LOCUS       sequence-430351-        5973 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integrative vector for constitutive expression of CDC19 at the
            TRP1 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5973)
  AUTHORS   Choi B, Tafur Rangel A, Kerkhoven EJ, Nygard Y
  TITLE     Engineering of Saccharomyces cerevisiae for enhanced metabolic 
            robustness and L-lactic acid production from lignocellulosic 
            biomass.
  JOURNAL   Metab Eng. 2024 Jul;84:23-33. doi: 10.1016/j.ymben.2024.05.003. Epub
            2024 May 23.
  PUBMED    38788894
REFERENCE   2  (bases 1 to 5973)
  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: "Metab 
            Eng."; date: "2024-07"; volume: "84:23-33. doi"; pages: " 
            10.1016/j.ymben.2024.05.003. Epub 2024 May 23"
COMMENT     Sequence Label: YIplac204-TDH3p-CDC19-CYC1t
FEATURES             Location/Qualifiers
     source          1..5973
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(142..147,148..165,166..172)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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     185..207
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     204..220
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        252..895
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     788..811
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     terminator      2464..2711
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2480..2498)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2718..2735)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2718..2734)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2727..2749)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2943..2962)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3062..3084
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3203..3877)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(3878..3979)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3981..3999)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4067..4171
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4172..4240,4241..5032)
                     /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(4390..4409)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5203..5791
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5692..5711
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5945..5962
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttca
      241 gttcgagttt atcattatca atactgccat ttcaaagaat acgtaaataa ttaatagtag
      301 tgattttcct aactttattt agtcaaaaaa ttagcctttt aattctgctg taacccgtac
      361 atgcccaaaa tagggggcgg gttacacaga atatataaca tcgtaggtgt ctgggtgaac
      421 agtttattcc tggcatccac taaatataat ggagcccgct ttttaagctg gcatccagaa
      481 aaaaaaagaa tcccagcacc aaaatattgt tttcttcacc aaccatcagt tcataggtcc
      541 attctcttag cgcaactaca gagaacaggg gcacaaacag gcaaaaaacg ggcacaacct
      601 caatggagtg atgcaacctg cctggagtaa atgatgacac aaggcaattg acccacgcat
      661 gtatctatct cattttctta caccttctat taccttctgc tctctctgat ttggaaaaag
      721 ctgaaaaaaa aggttgaaac cagttccctg aaattattcc cctacttgac taataagtat
      781 ataaagacgg taggtattga ttgtaattct gtaaatctat ttcttaaact tcttaaattc
      841 tacttttata gttagtcttt tttttagttt taaaacacca agaacttagt ttcgaataaa
      901 cacacctgca ggtcgacatg tctagattag aaagattgac ctcattaaac gttgttgctg
      961 gttctgactt gagaagaacc tccatcattg gtaccatcgg tccaaagacc aacaacccag
     1021 aaaccttggt tgctttgaga aaggctggtt tgaacattgt ccgtatgaac ttctctcacg
     1081 gttcttacga ataccacaag tctgtcattg acaacgccag aaagtccgaa gaattgtacc
     1141 caggtagacc attggccatt gctttggaca ccaagggtcc agaaatcaga actggtacca
     1201 ccaccaacga tgttgactac ccaatcccac caaaccacga aatgatcttc accaccgatg
     1261 acaagtacgc taaggcttgt gacgacaaga tcatgtacgt tgactacaag aacatcacca
     1321 aggtcatctc cgctggtaga atcatctacg ttgatgatgg tgttttgtct ttccaagttt
     1381 tggaagtcgt tgacgacaag actttgaagg tcaaggcttt gaacgccggt aagatctgtt
     1441 cccacaaggg tgtcaactta ccaggtaccg atgtcgattt gccagctttg tctgaaaagg
     1501 acaaggaaga tttgagattc ggtgtcaaga acggtgtcca catggtcttc gcttctttca
     1561 tcagaaccgc caacgatgtt ttgaccatca gagaagtctt gggtgaacaa ggtaaggacg
     1621 tcaagatcat tgtcaagatt gaaaaccaac aaggtgttaa caacttcgac gaaatcttga
     1681 aggtcactga cggtgttatg gttgccagag gtgacttggg tattgaaatc ccagccccag
     1741 aagtcttggc tgtccaaaag aaattgattg ctaagtctaa cttggctggt aagccagtta
     1801 tctgtgctac ccaaatgttg gaatccatga cttacaaccc aagaccaacc agagctgaag
     1861 tttccgatgt cggtaacgct atcttggatg gtgctgactg tgttatgttg tctggtgaaa
