LOCUS       sequence-177609-        8258 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression in S.cerevisiae of FjTAL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8258)
  AUTHORS   Eudes A, Mouille M, Robinson DS, Benites VT, Wang G, Roux L, Tsai 
            YL, Baidoo EE, Chiu TY, Heazlewood JL, Scheller HV, Mukhopadhyay A, 
            Keasling JD, Deutsch S, Loque D
  TITLE     Exploiting members of the BAHD acyltransferase family to synthesize 
            multiple hydroxycinnamate and benzoate conjugates in yeast.
  JOURNAL   Microb Cell Fact. 2016 Nov 21;15(1):198.
  PUBMED    27871334
REFERENCE   2  (bases 1 to 8258)
  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"; journalName: "Microb Cell
            Fact."; date: "2016-11-21"; volume: "15(1)"; pages: "198"
COMMENT     Sequence Label: pRS423-FjTAL(JBEI-15466)
FEATURES             Location/Qualifiers
     source          1..8258
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(158..746)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(238..257)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(917..1708,1709..1777))
                     /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     1540..1559
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1778..1882)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1909..3251
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(2872..2919)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     3292..3310
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3348..3370)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3470..3489
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3636..3823
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             3824..4486
                     /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"
     rep_origin      complement(4746..5201)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4883..4904)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5095..5114
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5327..5349
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5341..5358
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5342..5358
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5368..5387
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5368..5386
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     5411..5427
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     5412..5428
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      complement(5437..5684)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     5650..5668
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     protein_bind    5713..5737
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(7262..7286)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(7300..7942)
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     complement(7383..7406)
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     complement(7975..7995)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7975..7993)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8014..8030)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8014..8030)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8027..8049)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8038..8054
                     /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)."
     promoter        complement(join(8062..8068,8069..8086,8087..8092))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8107..8128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(join(8245..8258,1..4))
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
       61 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
      121 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
      181 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
      241 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
      301 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
      361 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
      421 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
      481 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
      541 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
      601 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
      661 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
      721 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
      781 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
