LOCUS       sequence-254574-        8403 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses pTEF2-mKate2-tIDP1 and pTRX2_5xUAS-yEGFP-tCYC1 in yeast 
            from plasdmid pRS413.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8403)
  AUTHORS   Zhang J, Sonnenschein N, Pihl TP, Pedersen KR, Jensen MK, Keasling 
            JD
  TITLE     Engineering an NADPH/NADP(+) Redox Biosensor in Yeast.
  JOURNAL   ACS Synth Biol. 2016 Dec 16;5(12):1546-1556. doi: 
            10.1021/acssynbio.6b00135. Epub 2016 Jul 25.
  PUBMED    27419466
REFERENCE   2  (bases 1 to 8403)
  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.6b00135"; journalName: "ACS Synth Biol"; date: 
            "2016-12-16- 16"; volume: "5"; issue: "12"; pages: "1546-1556"
COMMENT     Sequence Label: pCfB4170
FEATURES             Location/Qualifiers
     source          1..8403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1715..2431)
                     /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      3347..3512
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3402..3426)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3540..3559)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3541..3559)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3569..3586)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3569..3585)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3578..3600)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3726..4181
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3813..3832)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4023..4044
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4441..5103)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(5104..5291)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(5438..5457)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5557..5579
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5617..5635)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6217..6321
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6322..6390,6391..7182)
                     /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     complement(6540..6559)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7353..7941
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8095..8112
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8229..8250
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8265..8270,8271..8288,8289..8295)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8303..8319
                     /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)."
     primer_bind     8308..8330
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8327..8343
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8327..8343
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8362..8382
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8364..8382
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 atatatgggg ccgtatactt acatatagta gatgtcaagc gtaggcgctt cccctgccgg
       61 ctgtgagggc gccataacca aggtatctat agaccgccaa tcagcaaact acctccgtac
      121 attcatgttg cacccacaca tttatacacc cagaccgcga caaattaccc ataaggttgt
      181 ttgtgacggc gtcgtacaag agaacgtggg aactttttag gctcaccaaa aaagaaagaa
      241 aaaatacgag ttgctgacag aagcctcaag aaaaaaaaaa ttcttcttcg actatgctgg
      301 aggcagagat gatcgagccg gtagttaact atatatagct aaattggttc catcaccttc
      361 ttttctggtg tcgctccttc tagtgctatt tctggctttt cctatttttt tttttccatt
      421 tttctttctc tctttctaat atataaattc tcttgcattt tctatttttc tctctatcta
      481 ttctacttgt ttattccctt caaggttttt ttttaaggag tacttgtttt tagaatatac
      541 ggtcaacgaa ctataattaa ctaaacatgg tttctgaact catcaaggaa aacatgcaca
      601 tgaaacttta catggaaggt actgtgaaca atcatcattt taagtgtaca tccgagggtg
      661 aaggcaaacc ttacgaagga actcaaacta tgagaattaa agctgtagaa ggtggaccat
      721 taccttttgc atttgatatc ttggcaacat cattcatgta tgggagcaag acattcataa
      781 accatactca aggtatacca gactttttca aacagagttt tccagagggt tttacatggg
      841 aaagagtaac aacgtacgag gatggaggtg tattgacagc cactcaagac acatcacttc
      901 aagatgggtg tttaatctac aatgtcaaga ttagaggcgt caatttccct tctaatggtc
      961 cagttatgca gaaaaagaca ttaggctggg aagcgtcaac cgaaaccctg taccctgctg
     1021 atggtggcct agaaggcaga gctgacatgg cccttaaact ggttggtgga gggcatctaa
