LOCUS       sequence-254153-        5093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast pathway position 7. ACDH transcription unit with the TDH3 
            promoter and RPL3 terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5093)
  AUTHORS   Young EM, Zhao Z, Gielesen BEM, Wu L, Benjamin Gordon D, Roubos JA, 
            Voigt CA
  TITLE     Iterative algorithm-guided design of massive strain libraries, 
            applied to itaconic acid production in yeast.
  JOURNAL   Metab Eng. 2018 Jul;48:33-43. doi: 10.1016/j.ymben.2018.05.002. Epub
            2018 May 9.
  PUBMED    29753070
REFERENCE   2  (bases 1 to 5093)
  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: "2018-07"; volume: "48:33-43. doi"; pages: " 
            10.1016/j.ymben.2018.05.002. Epub 2018 May 9"
COMMENT     Sequence Label: pDSM-f3-Ptdh3-ACDH-Trpl3
FEATURES             Location/Qualifiers
     source          1..5093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    113..146
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     promoter        522..809
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     679..702
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     protein_bind    complement(2513..2546)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(3016..3033)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3016..3032)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3131..3148)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3302..3890)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3382..3401)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4012..4821)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(4822..4913)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4980..4999
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 catgcgagag tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg
       61 ggcctttcgc ccgggctaat tagggggtgt cgcccttatt cgactctata gtgaagttcc
      121 tattctctag aaagtatagg aacttctgaa gtggggaaac cttcgaatcc agccagcatg
      181 tcgacaccca caagatgtag tgcacgtgct tagtcaaaaa attagccttt taattctgct
      241 gtaacccgta catgcccaaa atagggggcg ggttacacag aatatataac atcgtaggtg
      301 tctgggtgaa cagtttattc ctggcatcca ctaaatataa tggagcccgc tttttaagct
      361 ggcatccaga aaaaaaaaga atcccagcac caaaatattg ttttcttcac caaccatcag
      421 ttcataggtc cattctctta gcgcaactac agagaacagg ggcacaaaca ggcaaaaaac
      481 gggcacaacc tcaatggagt gatgcaacct gcctggagta aatgatgaca caaggcaatt
      541 gacccacgca tgtatctatc tcattttctt acaccttcta ttaccttctg ctctctctga
      601 tttggaaaaa gctgaaaaaa aaggttgaaa ccagttccct gaaattattc ccctacttga
      661 ctaataagta tataaagacg gtaggtattg attgtaattc tgtaaatcta tttcttaaac
      721 ttcttaaatt ctacttttat agttagtctt ttttttagtt ttaaaacacc aagaacttag
      781 tttcgaataa acacacataa acaaacaaaa tggaatcttt ggaattggaa caattagtca
      841 agaaggtttt gttggaaaaa ttggctgaac aaaaggaagt tccaaccaag accaccaccc
      901 aaggtgccaa gtccggtgtt ttcgacaccg tcgatgaagc tgtccaagct gctgtcattg
      961 ctcaaaactg ttacaaggaa aaatctttgg aagaaagaag aaacgttgtc aaggccatca
     1021 gagaagcttt gtacccagaa atcgaaacca ttgccaccag agctgttgct gaaaccggta
     1081 tgggtaatgt cactgacaag atcttgaaga acactttggc catcgaaaag accccaggtg
     1141 ttgaagattt gtacactgaa gttgccactg gtgacaacgg tatgactttg tacgaattgt
     1201 ctccatacgg tgtcatcggt gctgttgccc catctaccaa cccaactgaa actttgatct
     1261 gtaactccat tggtatgttg gctgctggta atgctgtttt ctactctcct cacccaggtg
     1321 ccaagaacat ctctttatgg ttaatcgaaa aattgaacac cattgtccgt gactcttgtg
     1381 gtatcgacaa cttgattgtc actgttgcca agccttccat ccaagctgct caagaaatga
     1441 tgaaccatcc aaaggtccca ttgttggtta tcactggtgg tccaggtgtt gtcttgcaag
     1501 ctatgcaatc tggtaagaag gtcattggtg ctggtgctgg taacccacca tctatcgtcg
     1561 atgaaactgc taacattgaa aaggctgccg ctgatatcgt tgacggtgct tctttcgacc
     1621 acaacatcct atgtattgct gaaaaatccg ttgttgccgt tgactccatt gctgatttct
     1681 tattattcca aatggaaaag aacggtgctt tgcacgttac caacccatct gatatccaaa
