LOCUS       sequence-245887-        7663 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses pTRX2_5xUAS-ALD6-tADH1 in yeast from plasdmid pRS413.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7663)
  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 7663)
  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: pCfB4929
FEATURES             Location/Qualifiers
     source          1..7663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     36..56
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        38..56
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     terminator      2685..2850
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2740..2764)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(2878..2897)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2879..2897)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2907..2924)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2907..2923)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2916..2938)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3064..3519
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3151..3170)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3361..3382
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3779..4441)
                     /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(4442..4629)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(4776..4795)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4895..4917
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4955..4973)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5021..5524
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5554..5658
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5659..5727,5728..6519)
                     /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(5877..5896)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6690..7278
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7179..7198
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7432..7449
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7566..7587
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7602..7607,7608..7625,7626..7632)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7640..7656
                     /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     join(7645..7663,1..4)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
       61 aaaagctgga gctcttctca agcaaggttt tcagtataat gttacatgcg tacacgcgtc
      121 tgtacagaaa aaaaagaaaa atttgaaata taaataacgt tcttaatact aacataacta
      181 taaaaaaata aatagggacc tagacttcag gttgtctaac tccttccttt tcggttagag
      241 cggatgaatg cacgcgatcg tgcgatcttg agtgccagtg aatattaagt aatcatcaaa
      301 gtatatgtgt aattgtttat actcttagta aaggatgctc cctacaaggt ggctcttttc
      361 ttactaagcg cgttcagttt ccagccagcc gaaagaggga tatcagcttg agtgccagtg
      421 aatattaagt aatcatcaaa gtatatgtgt aattgtttat actcttagta aaggatgctc
      481 cctacaaggt ggctcttttc ttactaagcg cgttcagttt ccagccagcc gaaagaggga
      541 tatcagacct gcactcttga gtgccagtga atattaagta atcatcaaag tatatgtgta
      601 attgtttata ctcttagtaa aggatgctcc ctacaaggtg gctcttttct tactaagcgc
      661 gttcagtttc cagccagccg aaagagggat atcagcttga gtgccagtga atattaagta
      721 atcatcaaag tatatgtgta attgtttata ctcttagtaa aggatgctcc ctacaaggtg
      781 gctcttttct tactaagcgc gttcagtttc cagccagccg aaagagggat atcagatctg
      841 tcatcttgag tgccagtgaa tattaagtaa tcatcaaagt atatgtgtaa ttgtttatac
      901 tcttagtaaa ggatgctccc tacaaggtgg ctcttttctt actaagcgcg ttcagtttcc
      961 agccagccga aagagggata tcagtatata agaaagccat tcgggggatg aaaagctgac
     1021 aagagaataa cgaggaccag tttttatttg ttgtctagca agaattatac acgcacacat
     1081 acacgagagt ctacgatatc tttaaataac acatcaataa gtgcaggtaa aacaatgact
     1141 aagctacact ttgacactgc tgaaccagtc aagatcacac ttccaaatgg tttgacatac
     1201 gagcaaccaa ccggtctatt cattaacaac aagtttatga aagctcaaga cggtaagacc
     1261 tatcccgtcg aagatccttc cactgaaaac accgtttgtg aggtctcttc tgccaccact
     1321 gaagatgttg aatatgctat cgaatgtgcc gaccgtgctt tccacgacac tgaatgggct
     1381 acccaagacc caagagaaag aggccgtcta ctaagtaagt tggctgacga attggaaagc
     1441 caaattgact tggtttcttc cattgaagct ttggacaatg gtaaaacttt ggccttagcc
     1501 cgtggggatg ttaccattgc aatcaactgt ctaagagatg ctgctgccta tgccgacaaa
     1561 gtcaacggta gaacaatcaa caccggtgac ggctacatga acttcaccac cttagagcca
     1621 atcggtgtct gtggtcaaat tattccatgg aactttccaa taatgatgtt ggcttggaag
     1681 atcgccccag cattggccat gggtaacgtc tgtatcttga aacccgctgc tgtcacacct
     1741 ttaaatgccc tatactttgc ttctttatgt aagaaggttg gtattccagc tggtgtcgtc
     1801 aacatcgttc caggtcctgg tagaactgtt ggtgctgctt tgaccaacga cccaagaatc
     1861 agaaagctgg cttttaccgg ttctacagaa gtcggtaaga gtgttgctgt cgactcttct
     1921 gaatctaact tgaagaaaat cactttggaa ctaggtggta agtccgccca tttggtcttt
     1981 gacgatgcta acattaagaa gactttacca aatctagtaa acggtatttt caagaacgct
     2041 ggtcaaattt gttcctctgg ttctagaatt tacgttcaag aaggtattta cgacgaacta
     2101 ttggctgctt tcaaggctta cttggaaacc gaaatcaaag ttggtaatcc atttgacaag
     2161 gctaacttcc aaggtgctat cactaaccgt caacaattcg acacaattat gaactacatc
     2221 gatatcggta agaaagaagg cgccaagatc ttaactggtg gcgaaaaagt tggtgacaag
     2281 ggttacttca tcagaccaac cgttttctac gatgttaatg aagacatgag aattgttaag
     2341 gaagaaattt ttggaccagt tgtcactgtc gcaaagttca agactttaga agaaggtgtc
     2401 gaaatggcta acagctctga attcggtcta ggttctggta tcgaaacaga atctttgagc
     2461 acaggtttga aggtggccaa gatgttgaag gccggtaccg tctggatcaa cacatacaac
     2521 gattttgact ccagagttcc attcggtggt gttaagcaat ctggttacgg tagagaaatg
