LOCUS       sequence-76066-e        6555 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of the human TPI Ile170Thr allele in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6555)
  AUTHORS   Gruning NM, Du D, Keller MA, Luisi BF, Ralser M
  TITLE     Inhibition of triosephosphate isomerase by phosphoenolpyruvate in 
            the feedback-regulation of glycolysis.
  JOURNAL   Open Biol. 2014 Mar 5;4(3):130232. doi: 10.1098/rsob.130232.
  PUBMED    24598263
REFERENCE   2  (bases 1 to 6555)
  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: "Open 
            Biol."; date: "2014-03-5"; volume: "4(3):130232. doi"; pages: " 
            10.1098/rsob.130232"
COMMENT     Sequence Label: p413GPD-hTPI Ile170Thr
FEATURES             Location/Qualifiers
     source          1..6555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    70..573
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        610..714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(715..783,784..1575)
                     /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(933..952)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1746..2334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2235..2254
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2488..2505
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2658..2663,2664..2681,2682..2688)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2696..2712
                     /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     2701..2723
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2720..2736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2720..2736
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2755..2775
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2757..2775
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2800..3443
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     3337..3360
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     3449..3465
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3449..3465
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      4223..4470
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4239..4257)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4488..4507)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4489..4507)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4517..4534)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4517..4533)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4526..4548)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4678..5133
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4765..4784)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4975..4996
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5393..6052)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(6053..6239)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(6386..6405)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6505..6527
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata
      121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
      181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
      241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
      301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct
      361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
      421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
      481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
      541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg
      601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1141 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1501 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2101 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2221 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact
     2641 cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg
     2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     2761 aaccctcact aaagggaaca aaagctggag ctcagtttat cattatcaat actcgccatt
     2821 tcaaagaata cgtaaataat taatagtagt gattttccta actttattta gtcaaaaaat
     2881 tagcctttta attctgctgt aacccgtaca tgcccaaaat agggggcggg ttacacagaa
     2941 tatataacat cgtaggtgtc tgggtgaaca gtttattcct ggcatccact aaatataatg
     3001 gagcccgctt tttaagctgg catccagaaa aaaaaagaat cccagcacca aaatattgtt
     3061 ttcttcacca accatcagtt cataggtcca ttctcttagc gcaactacag agaacagggg
     3121 cacaaacagg caaaaaacgg gcacaacctc aatggagtga tgcaacctgc ctggagtaaa
     3181 tgatgacaca aggcaattga cccacgcatg tatctatctc attttcttac accttctatt
     3241 accttctgct ctctctgatt tggaaaaagc tgaaaaaaaa ggttgaaacc agttccctga
     3301 aattattccc ctacttgact aataagtata taaagacggt aggtattgat tgtaattctg
     3361 taaatctatt tcttaaactt cttaaattct acttttatag ttagtctttt ttttagtttt
     3421 aaaacaccag aacttagttt cgacggattc tagaactagt ggatccatgg cgccctccag
     3481 gaagttcttc gttgggggaa actggaagat gaacgggcgg aagcagagtc tgggggagct
     3541 catcggcact ctgaacgcgg ccaaggtgcc ggccgacacc gaggtggttt gtgctccccc
     3601 tactgcctat atcgacttcg cccggcagaa gctagatccc aagattgctg tggctgcgca
     3661 gaactgctac aaagtgacta atggggcttt tactggggag atcagccctg gcatgatcaa
     3721 agactgcgga gccacgtggg tggtcctggg gcactcagag agaaggcatg tctttgggga
     3781 gtcagatgag ctgattgggc agaaagtggc ccatgctctg gcagagggac tcggagtaat
     3841 cgcctgcatt ggggagaagc tagatgaaag ggaagctggc atcactgaga aggttgtttt
     3901 cgagcagaca aaggtcatcg cagataacgt gaaggactgg agcaaggtcg tcctggccta
     3961 tgagcctgtg tgggccactg gtactggcaa gactgcaaca ccccaacagg cccaggaagt
     4021 acacgagaag ctccgaggat ggctgaagtc caacgtctct gatgcggtgg ctcagagcac
     4081 ccgtatcatt tatggaggct ctgtgactgg ggcaacctgc aaggagctgg ccagccagcc
     4141 tgatgtggat ggcttccttg tgggtggtgc ttccctcaag cccgaattcg tggacatcat
     4201 caatgccaaa caatgactcg agtcatgtaa ttagttatgt cacgcttaca ttcacgccct
     4261 ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct
     4321 atttattttt ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt
     4381 ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag
     4441 gttttgggac gctcgaaggc tttaatttgc ggccggtacc caattcgccc tatagtgagt
     4501 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     4561 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     4621 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcgacg
     4681 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4741 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4801 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4861 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4921 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4981 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     5041 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     5101 cgaattttaa caaaatatta acgtttacaa tttcctgatg cggtattttc tccttacgca
     5161 tctgtgcggt atttcacacc gcatagatcc gtcgagttca agagaaaaaa aaagaaaaag
     5221 caaaaagaaa aaaggaaagc gcgcctcgtt cagaatgaca cgtatagaat gatgcattac
     5281 cttgtcatct tcagtatcat actgttcgta tacatactta ctgacattca taggtataca
     5341 tatatacaca tgtatatata tcgtatgctg cagctttaaa taatcggtgt cactacataa
     5401 gaacaccttt ggtggaggga acatcgttgg taccattggg cgaggtggct tctcttatgg
     5461 caaccgcaag agccttgaac gcactctcac tacggtgatg atcattcttg cctcgcagac
     5521 aatcaacgtg gagggtaatt ctgctagcct ctgcaaagct ttcaagaaaa tgcgggatca
     5581 tctcgcaaga gagatctcct actttctccc tttgcaaacc aagttcgaca actgcgtacg
     5641 gcctgttcga aagatctacc accgctctgg aaagtgcctc atccaaaggc gcaaatcctg
     5701 atccaaacct ttttactcca cgcgccagta gggcctcttt aaaagcttga ccgagagcaa
     5761 tcccgcagtc ttcagtggtg tgatggtcgt ctatgtgtaa gtcaccaatg cactcaacga
     5821 ttagcgacca gccggaatgc ttggccagag catgtatcat atggtccaga aaccctatac
     5881 ctgtgtggac gttaatcact tgcgattgtg tggcctgttc tgctactgct tctgcctctt
     5941 tttctgggaa gatcgagtgc tctatcgcta ggggaccacc ctttaaagag atcgcaatct
     6001 gaatcttggt ttcatttgta atacgcttta ctagggcttt ctgctctgtc atctttgcct
     6061 tcgtttatct tgcctgctca ttttttagta tattcttcga agaaatcaca ttactttata
     6121 taatgtataa ttcattatgt gataatgcca atcgctaaga aaaaaaaaga gtcatccgct
     6181 aggggaaaaa aaaaaatgaa aatcattacc gaggcataaa aaaatataga gtgtactaga
     6241 ggaggccaag agtaatagaa aaagaaaatt gcgggaaagg actgtgttat gacttccctg
     6301 actaatgccg tgttcaaacg atacctggca gtgactccta gcgctcacca agctcttaaa
     6361 acgggaattt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     6421 cccgacaccc gccaacacgc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     6481 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     6541 caccgaaacg cgcga
//
