatc
     1561 caaacgaaaa gagagaccac atggtcttgt tagaatttgt tactgctgct ggtattaccc
     1621 atggtatgga tgaattgtac aaataaatcg cgtgcattca tccgctctaa ccgaaaagga
     1681 aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta tgttagtatt
     1741 aagaacgtta tttatatttc aaatttttct tttttttctg tacagacgcg tgtacgcatg
     1801 taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaaga tcgcgtcagc
     1861 tgaagcttcg tacgctgcag gtcgacaacc cttaatataa cttcgtataa tgtatgctat
     1921 acgaagttat taggtctaga gatctgttta gcttgcctcg tccccgccgg gtcacccggc
     1981 cagcgacatg gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc
     2041 atgatgtgac tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc
     2101 catacatttt gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg
     2161 cgagcaggga aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta
     2221 gagaaatata aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa
     2281 aatcttgcta ggatacagtt ctcacatcac atccgaacat aaacaaccat gggtaggagg
     2341 gcttttgtag aaagaaatac gaacgaaacg aaaatcagcg ttgccatcgc tttggacaaa
     2401 gctcccttac ctgaagagtc gaattttatt gatgaactta taacttccaa gcatgcaaac
     2461 caaaagggag aacaagtaat ccaagtagac acgggaattg gattcctgga tcacatgtat
     2521 catgcactgg ctaaacatgc aggctggagc ttacgacttt actcaagagg tgatttaatc
     2581 atcgatgatc atcacactgc agaagatact gctattgcac ttggtattgc attcaagcag
     2641 gctatgggta actttgccgg cgttaaaaga tttggacatg cttattgtcc acttgacgaa
     2701 gccctttcta gaagcgtagt tgacttgtcg ggacggccct atgctgttat cgatttggga
     2761 ttaaagcgtg aaaaggttgg ggaattgtcc tgtgaaatga tccctcactt actatattcc
     2821 ttttcggtag cagctggaat tactttgcat gttacctgct tatatggtag taatgaccat
     2881 catcgtgctg aaagcgcttt taaatctctg gctgttgcca tgcgcgcggc tactagtctt
     2941 actggaagtt ctgaagtccc aagcacgaag ggagtgttgt aaagagtact gacaataaaa
     3001 agattcttgt tttcaagaac ttgtcatttg tatagttttt ttatattgta gttgttctat
     3061 tttaatcaaa tgttagcgtg atttatattt tttttcgcct cgacatcatc tgcccagatg
     3121 cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg tatgtgaatg ctggtcgcta
     3181 tactgctgtc gattcgatac taacgccgcc atccagttta aacgagctct cgagaaccct
     3241 taatataact tcgtataatg tatgctatac gaagttatta ggtgatatca gatccactag
     3301 tggcctatgc acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg
     3361 ttttacaacg tcgtgactgg gaaaaccctg gcgttacccc tgcaggacta gtgctgaggc
     3421 attaatccaa ctttgtacta ttccttcccc gtttactcaa ttcttgaagc caatttgtac
     3481 aattccccat tagagtcaaa taaaaggatg cctcacggag gtatgttacc cgcgctattt
     3541 cacatggctc attgaattag aggtggaatt tggtgtaccc tcccctcctc atctgatgaa
     3601 gtagtgatcc gacaattctt aaaagttgta gacattactt ttaccaccaa ctaagttgta
     3661 tttatattgc tacccttatc cttttatatc taactagcgc tcataaggtt ggggcaatac
     3721 taaaactgtg ttcttattca actcattaaa tacgtggcag tacgtaccct attagaaaca
     3781 ataggaaaca gcagagtcgg aagaagccaa atgccagatt tgaagtccaa aaccttgtca
     3841 agccaatctt tgggagcggc tattcctcca gaaattgtgt accaaatact tacataccag
     3901 tttagggatt tgttaagaaa tgaccatcca ggtacggcag aaaattaaga cctcagcgcg
     3961 gccgcaaatt taaataaaat gaagtgaagt tcctatactt tctagagaat aggaacttct
     4021 atagtgagtc gaataagggc gacacaaaat ttattctaaa tgcataataa atactgataa
     4081 catcttatag tttgtattat attttgtatt atcgttgaca tgtataattt tgatatcaaa
     4141 aactgatttt ccctttatta ttttcgagat ttattttctt aattctcttt aacaaactag
     4201 aaatattgta tatacaaaaa atcataaata atagatgaat agtttaatta taggtgttca
     4261 tcaatcgaaa aagcaacgta tcttatttaa agtgcgttgc ttttttctca tttataaggt
     4321 taaataattc tcatatatca agcaaagtga caggcgccct taaatattct gacaaatgct
     4381 ctttccctaa actcccccca taaaaaaacc cgccgaagcg ggtttttacg ttatttgcgg
     4441 attaacgatt actcgttatc agaaccgccc agggggcccg agcttaagac tggccgtcgt
     4501 tttacaacac agaaagagtt tgtagaaacg caaaaaggcc atccgtcagg ggccttctgc
     4561 ttagtttgat gcctggcagt tccctactct cgccttccgc ttcctcgctc actgactcgc
     4621 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4681 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4741 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4801 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4861 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4921 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4981 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5041 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5101 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5161 taggcggtgc tacagagttc ttgaagtggt gggctaacta cggctacact agaagaacag
     5221 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5281 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5341 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5401 agtggaacga cgcgcgcgta actcacgtta agggattttg gtcatgagct tgcgccgtcc
     5461 cgtcaagtca gcgtaatgct ctgcttacca atgcttaatc agtgaggcac ctatctcagc
     5521 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5581 acgggagggc ttaccatctg gccccagcgc tgcgatgata ccgcgagaac cacgctcacc
     5641 ggctccggat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5701 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5761 ttcgccagtt aatagtttgc gcaacgttgt tgccatcgct acaggcatcg tggtgtcacg
     5821 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5881 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5941 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6001 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6061 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6121 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6181 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6241 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6301 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatattct tcctttttca
     6361 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6421 ttagaaaaat aaacaaatag gggtcagtgt tacaaccaat taaccaattc tgaacattat
     6481 cgcgagccca tttatacctg aatatggctc ataacacccc ttgtttgcct ggcggcagta
     6541 gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg
     6601 gtagtgtggg gactccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag
     6661 gctcagtcga aagactgggc ctttcgcccg ggctaattat ggggtgtcgc ccttattcga
     6721 ctctatagtg aagttcctat tctctagaaa gtataggaac ttctgaagtg gggatttaaa
     6781 tgcggccgcg ctgagggttt aatgtatccg gctgttcctt catagccctt tcaatgaacg
     6841 ttgcagccct ttgaagattg gccattttgt caggactcga gcctgacagt tggaccaacg
     6901 caactttaat tttttgtgaa agaat
//