      841 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
      901 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
      961 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     1021 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     1081 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     1141 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     1201 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     1261 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     1321 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     1381 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     1441 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     1501 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     1561 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     1621 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     1681 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     1741 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     1801 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     1861 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgaa cgaagcatct
     1921 gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat
     1981 ctgagctgca tttttacaga acagaaatgc aacgcgaaag cgctatttta ccaacgaaga
     2041 atctgtgctt catttttgta aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa
     2101 agaatctgag ctgcattttt acagaacaga aatgcaacgc gagagcgcta ttttaccaac
     2161 aaagaatcta tacttctttt ttgttctaca aaaatgcatc ccgagagcgc tatttttcta
     2221 acaaagcatc ttagattact ttttttctcc tttgtgcgct ctataatgca gtctcttgat
     2281 aactttttgc actgtaggtc cgttaaggtt agaagaaggc tactttggtg tctattttct
     2341 cttccataaa aaaagcctga ctccacttcc cgcgtttact gattactagc gaagctgcgg
     2401 gtgcattttt tcaagataaa ggcatccccg attatattct ataccgatgt ggattgcgca
     2461 tactttgtga acagaaagtg atagcgttga tgattcttca ttggtcagaa aattatgaac
     2521 ggtttcttct attttgtctc tatatactac gtataggaaa tgtttacatt ttcgtattgt
     2581 tttcgattca ctctatgaat agttcttact acaatttttt tgtctaaaga gtaatactag
     2641 agataaacat aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg
     2701 atgggtaggt tatataggga tatagcacag agatatatag caaagagata cttttgagca
     2761 atgtttgtgg aagcggtatt cgcaatattt tagtagctcg ttacagtccg gtgcgttttt
     2821 ggttttttga aagtgcgtct tcagagcgct tttggttttc aaaagcgctc tgaagttcct
     2881 atactttcta gagaatagga acttcggaat aggaacttca aagcgtttcc gaaaacgagc
     2941 gcttccgaaa atgcaacgcg agctgcgcac atacagctca ctgttcacgt cgcacctata
     3001 tctgcgtgtt gcctgtatat atatatacat gagaagaacg gcatagtgcg tgtttatgct
     3061 taaatgcgta cttatatgcg tctatttatg taggatgaaa ggtagtctag tacctcctgt
     3121 gatattatcc cattccatgc ggggtatcgt atgcttcctt cagcactacc ctttagctgt
     3181 tctatatgct gccactcctc aattggatta gtctcatcct tcaatgctat catttccttt
     3241 gatattggat catactaaga aaccattatt atcatgacat taacctataa aaataggcgt
     3301 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     3361 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     3421 cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag
     3481 cagattgtac tgagagtgca ccataaattc ccgttttaag agcttggtga gcgctaggag
     3541 tcactgccag gtatcgtttg aacacggcat tagtcaggga agtcataaca cagtcctttc
     3601 ccgcaatttt ctttttctat tactcttggc ctcctctagt acactctata tttttttatg
     3661 cctcggtaat gattttcatt tttttttttc cacctagcgg atgactcttt ttttttctta
     3721 gcgattggca ttatcacata atgaattata cattatataa agtaatgtga tttcttcgaa
     3781 gaatatacta aaaaatgagc aggcaagata aacgaaggca aagatgacag agcagaaagc
     3841 cctagtaaag cgtattacaa atgaaaccaa gattcagatt gcgatctctt taaagggtgg
     3901 tcccctagcg atagagcact cgatcttccc agaaaaagag gcagaagcag tagcagaaca
     3961 ggccacacaa tcgcaagtga ttaacgtcca cacaggtata gggtttctgg accatatgat
     4021 acatgctctg gccaagcatt ccggctggtc gctaatcgtt gagtgcattg gtgacttaca
     4081 catagacgac catcacacca ctgaagactg cgggattgct ctcggtcaag cttttaaaga
     4141 ggccctaggg gccgtgcgtg gagtaaaaag gtttggatca ggatttgcgc ctttggatga
     4201 ggcactttcc agagcggtgg tagatctttc gaacaggccg tacgcagttg tcgaacttgg
     4261 tttgcaaagg gagaaagtag gagatctctc ttgcgagatg atcccgcatt ttcttgaaag
     4321 ctttgcagag gctagcagaa ttaccctcca cgttgattgt ctgcgaggca agaatgatca
     4381 tcaccgtagt gagagtgcgt tcaaggctct tgcggttgcc ataagagaag ccacctcgcc
     4441 caatggtacc aacgatgttc cctccaccaa aggtgttctt atgtagtgac accgattatt
     4501 taaagctgca gcatacgata tatatacatg tgtatatatg tatacctatg aatgtcagta
