LOCUS       sequence-254091-        5852 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast pathway position 4. ACO transcription unit with the S. 
            paradoxus TDH3 promoter and A. gossypii TEF1 terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5852)
  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 5852)
  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-Psptdh3-ACO-Tagtef1
FEATURES             Location/Qualifiers
     source          1..5852
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    174..207
                     /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)"
     terminator      3034..3231
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    complement(3333..3366)
                     /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(3836..3853)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3836..3852)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3951..3968)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4122..4710)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4202..4221)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4832..5641)
                     /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(5642..5733)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5800..5819
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggactcc
       61 ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact
      121 gggcctttcg cccgggctaa ttagggggtg tcgcccttat tcgactctat agtgaagttc
      181 ctattctcta gaaagtatag gaacttctga agtggggacg ctttccggca tcttccagac
      241 cacagtatat ccatccgcct cctgttggtg cgttttattt ctgctgccat ccgtaaatgc
      301 caggatttga gcgggttaca caatatatct catattttcg gtgtctgggt cattacttta
      361 ctcttggcat ccactaaata tattggatcc tgctttttaa actggcttcc agaaaaaaat
      421 caatggagtg atgcaaactg cctggagtaa aagatgacac aaggcgattg acctacgcat
      481 gtatctatct cattttctta caccttctat ttcattctaa ctctttgatt tggaaaacac
      541 ctaagaaaaa aaaggttgaa atcagttccc tgaaattgtc cccctacttg actaataaat
      601 atataaagac ggtaggtatt gactgtaatt cgtaaatcta tacttcttaa acttcttcaa
      661 atttactttt ttggatagtc ttatttttgg tttcaatacc ccaagaactt agtttcaaat
      721 aaatacacat acaaacaaaa tgactgtttc caacttgacc agagactcca aggttaacca
      781 aaacttgttg gaagatcatt ctttcatcaa ctacaagcaa aatgtcgaaa ctttggatat
      841 cgtcagaaag agattgaaca gaccattcac ctacgctgaa aagattttgt acggtcactt
      901 ggatgaccca cacggtcaag atatccaaag aggtgtctcc tacttgaaac taagaccaga
      961 tcgtgttgct tgtcaagatg ctactgctca aatggctatc ttacaattca tgtccgctgg
     1021 tttgcctcaa gttgccaagc cagtcaccgt ccactgtgac catttgatcc aagctcaagt
     1081 cggtggtgaa aaggacttga agagagccat tgacttgaac aaggaagtct acgacttctt
     1141 ggcttctgcc actgctaaat acaacatggg tttctggaag ccaggttccg gtatcatcca
     1201 ccaaatcgtt ttggaaaact atgccttccc aggtgctttg atcatcggta ctgactccca
     1261 cactccaaat gccggtggtc taggtcaatt ggccatcggt gttggtggtg ctgatgctgt
     1321 tgacgtcatg gctggtagac catgggaatt gaaggctcca aagattttgg gtgttaagtt
     1381 gaccggtaag atgaacggtt ggacttctcc aaaggacatc atcttgaaat tggctggtat
     1441 cactactgtt aagggtggta ctggtaagat tgtcgaatac tttggtgacg gtgtcgacac
     1501 tttctctgct accggtatgg gtaccatctg taacatgggt gctgaaattg gtgccaccac
     1561 ttctgttttc ccattcaaca aatccatgat tgaatacttg gaagctaccg gtagaggtaa
     1621 gattgctgat ttcgctaagt tataccacaa ggacttgttg tctgccgaca aggacgctga
     1681 atacgatgaa gttgtcgaaa ttgacttgaa cactttggaa ccatacatca acggtccatt
     1741 caccccagat ttggctaccc cagtttctaa gatgaaggaa gttgccgttg ctaacaactg
     1801 gccattagat gttagagttg gtttgattgg ttcttgtacc aactcctctt acgaagatat
     1861 gtccagatct gcttccattg tcaaggatgc tgctgctcac ggtttgaaat ctaagaccat
     1921 cttcactgtt accccaggtt ctgaacaaat cagagccacc atcgaacgtg acggtcaatt
     1981 ggaaactttc aaggaatttg gtggtattgt cttggctaac gcttgtggtc catgtattgg
     2041 tcaatgggac agaagagata tcaagaaggg tgacaagaac accatcgttt cctcttacaa
     2101 cagaaacttc acttctagaa acgatggtaa cccacaaacc cacgcctttg ttgcttctcc
     2161 agaattagtc actgctttcg ctattgctgg tgacttgaga ttcaacccat taaccgacaa
     2221 attgaaggac aaggacggta acgaatttat gttgaagcct cctcatggtg atggtttacc