     1741 aattggaaaa ggttgctgtc actgacaagg gtgtcaccaa caagaaattg gttggtaagt
     1801 ctgctactga aatcttgaag gaagctggta ttgcttgtga cttcactcca agattaatca
     1861 ttgtcgaaac tgaaaagtcc cacccatttg ccaccgttga attgttgatg ccaattgtcc
     1921 cagttgtcag agttccagac ttcgatgaag ctttggaagt tgccatcgaa ttggaacaag
     1981 gtttgcacca cactgctacc atgcactctc aaaacatctc cagattgaac aaggctgcta
     2041 gagacatgca aacttccatc tttgtcaaga acggtccatc tttcgctggt ttaggtttca
     2101 gaggtgaagg ttccactact ttcaccattg ctaccccaac tggtgaaggt accaccaccg
     2161 ctagacattt cgctagaaga agaagatgtg ttttgactga tggtttctcc atacgttaaa
     2221 aagttttgtt agaaaataaa tcatttttta attgagcatt cttattccta ttttatttaa
     2281 atagttttat gtattgttag ctacatacaa cagtttaaat caaattttct ttttcccaag
     2341 tccaaaatgg aggtttattt tgatgacccg catgcgatta tgttttgaaa gtataagact
     2401 acatacatgt acatatattt aaacatgtaa acccgtccat tatattgccc tcagaaagcc
     2461 tgtatgcgaa gccacaatcc tttccaacag accatactaa gtaaaatgaa gtgaagttcc
     2521 tatactttct agagaatagg aacttctata gtgagtcgaa taagggcgac acaaaattta
     2581 ttctaaatgc ataataaata ctgataacat cttatagttt gtattatatt ttgtattatc
     2641 gttgacatgt ataattttga tatcaaaaac tgattttccc tttattattt tcgagattta
     2701 ttttcttaat tctctttaac aaactagaaa tattgtatat acaaaaaatc ataaataata
     2761 gatgaatagt ttaattatag gtgttcatca atcgaaaaag caacgtatct tatttaaagt
     2821 gcgttgcttt tttctcattt ataaggttaa ataattctca tatatcaagc aaagtgacag
     2881 gcgcccttaa atattctgac aaatgctctt tccctaaact ccccccataa aaaaacccgc
     2941 cgaagcgggt ttttacgtta tttgcggatt aacgattact cgttatcaga accgcccagg
     3001 gggcccgagc ttaagactgg ccgtcgtttt acaacacaga aagagtttgt agaaacgcaa
     3061 aaaggccatc cgtcaggggc cttctgctta gtttgatgcc tggcagttcc ctactctcgc
     3121 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3181 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3241 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3301 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3361 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3421 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3481 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3541 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3601 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3661 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggg
     3721 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     3781 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3841 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3901 tgatcttttc tacggggtct gacgctcagt ggaacgacgc gcgcgtaact cacgttaagg
     3961 gattttggtc atgagcttgc gccgtcccgt caagtcagcg taatgctctg cttagaaaaa
     4021 ctcatcgagc atcaaatgaa attgcaattt attcatatca ggattatcaa taccatattt
     4081 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
     4141 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
     4201 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
     4261 tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
     4321 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
     4381 gaggcgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg agtgcaaccg
     4441 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
     4501 tacctggaac gctgtttttc cggggatcgc agtggtgagt aaccatgcat catcaggagt
     4561 acggataaaa tgcttgatgg tcggaagtgg cataaattcc gtcagccagt ttagtctgac
     4621 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
     4681 cgcatcgggc ttcccataca agcgatagat tgtcgcacct gattgcccga cattatcgcg
     4741 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctcgacgt
     4801 ttcccgttga atatggctca tattcttcct ttttcaatat tattgaagca tttatcaggg
     4861 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4921 cagtgttaca accaattaac caattctgaa cattatcgcg agcccattta tacctgaata
     4981 tggctcataa caccccttgt ttgcctggcg gcagtagcgc ggtggtccca cctgacccca
     5041 tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggact ccc
//
