LOCUS       sequence-254061-        6045 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast pathway position 4. ACO transcription unit with the FBA1 
            promoter and GPM1 terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6045)
  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 6045)
  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-cd-Pfba1p-ACO-Tgpm1
FEATURES             Location/Qualifiers
     source          1..6045
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     103..120
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     104..120
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    590..623
                     /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)"
     protein_bind    complement(3942..3975)
                     /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(4182..4201)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        4268..4359
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4360..5169
                     /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"
     rep_origin      5291..5879
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5780..5799
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(6033..6045,1..5)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcgaggaagc ggaaggcgag agtagggaac tgccaggcat caaactaagc agaaggcccc
       61 tgacggatgg cctttttgcg tttctacaaa ctctttctgt gttgtaaaac gacggccagt
      121 cttaagctcg ggccccctgg gcggttctga taacgagtaa tcgttaatcc gcaaataacg
      181 taaaaacccg cttcggcggg tttttttatg gggggagttt agggaaagag catttgtcag
      241 aatatttaag ggcgcctgtc actttgcttg atatatgaga attatttaac cttataaatg
      301 agaaaaaagc aacgcacttt aaataagata cgttgctttt tcgattgatg aacacctata
      361 attaaactat tcatctatta tttatgattt tttgtatata caatatttct agtttgttaa
      421 agagaattaa gaaaataaat ctcgaaaata ataaagggaa aatcagtttt tgatatcaaa
      481 attatacatg tcaacgataa tacaaaatat aatacaaact ataagatgtt atcagtattt
      541 attatgcatt tagaataaat tttgtgtcgc ccttattcga ctcactatag aagttcctat
      601 tctctagaaa gtataggaac ttcacttcat tttgcggaat attggcggaa cggacacacg
      661 tggatacaaa cctggacaac gttgaggtat tgctataaca tgtcatgtca ccattaatta
      721 ccactcattc caacatactg tccctgacga aaactgttta tgcgagcgca aaatgaacct
      781 attcaattaa taatactcct caaattgcta ccacgacagg ctgaaaaaag atgtcgacga
      841 ataatgtaat cgtgtcaaac aaacgttatc ttgactatat atagctactt aatttgaaat
      901 aataaactat acaagtctat atcattgata tattcagtaa gaaaaatgga gggaaaaaga
      961 aatcatcaaa tcattcattc ttcagacttt acttcttttc atcagccttg atcttgttca
     1021 aagcagaacc gtatttgaac cattcgattt gttcatcgtt gaaagtgtga gtcaagacag
     1081 catcccatgg cttaccgttc tttgggtgaa ctctcatggt gactggctta cctggagcca
     1141 attcagccaa acctaagatg tcaattctgt catctgggtt gatcttgtcg tagtcggctg
     1201 ggttcttgaa gttcaatggc aatagacctt gcttcttcaa gttggtttcg tgaatacgag
     1261 cgaaagattt ggtaataata gcgaaaccac ccaagaatct tggttccaaa gcagcgtgtt
     1321 cacgagaaga accttcaccg aagttttcgt caccgatgac aacccatttg ataccttgat
     1381 ctctgtagtc tctggcagtg tctggaacac ccttgtattc accagtgtaa acgttcttga
     1441 cacagttagc cttcttgttt tcggcattaa tggcaccaat catgtagttg ttggagatgt
     1501 tttccaagtg acctctgtat ttcaaccatg gaccagccat ggaaatgtgg tcggtggtag
     1561 tcttaccaac agccttgatt aagattggca tgtccttagc gtccttacca tcccatggct
     1621 tgaaaggttt caacaattgt aatctgtcag aagttggaga aaccttgact tcgacggtgg
     1681 atctgtcggc tggtggagct tggtaggtgt tttcaccagc atcgtaacct ctttgtggta
     1741 aaccatcacc atgaggaggc ttcaacataa attcgttacc gtccttgtcc ttcaatttgt
     1801 cggttaatgg gttgaatctc aagtcaccag caatagcgaa agcagtgact aattctggag
     1861 aagcaacaaa ggcgtgggtt tgtgggttac catcgtttct agaagtgaag tttctgttgt
     1921 aagaggaaac gatggtgttc ttgtcaccct tcttgatatc tcttctgtcc cattgaccaa
     1981 tacatggacc acaagcgtta gccaagacaa taccaccaaa ttccttgaaa gtttccaatt
     2041 gaccgtcacg ttcgatggtg gctctgattt gttcagaacc tggggtaaca gtgaagatgg
     2101 tcttagattt caaaccgtga gcagcagcat ccttgacaat ggaagcagat ctggacatat
     2161 cttcgtaaga ggagttggta caagaaccaa tcaaaccaac tctaacatct aatggccagt
     2221 tgttagcaac ggcaacttcc ttcatcttag aaactggggt agccaaatct ggggtgaatg
     2281 gaccgttgat gtatggttcc aaagtgttca agtcaatttc gacaacttca tcgtattcag
     2341 cgtccttgtc ggcagacaac aagtccttgt ggtataactt agcgaaatca gcaatcttac