     1081 tctgcaattt gaaaaccact tatcgttcta aaaagccagc caaaaaccta aagatgccag
     1141 gtgtttacta cgtcgaccga agattagaaa ggattaaaga ggctgataaa gagacttatg
     1201 ttgaacaaca cgaagtggca gtggctagat actgtgattt gccatctaag ttgggacaca
     1261 gataaagctt cgaatttacg tagcccaatc taccactttt ttttttcatt ttttaaagtg
     1321 ttatacttag ttatgctcta ggataatgaa ctactttttt tttttttttt ttttactgtt
     1381 atcataaata tatatacctt attgttgttt gcaaccgtcg gttaattcct tatcaaggtt
     1441 ccccaagttc ggatcattac catcaatttc caacattttc atgagttctt cttcttcatt
     1501 accgtgtttt agggggctgt tctcgagctc ttctcaagca aggttttcag tataatgtta
     1561 catgcgtaca cgcgtctgta cagaaaaaaa agaaaaattt gaaatataaa taacgttctt
     1621 aatactaaca taactataaa aaaataaata gggacctaga cttcaggttg tctaactcct
     1681 tccttttcgg ttagagcgga tgaatgcacg cgatttattt gtacaattca tccataccat
     1741 gggtaatacc agcagcagta acaaattcta acaagaccat gtggtctctc ttttcgtttg
     1801 gatctttgga taaggcagat tgagtggata agtaatggtt gtctggtaac aagactggac
     1861 catcaccaat tggagtattt tgttgataat ggtcagctaa ttgaacagaa ccatcttcaa
     1921 tgttgtgtct aattttgaag ttaactttga taccattctt ttgtttgtca gccatgatgt
     1981 aaacattgtg agagttatag ttgtattcca atttgtgacc taaaatgtta ccatcttctt
     2041 taaaatcaat accttttaat tcgattctat taactaaggt atcaccttca aacttgactt
     2101 cagctctggt cttgtagtta ccgtcatctt tgaaaaaaat agttctttct tgaacataac
     2161 cttctggcat ggcagacttg aaaaagtcat gttgtttcat atgatctggg tatctcgcaa
     2221 aacattgaac accataaccg aaagtagtga ctaaggttgg ccatggaact ggcaatttac
     2281 cagtagtaca aataaatttt aaggtcaatt taccgtaagt agcatcacct tcaccttcac
     2341 cggagacaga aaatttgtga ccattaacat caccatctaa ttcaaccaaa attgggacaa
     2401 caccagtgaa taattcttca cctttagaca ttgttttatg acagattatt gatgtgttat
     2461 ttaaagatat cgtagactct cgtgtatgtg tgcgtgtata attcttgcta gacaacaaat
     2521 aaaaactggt cctcgttatt ctcttgtcag cttttcatcc cccgaatggc tttcttatat
     2581 actgatatcc ctctttcggc tggctggaaa ctgaacgcgc ttagtaagaa aagagccacc
     2641 ttgtagggag catcctttac taagagtata aacaattaca catatacttt gatgattact
     2701 taatattcac tggcactcaa gatgacagat ctgatatccc tctttcggct ggctggaaac
     2761 tgaacgcgct tagtaagaaa agagccacct tgtagggagc atcctttact aagagtataa
     2821 acaattacac atatactttg atgattactt aatattcact ggcactcaag ctgatatccc
     2881 tctttcggct ggctggaaac tgaacgcgct tagtaagaaa agagccacct tgtagggagc
     2941 atcctttact aagagtataa acaattacac atatactttg atgattactt aatattcact
     3001 ggcactcaag agtgcaggtc tgatatccct ctttcggctg gctggaaact gaacgcgctt
     3061 agtaagaaaa gagccacctt gtagggagca tcctttacta agagtataaa caattacaca
     3121 tatactttga tgattactta atattcactg gcactcaagc tgatatccct ctttcggctg
     3181 gctggaaact gaacgcgctt agtaagaaaa gagccacctt gtagggagca tcctttacta
     3241 agagtataaa caattacaca tatactttga tgattactta atattcactg gcactcaaga
     3301 tcgcacgcat tccgttggta gatacgttgt tgacacttct aaataagcga atttcttatg
     3361 atttatgatt tttattatta aataagttat aaaaaaaata agtgtataca aattttaaag
     3421 tgactcttag gttttaaaac gaaaattctt attcttgagt aactctttcc tgtaggtcag
     3481 gttgctttct caggtatagc atgaggtcgc tctcgagggg gggcccggta cccaattcgc
     3541 cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg
     3601 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     3661 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     3721 aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3781 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3841 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3901 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3961 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4021 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4081 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4141 atttaacgcg aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc
     4201 cttacgcatc tgtgcggtat ttcacaccgc atagatccgt cgagttcaag agaaaaaaaa
     4261 agaaaaagca aaaagaaaaa aggaaagcgc gcctcgttca gaatgacacg tatagaatga
     4321 tgcattacct tgtcatcttc agtatcatac tgttcgtata catacttact gacattcata
     4381 ggtatacata tatacacatg tatatatatc gtatgctgca gctttaaata atcggtgtca
     4441 ctacataaga acacctttgg tggagggaac atcgttggta ccattgggcg aggtggcttc
     4501 tcttatggca accgcaagag ccttgaacgc actctcacta cggtgatgat cattcttgcc