     4561 agtatgtata cgaacagtat gatactgaag atgacaaggt aatgcatcat tctatacgtg
     4621 tcattctgaa cgaggcgcgc tttccttttt tctttttgct ttttcttttt ttttctcttg
     4681 aactcgacgg atctatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     4741 tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     4801 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     4861 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     4921 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     4981 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     5041 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     5101 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     5161 caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg
     5221 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     5281 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     5341 tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga attgggtacc
     5401 gggccccccc tcgaggtcga cggtatcgat aagcttgcaa attaaagcct tcgagcgtcc
     5461 caaaaccttc tcaagcaagg ttttcagtat aatgttacat gcgtacacgc gtctgtacag
     5521 aaaaaaaaga aaaatttgaa atataaataa cgttcttaat actaacataa ctataaaaaa
     5581 ataaataggg acctagactt caggttgtct aactccttcc ttttcggtta gagcggatgt
     5641 ggggggaggg cgtgaatgta agcgtgacat aactaattac atgatgactc tagcgaattc
     5701 tcaattaact acaccacttt gtacaagaaa gctgggtctc aattgttgat aaggtggtct
     5761 ttcactttct gcacgaatgg gtacataact tggtcctctt tgaatgtagg aataatattt
     5821 ctaatttcat cgtaccactt tttagaaact gaagaaatct tatccttttg acccaagtaa
     5881 tcgatagctt gaacgatagt gatcatttcg atagcaagaa cttcgaaagc gttttcaata
     5941 actttagatg taataacagc tgagttagta cccatagaaa caatatcttg gttatcattg
     6001 ttattaggga tagaatgaac atacattggg tttgacaaca tttgagattc agctgtagtt
     6061 gatgtagcag tgaattgaac tccttgcata ccgaaattaa aaccaagtgt acccaaatta
     6121 acgaaaggtg gaagaagttc gttaatttta gaatttaaaa ggtagttaag ttgtctttca
     6181 gccaacattg taagcttagt gataacgatc ttcaacttat ccatctcaag tgagatgtaa
     6241 tctccatgga agtttccacc atgataaaca tgttgattct taacatcaat aattgggtta
     6301 tcgttagcag aattaaactc atcttccaag attgaagcaa cgttattaat tgtttcaaga
     6361 acaggtccta aaatttgtgg aacacatcta agagagtagt attcttgaac cttttctttg
     6421 aaaatttctt ctgtgttttc acctgaatac aaatgatctt ctctctttct gattaaagtt
     6481 gaatcagaca aattttgtct catcttaaga gcgatttctt gttgaccctt atgtctctta
     6541 gtttggttca attcagcaga gaaatgatca tcgtaagctt gaacaagttc gttaatagca
     6601 catgaagatt ttaatgacca atcaagcaac tttttagcat ggtaaacgtt aacaactcca
     6661 atacctgtca taacagaagt accattaatc aaagcaagac cttctctgat ttcaacttgg
     6721 ataggcttta atccttcaat ttcgaaaact tcaggtgttg gccttctttc acccttatag
     6781 aaaacttctc cttcaccgat aagaaccaaa gcaagatgag aaagttgaac caaatctcct
     6841 gaagcaccaa ctccaccatg ttcgaagatt aaaggagtga tatccttgtt aatcaattca
     6901 gacataagat taataactga tggatgaact cctgagttac ccaaagacaa tgtattaagt
     6961 ctagctaaga tagcagcctt agcacaaaca ggagacaatg gttttccagt acctgaagaa
     7021 tgtgatctga tcaaattgta ttgcaattgg atttgatcag attctttaat tctgtattga
     7081 gccattggtc cgaaaccagt gttaactcca tagataacct tattaccaga aaattccttc
     7141 aagaaattaa aagattcgtt aactctgtta acaaccacgt ctgagattgt aactttctga
     7201 tttccgaata taatagcttc aaactcctct aaactaaggt actcgttgat agtgttcatg
     7261 aagcctgctt ttttgtacaa acttgtggga tccgctagct cgaaactaag ttctggtgtt
     7321 ttaaaactaa aaaaaagact aactataaaa gtagaattta agaagtttaa gaaatagatt
     7381 tacagaatta caatcaatac ctaccgtctt tatatactta ttagtcaagt aggggaataa
     7441 tttcagggaa ctggtttcaa cctttttttt cagctttttc caaatcagag agagcagaag
     7501 gtaatagaag gtgtaagaaa atgagataga tacatgcgtg ggtcaattgc cttgtgtcat
     7561 catttactcc aggcaggttg catcactcca ttgaggttgt gcccgttttt tgcctgtttg
     7621 tgcccctgtt ctctgtagtt gcgctaagag aatggaccta tgaactgatg gttggtgaag
     7681 aaaacaatat tttggtgctg ggattctttt tttttctgga tgccagctta aaaagcgggc
     7741 tccattatat ttagtggatg ccaggaataa actgttcacc cagacaccta cgatgttata
     7801 tattctgtgt aacccgcccc ctattttggg catgtacggg ttacagcaga attaaaaggc
     7861 taattttttg actaaataaa gttaggaaaa tcactactat taattattta cgtattcttt
     7921 gaaatggcag tattgataat gataaactcc gcggtggagc tccagctttt gttcccttta
     7981 gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     8041 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     8101 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     8161 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     8221 gcgtattggg cgctcttccg cttcctcgct cactgact
//
