LOCUS       sequence-254145-        5165 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast pathway position 7. ACDH transcription unit with the ENO2 
            promoter and TAL1 terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5165)
  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 5165)
  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-Peno2-ACDH-Ttal1
FEATURES             Location/Qualifiers
     source          1..5165
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    126..159
                     /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(2598..2631)
                     /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(3101..3118)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3101..3117)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3216..3233)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3387..3975)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3467..3486)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4097..4906)
                     /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(4907..4998)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5065..5084
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tgtggggact ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc
       61 agtcgaaaga ctgggccttt cgcccgggct aattaggggg tgtcgccctt attcgactct
      121 atagtgaagt tcctattctc tagaaagtat aggaacttct gaagtgggga aaccttcgaa
      181 tccagccagc atgtcgacac ccacaagatg tagtgcacgt gcgtgtcgac gctgcgggta
      241 tagaaagggt tctttactct atagtacctc ctcgctcagc atctgcttct tcccaaagat
      301 gaacgcggcg ttatgtcact aacgacgtgc accaacttgc ggaaagtgga atcccgttcc
      361 aaaactggca tccactaatt gatacatcta cacaccgcac gccttttttc tgaagcccac
      421 tttcgtggac tttgccatat gcaaaattca tgaagtgtga taccaagtca gcatacacct
      481 cactagggta gtttctttgg ttgtattgat catttggttc atcgtggttc attaattttt
      541 tttctccatt gctttctggc tttgatctta ctatcatttg gatttttgtc gaaggttgta
      601 gaattgtatg tgacaagtgg caccaagcat atataaaaaa aaaaagcatt atcttcctac
      661 cagagttgat tgttaaaaac gtatttatag caaacgcaat tgtaattaat tcttattttg
      721 tatcttttct tcccttgtct caatctttta tttttatttt atttttcttt tcttagtttc
      781 tttcataaca ccaagcaact aatactataa catacaataa taatggaatc tttggaattg
      841 gaacaattag tcaagaaggt tttgttggaa aaattggctg aacaaaagga agttccaacc
      901 aagaccacca cccaaggtgc caagtccggt gttttcgaca ccgtcgatga agctgtccaa
      961 gctgctgtca ttgctcaaaa ctgttacaag gaaaaatctt tggaagaaag aagaaacgtt
     1021 gtcaaggcca tcagagaagc tttgtaccca gaaatcgaaa ccattgccac cagagctgtt
     1081 gctgaaaccg gtatgggtaa tgtcactgac aagatcttga agaacacttt ggccatcgaa
     1141 aagaccccag gtgttgaaga tttgtacact gaagttgcca ctggtgacaa cggtatgact
     1201 ttgtacgaat tgtctccata cggtgtcatc ggtgctgttg ccccatctac caacccaact
     1261 gaaactttga tctgtaactc cattggtatg ttggctgctg gtaatgctgt tttctactct
     1321 cctcacccag gtgccaagaa catctcttta tggttaatcg aaaaattgaa caccattgtc
     1381 cgtgactctt gtggtatcga caacttgatt gtcactgttg ccaagccttc catccaagct
     1441 gctcaagaaa tgatgaacca tccaaaggtc ccattgttgg ttatcactgg tggtccaggt
     1501 gttgtcttgc aagctatgca atctggtaag aaggtcattg gtgctggtgc tggtaaccca
     1561 ccatctatcg tcgatgaaac tgctaacatt gaaaaggctg ccgctgatat cgttgacggt
     1621 gcttctttcg accacaacat cctatgtatt gctgaaaaat ccgttgttgc cgttgactcc
     1681 attgctgatt tcttattatt ccaaatggaa aagaacggtg ctttgcacgt taccaaccca
     1741 tctgatatcc aaaaattgga aaaggttgct gtcactgaca agggtgtcac caacaagaaa
     1801 ttggttggta agtctgctac tgaaatcttg aaggaagctg gtattgcttg tgacttcact
     1861 ccaagattaa tcattgtcga aactgaaaag tcccacccat ttgccaccgt tgaattgttg
     1921 atgccaattg tcccagttgt cagagttcca gacttcgatg aagctttgga agttgccatc