     2281 acaaagaggt tacgatgctg gtgaaaacac ctaccaagct ccaccagccg acagatccac
     2341 cgtcgaagtc aaggtttctc caacttctga cagattacaa ttgttgaaac ctttcaagcc
     2401 atgggatggt aaggacgcta aggacatgcc aatcttaatc aaggctgttg gtaagactac
     2461 caccgaccac atttccatgg ctggtccatg gttgaaatac agaggtcact tggaaaacat
     2521 ctccaacaac tacatgattg gtgccattaa tgccgaaaac aagaaggcta actgtgtcaa
     2581 gaacgtttac actggtgaat acaagggtgt tccagacact gccagagact acagagatca
     2641 aggtatcaaa tgggttgtca tcggtgacga aaacttcggt gaaggttctt ctcgtgaaca
     2701 cgctgctttg gaaccaagat tcttgggtgg tttcgctatt attaccaaat ctttcgctcg
     2761 tattcacgaa accaacttga agaagcaagg tctattgcca ttgaacttca agaacccagc
     2821 cgactacgac aagatcaacc cagatgacag aattgacatc ttaggtttgg ctgaattggc
     2881 tccaggtaag ccagtcacca tgagagttca cccaaagaac ggtaagccat gggatgctgt
     2941 cttgactcac actttcaacg atgaacaaat cgaatggttc aaatacggtt ctgctttgaa
     3001 caagatcaag gctgatgaaa agaagtaaac agtactgaca ataaaaagat tcttgttttc
     3061 aagaacttgt catttgtata gtttttttat attgtagttg ttctatttta atcaaatgtt
     3121 agcgtgattt atattttttt tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg
     3181 cagaaagtaa tatcatgcgt caatcgtatg tgaatgctgg tcgctatact gctgtcgatt
     3241 cgatactaac gccgccatcc agtgtcgacc tcaacgttgt ccaggtttgt atccacgtgt
     3301 gtccgttccg ccaatattcc gcaaaatgaa gtgaagttcc tatactttct agagaatagg
     3361 aacttctata gtgagtcgaa taagggcgac acaaaattta ttctaaatgc ataataaata
     3421 ctgataacat cttatagttt gtattatatt ttgtattatc gttgacatgt ataattttga
     3481 tatcaaaaac tgattttccc tttattattt tcgagattta ttttcttaat tctctttaac
     3541 aaactagaaa tattgtatat acaaaaaatc ataaataata gatgaatagt ttaattatag
     3601 gtgttcatca atcgaaaaag caacgtatct tatttaaagt gcgttgcttt tttctcattt
     3661 ataaggttaa ataattctca tatatcaagc aaagtgacag gcgcccttaa atattctgac
     3721 aaatgctctt tccctaaact ccccccataa aaaaacccgc cgaagcgggt ttttacgtta
     3781 tttgcggatt aacgattact cgttatcaga accgcccagg gggcccgagc ttaagactgg
     3841 ccgtcgtttt acaacacaga aagagtttgt agaaacgcaa aaaggccatc cgtcaggggc
     3901 cttctgctta gtttgatgcc tggcagttcc ctactctcgc cttccgcttc ctcgctcact
     3961 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4021 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4081 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4141 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4201 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4261 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4321 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4381 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4441 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4501 aggtatgtag gcggtgctac agagttcttg aagtggtggg ctaactacgg ctacactaga
     4561 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4621 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4681 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4741 gacgctcagt ggaacgacgc gcgcgtaact cacgttaagg gattttggtc atgagcttgc
     4801 gccgtcccgt caagtcagcg taatgctctg cttagaaaaa ctcatcgagc atcaaatgaa
     4861 attgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
     4921 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
     4981 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
     5041 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat
     5101 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
     5161 catcaaccaa accgttattc attcgtgatt gcgcctgagc gaggcgaaat acgcgatcgc
     5221 tgttaaaagg acaattacaa acaggaatcg agtgcaaccg gcgcaggaac actgccagcg
     5281 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaac gctgtttttc
     5341 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     5401 tcggaagtgg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     5461 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     5521 agcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     5581 aatcagcatc catgttggaa tttaatcgcg gcctcgacgt ttcccgttga atatggctca
     5641 tattcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5701 acatatttga atgtatttag aaaaataaac aaataggggt cagtgttaca accaattaac
     5761 caattctgaa cattatcgcg agcccattta tacctgaata tggctcataa caccccttgt
     5821 ttgcctggcg gcagtagcgc ggtggtccca cc
//