     4561 tcgcagacaa tcaacgtgga gggtaattct gctagcctct gcaaagcttt caagaaaatg
     4621 cgggatcatc tcgcaagaga gatctcctac tttctccctt tgcaaaccaa gttcgacaac
     4681 tgcgtacggc ctgttcgaaa gatctaccac cgctctggaa agtgcctcat ccaaaggcgc
     4741 aaatcctgat ccaaaccttt ttactccacg cacggcccct agggcctctt taaaagcttg
     4801 accgagagca atcccgcagt cttcagtggt gtgatggtcg tctatgtgta agtcaccaat
     4861 gcactcaacg attagcgacc agccggaatg cttggccaga gcatgtatca tatggtccag
     4921 aaaccctata cctgtgtgga cgttaatcac ttgcgattgt gtggcctgtt ctgctactgc
     4981 ttctgcctct ttttctggga agatcgagtg ctctatcgct aggggaccac cctttaaaga
     5041 gatcgcaatc tgaatcttgg tttcatttgt aatacgcttt actagggctt tctgctctgt
     5101 catctttgcc ttcgtttatc ttgcctgctc attttttagt atattcttcg aagaaatcac
     5161 attactttat ataatgtata attcattatg tgataatgcc aatcgctaag aaaaaaaaag
     5221 agtcatccgc taggtggaaa aaaaaaaatg aaaatcatta ccgaggcata aaaaaatata
     5281 gagtgtacta gaggaggcca agagtaatag aaaaagaaaa ttgcgggaaa ggactgtgtt
     5341 atgacttccc tgactaatgc cgtgttcaaa cgatacctgg cagtgactcc tagcgctcac
     5401 caagctctta aaacgggaat ttatggtgca ctctcagtac aatctgctct gatgccgcat
     5461 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     5521 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     5581 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     5641 aggttaatgt catgataata atggtgccac ctgacggatc gcttgcctgt aacttacacg
     5701 cgcctcgtat cttttaatga tggaataatt tgggaattta ctctgtgttt atttattttt
     5761 atgttttgta tttggatttt agaaagtaaa taaagaaggt agaagagtta cggaatgaag
     5821 aaaaaaaaat aaacaaaggt ttaaaaaatt tcaacaaaaa gcgtacttta catatatatt
     5881 tattagacaa gaaaagcaga ttaaatagat atacattcga ttaacgataa gtaaaatgta
     5941 aaatcacagg attttcgtgt gtggtcttct acacagacaa gatgaaacaa ttcggcatta
     6001 atacctgaga gcaggaagag caagataaaa ggtagtattt gttggcgatc cccctagagt
     6061 cttttacatc ttcggaaaac aaaaactatt ttttmtwtaa cttctttatt tactttctaa
     6121 kattaaattc ccaaattatt ccatcattaa aagatacgag gcgcgtgtaa gttacaggca
     6181 agcgatccgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     6241 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     6301 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6361 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6421 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6481 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6541 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     6601 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6661 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6721 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6781 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6841 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6901 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6961 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     7021 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     7081 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     7141 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     7201 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     7261 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     7321 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7381 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7441 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     7501 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7561 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7621 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7681 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7741 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7801 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7861 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7921 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7981 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     8041 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     8101 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     8161 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     8221 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     8281 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     8341 gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctc
     8401 gag
//