     1981 gaattggaac aaggtttgca ccacactgct accatgcact ctcaaaacat ctccagattg
     2041 aacaaggctg ctagagacat gcaaacttcc atctttgtca agaacggtcc atctttcgct
     2101 ggtttaggtt tcagaggtga aggttccact actttcacca ttgctacccc aactggtgaa
     2161 ggtaccacca ccgctagaca tttcgctaga agaagaagat gtgttttgac tgatggtttc
     2221 tccatacgtt aaaggaagta tctcggaaat attaatttag gccatgtcct tatgcacgtt
     2281 tcttttgata cttacgggta catgtacaca agtatatcta tatatataaa ttaatgaaaa
     2341 tcccctattt atatatatga ctttaacgag acagaacagt tttttatttt ttatcctatt
     2401 tgatgaatga tacagtttct tattcacgtg ttatacccac accaaatcca atagcaatac
     2461 cggccatcac aatcactgtt tcggcagccc ctaagatcag acaaaacatc cggaaccacc
     2521 ttaaatcaac gtccctcaga aagcctgtat gcgaagccac aatcctttcc aacagaccat
     2581 actaagtaaa atgaagtgaa gttcctatac tttctagaga ataggaactt ctatagtgag
     2641 tcgaataagg gcgacacaaa atttattcta aatgcataat aaatactgat aacatcttat
     2701 agtttgtatt atattttgta ttatcgttga catgtataat tttgatatca aaaactgatt
     2761 ttccctttat tattttcgag atttattttc ttaattctct ttaacaaact agaaatattg
     2821 tatatacaaa aaatcataaa taatagatga atagtttaat tataggtgtt catcaatcga
     2881 aaaagcaacg tatcttattt aaagtgcgtt gcttttttct catttataag gttaaataat
     2941 tctcatatat caagcaaagt gacaggcgcc cttaaatatt ctgacaaatg ctctttccct
     3001 aaactccccc cataaaaaaa cccgccgaag cgggttttta cgttatttgc ggattaacga
     3061 ttactcgtta tcagaaccgc ccagggggcc cgagcttaag actggccgtc gttttacaac
     3121 acagaaagag tttgtagaaa cgcaaaaagg ccatccgtca ggggccttct gcttagtttg
     3181 atgcctggca gttccctact ctcgccttcc gcttcctcgc tcactgactc gctgcgctcg
     3241 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3301 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3361 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3421 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3481 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3541 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3601 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3661 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3721 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3781 gctacagagt tcttgaagtg gtgggctaac tacggctaca ctagaagaac agtatttggt
     3841 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3901 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3961 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4021 gacgcgcgcg taactcacgt taagggattt tggtcatgag cttgcgccgt cccgtcaagt
     4081 cagcgtaatg ctctgcttag aaaaactcat cgagcatcaa atgaaattgc aatttattca
     4141 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     4201 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     4261 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     4321 caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga
     4381 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     4441 tattcattcg tgattgcgcc tgagcgaggc gaaatacgcg atcgctgtta aaaggacaat
     4501 tacaaacagg aatcgagtgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     4561 cacctgaatc aggatattct tctaatacct ggaacgctgt ttttccgggg atcgcagtgg
     4621 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agtggcataa
     4681 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     4741 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaagcga tagattgtcg
     4801 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     4861 tggaatttaa tcgcggcctc gacgtttccc gttgaatatg gctcatattc ttcctttttc
     4921 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4981 tttagaaaaa taaacaaata ggggtcagtg ttacaaccaa ttaaccaatt ctgaacatta
     5041 tcgcgagccc atttatacct gaatatggct cataacaccc cttgtttgcc tggcggcagt
     5101 agcgcggtgg tcccacctga ccccatgccg aactcagaag tgaaacgccg tagcgccgat
     5161 ggtag
//