     1921 ccgccaaggg taactaccca atcaacgccg ttaccactat ggctgaaacc gctgtcattg
     1981 ctgaacaagc tatcgcttac ttgccaaact acgatgacat gagaaactgt actccaaagc
     2041 caacctccac caccgaaacc gtcgctgcct ccgctgtcgc tgctgttttc gaacaaaagg
     2101 ccaaggctat cattgtcttg tccacttccg gtaccacccc aagattggtt tccaagtaca
     2161 gaccaaactg tccaatcatc ttggttacca gatgcccaag agctgctaga ttctctcact
     2221 tgtacagagg tgtcttccca ttcgttttcg aaaaggaacc tgtctctgac tggactgatg
     2281 atgttgaagc ccgtatcaac ttcggtattg aaaaggctaa ggaattcggt atcttgaaga
     2341 agggtgacac ttacgtttcc atccaaggtt tcaaggccgg tgctggtcac tccaacactt
     2401 tgcaagtctc taccgtttaa ggatccccgg gtaccgagct cggccccttt tcctttgtcg
     2461 atatcatgta attagttatg tcacgcttac attcacgccc tccccccaca tccgctctaa
     2521 ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt tttatagtta
     2581 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacaaacgcg
     2641 tgtacgcatg taacattata ctgaaaacct tgcttgagaa ggttttggga cgctcgaagg
     2701 ctttaatttg cgaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     2761 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     2821 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
     2881 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     2941 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     3001 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     3061 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     3121 ggcgatcttt tatgcttgct tttcaaaagg cttgcaggca agtgcacaaa caatacttaa
     3181 ataaatacta ctcagtaata acctatttct tagcattttt gacgaaattt gctattttgt
     3241 tagagtcttt tacaccattt gtctccacac ctccgcttac atcaacacca ataacgccat
     3301 ttaatctaag cgcatcacca acattttctg gcgtcagtcc accagctaac ataaaatgta
     3361 agctctcggg gctctcttgc cttccaaccc agtcagaaat cgagttccaa tccaaaagtt
     3421 cacctgtccc acctgcttct gaatcaaaca agggaataaa cgaatgaggt ttctgtgaag
     3481 ctgcactgag tagtatgttg cagtcttttg gaaatacgag tcttttaata actggcaaac
     3541 cgaggaactc ttggtattct tgccacgact catctccatg cagttggacg atatcaatgc
     3601 cgtaatcatt gaccagagcc aaaacatcct ccttaggttg attacgaaac acgccaacca
     3661 agtatttcgg agtgcctgaa ctatttttat atgcttttac aagacttgaa attttccttg
     3721 caataaccgg gtcaattgtt ctctttctat tgggcacaca tataataccc agcaagtcag
     3781 catcggaatc tagtgcacat tctgcggcct ctgtgctctg caagccgcaa actttcacca
     3841 atggaccaga actacctgtg aaattaataa cagacatact ccaagctgcc tttgtgtgct
     3901 taatcacgta tactcacgtg ctcaatagtc accaatgccc tccctcttgg ccctctcctt
     3961 ttcttttttc gaccgaattg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     4021 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4081 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4141 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4201 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4261 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     4321 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     4381 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     4441 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     4501 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     4561 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     4621 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     4681 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     4741 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     4801 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     4861 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     4921 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     4981 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5041 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5101 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5161 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5221 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5281 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5341 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5401 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5461 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5521 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5581 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5641 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5701 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5761 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5821 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5881 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     5941 gcgcagcgag tcagtgagcg aggaagcgga aga
//