     2581 ggtgaagaag tctaccatgc atacactgaa gtaaaagctg tcagaattaa gttgtaaatc
     2641 gcacgcattc cgttggtaga tacgttgttg acacttctaa ataagcgaat ttcttatgat
     2701 ttatgatttt tattattaaa taagttataa aaaaaataag tgtatacaaa ttttaaagtg
     2761 actcttaggt tttaaaacga aaattcttat tcttgagtaa ctctttcctg taggtcaggt
     2821 tgctttctca ggtatagcat gaggtcgctc tcgagggggg gcccggtacc caattcgccc
     2881 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     2941 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     3001 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     3061 tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3121 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3181 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3241 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3301 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3361 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3421 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3481 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct
     3541 tacgcatctg tgcggtattt cacaccgcat agatccgtcg agttcaagag aaaaaaaaag
     3601 aaaaagcaaa aagaaaaaag gaaagcgcgc ctcgttcaga atgacacgta tagaatgatg
     3661 cattaccttg tcatcttcag tatcatactg ttcgtataca tacttactga cattcatagg
     3721 tatacatata tacacatgta tatatatcgt atgctgcagc tttaaataat cggtgtcact
     3781 acataagaac acctttggtg gagggaacat cgttggtacc attgggcgag gtggcttctc
     3841 ttatggcaac cgcaagagcc ttgaacgcac tctcactacg gtgatgatca ttcttgcctc
     3901 gcagacaatc aacgtggagg gtaattctgc tagcctctgc aaagctttca agaaaatgcg
     3961 ggatcatctc gcaagagaga tctcctactt tctccctttg caaaccaagt tcgacaactg
     4021 cgtacggcct gttcgaaaga tctaccaccg ctctggaaag tgcctcatcc aaaggcgcaa
     4081 atcctgatcc aaaccttttt actccacgca cggcccctag ggcctcttta aaagcttgac
     4141 cgagagcaat cccgcagtct tcagtggtgt gatggtcgtc tatgtgtaag tcaccaatgc
     4201 actcaacgat tagcgaccag ccggaatgct tggccagagc atgtatcata tggtccagaa
     4261 accctatacc tgtgtggacg ttaatcactt gcgattgtgt ggcctgttct gctactgctt
     4321 ctgcctcttt ttctgggaag atcgagtgct ctatcgctag gggaccaccc tttaaagaga
     4381 tcgcaatctg aatcttggtt tcatttgtaa tacgctttac tagggctttc tgctctgtca
     4441 tctttgcctt cgtttatctt gcctgctcat tttttagtat attcttcgaa gaaatcacat
     4501 tactttatat aatgtataat tcattatgtg ataatgccaa tcgctaagaa aaaaaaagag
     4561 tcatccgcta ggtggaaaaa aaaaaatgaa aatcattacc gaggcataaa aaaatataga
     4621 gtgtactaga ggaggccaag agtaatagaa aaagaaaatt gcgggaaagg actgtgttat
     4681 gacttccctg actaatgccg tgttcaaacg atacctggca gtgactccta gcgctcacca
     4741 agctcttaaa acgggaattt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     4801 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     4861 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     4921 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     4981 gttaatgtca tgataataat ggtttcttag ggtccttttc atcacgtgct ataaaaataa
     5041 ttataattta aattttttaa tataaatata taaattaaaa atagaaagta aaaaaagaaa
     5101 ttaaagaaaa aatagttttt gttttccgaa gatgtaaaag actctagggg gatcgccaac
     5161 aaatactacc ttttatcttg ctcttcctgc tctcaggtat taatgccgaa ttgtttcatc
     5221 ttgtctgtgt agaagaccac acacgaaaat cctgtgattt tacattttac ttatcgttaa
     5281 tcgaatgtat atctatttaa tctgcttttc ttgtctaata aatatatatg taaagtacgc
     5341 tttttgttga aattttttaa acctttgttt attttttttt cttcattccg taactcttct
     5401 accttcttta tttactttct aaaatccaaa tacaaaacat aaaaataaat aaacacagag
     5461 taaattccca aattattcca tcattaaaag atacgaggcg cgtgtaagtt acaggcaagc
     5521 gatccgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5581 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5641 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5701 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5761 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5821 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5881 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5941 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6001 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6061 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6121 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6181 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     6241 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6301 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6361 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6421 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6481 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6541 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6601 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6661 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6721 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6781 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6841 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6901 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6961 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7021 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7081 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7141 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7201 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7261 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7321 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     7381 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     7441 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     7501 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     7561 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     7621 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc aca
//