     2401 ctctaccggt agcttccaag tattcaatca tggatttgtt gaatgggaaa acagaagtgg
     2461 tggcaccaat ttcagcaccc atgttacaga tggtacccat accggtagca gagaaagtgt
     2521 cgacaccgtc accaaagtat tcgacaatct taccagtacc acccttaaca gtagtgatac
     2581 cagccaattt caagatgatg tcctttggag aagtccaacc gttcatctta ccggtcaact
     2641 taacacccaa aatctttgga gccttcaatt cccatggtct accagccatg acgtcaacag
     2701 catcagcacc accaacaccg atggccaatt gacctagacc accggcattt ggagtgtggg
     2761 agtcagtacc gatgatcaaa gcacctggga aggcatagtt ttccaaaacg atttggtgga
     2821 tgataccgga acctggcttc cagaaaccca tgttgtattt agcagtggca gaagccaaga
     2881 agtcgtagac ttccttgttc aagtcaatgg ctctcttcaa gtccttttca ccaccgactt
     2941 gagcttggat caaatggtca cagtggacgg tgactggctt ggcaacttga ggcaaaccag
     3001 cggacatgaa ttgtaagata gccatttgag cagtagcatc ttgacaagca acacgatctg
     3061 gtcttagttt caagtaggag acacctcttt ggatatcttg accgtgtggg tcatccaagt
     3121 gaccgtacaa aatcttttca gcgtaggtga atggtctgtt caatctcttt ctgacgatat
     3181 ccaaagtttc gacattttgc ttgtagttga tgaaagaatg atcttccaac aagttttggt
     3241 taaccttgga gtctctggtc aagttggaaa cagtcatttt gaatatgtat tacttggtta
     3301 tggttatata tgacaaaaga aaaagaagaa cagaagaata acgcaaggaa gaacaataac
     3361 tgaaattgat agagaagtat tatgtctttg tctttttata ataaatcaag tgcagaaatc
     3421 cgttagacaa catgagggat aaaatttaac gtgggcgaag aagaaggaaa aaagtttttg
     3481 tgagggcgta attgaagcga tctgttgatt gtagattttt tttttttgag gagtcaaagt
     3541 cagaagagaa cagacaaatg gtattaacca tccaatactt ttttggagca acgctaagct
     3601 catgcttttc cattggttac gtgctcagtt gttagatatg gaaagagagg atgctcacgg
     3661 cagcgtgact ccaattgagc ccgaaagaga ggatgccacg ttttcccgac ggctgctaga
     3721 atggaaaaag gaaaaataga agaatcccat tcctatcatt atttacgtaa tgacccacac
     3781 atttttgaga ttttcaacta ttacgtatta cgataatcct gctgtcatta tcattattat
     3841 ctatatcgac gtatgcaacg tatgtgaagc caagtaggca ccaacaggag gcggatggat
     3901 atactgtggt ctggaagatg ccggaaagcg tccccacttc agaagttcct atactttcta
     3961 gagaatagga acttcactat agagtcgaat aagggcgaca ccccctaatt agcccgggcg
     4021 aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctggcag ttccctactc
     4081 tcgcatgggg agtccccaca ctaccatcgg cgctacggcg tttcacttct gagttcggca
     4141 tggggtcagg tgggaccacc gcgctactgc cgccaggcaa acaaggggtg ttatgagcca
     4201 tattcaggta taaatgggct cgcgataatg ttcagaattg gttaattggt tgtaacactg
     4261 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4321 ccctgataaa tgcttcaata atattgaaaa aggaagaata tgagccatat tcaacgggaa
     4381 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     4441 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     4501 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     4561 gtcagactaa actggctgac ggaatttatg ccacttccga ccatcaagca ttttatccgt
     4621 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc gttccaggta
     4681 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     4741 cggttgcact cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgcctc
     4801 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     4861 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     4921 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     4981 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     5041 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     5101 aaatatggta ttgataatcc tgatatgaat aaattgcaat ttcatttgat gctcgatgag
     5161 tttttctaag cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc
     5221 ccttaacgtg agttacgcgc gcgtcgttcc actgagcgtc agaccccgta gaaaagatca
     5281 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5341 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5401 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5461 cccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5521 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5581 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5641 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5701 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5761 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5821 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5881 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     5941 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     6001 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtga
//
