LOCUS       sequence-254149-        5027 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast pathway position 7. ACDH transcription unit with the RPL28 
            promoter and ECM10 terminator..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5027)
  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 5027)
  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-Prpl28-ACDH-Tecm10
FEATURES             Location/Qualifiers
     source          1..5027
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    187..220
                     /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(2521..2554)
                     /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(3024..3041)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3024..3040)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3139..3156)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3310..3898)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3390..3409)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4020..4829)
                     /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(4830..4921)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4988..5007
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta
       61 gtgtggggac tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcgaaag actgggcctt tcgcccgggc taattagggg gtgtcgccct tattcgactc
      181 tatagtgaag ttcctattct ctagaaagta taggaacttc tgaagtgggg aaaccttcga
      241 atccagccag catgtcgaca cccacaagat gtagtgcacg tgcttaatca tcgtttactg
      301 ccgcctatga gcgtaagcta atgttataaa gaaacaagct ataatattgt taaatatagt
      361 tgatcaacag cattgtaatg attacaagag acgaggtgga atgaacctta tgaaatgcgt
      421 tttatatata aactgtaata agagctaagt tgaattgaaa tctacgatac ttgatgttga
      481 cattatagca ctagttccca ggaaaccctt tcgaaaaaca cagcaaaaac aagagtactg
      541 taaccaatgt aacatctgta caccagggac ccacacatta ccaaaatcaa aattattttt
      601 ctaatgcctg ttatttttcc tatttttcct ctggcgcgtg aatagcccgc agagacgcaa
      661 acaattttcc tcgcagtttt tcgcttgttt aatgcgtatt ttcccagata ggttcaaacc
      721 tttcatctgt atcccgtata tttaagatgg cgtttgcttt ctccgttgat ttttttcctt
      781 cttagtgatt tttttgcatt aaatcccaga acaatcatcc aactaatcaa gaatggaatc
      841 tttggaattg gaacaattag tcaagaaggt tttgttggaa aaattggctg aacaaaagga
      901 agttccaacc aagaccacca cccaaggtgc caagtccggt gttttcgaca ccgtcgatga
      961 agctgtccaa gctgctgtca ttgctcaaaa ctgttacaag gaaaaatctt tggaagaaag
     1021 aagaaacgtt gtcaaggcca tcagagaagc tttgtaccca gaaatcgaaa ccattgccac
     1081 cagagctgtt gctgaaaccg gtatgggtaa tgtcactgac aagatcttga agaacacttt
     1141 ggccatcgaa aagaccccag gtgttgaaga tttgtacact gaagttgcca ctggtgacaa
     1201 cggtatgact ttgtacgaat tgtctccata cggtgtcatc ggtgctgttg ccccatctac
     1261 caacccaact gaaactttga tctgtaactc cattggtatg ttggctgctg gtaatgctgt
     1321 tttctactct cctcacccag gtgccaagaa catctcttta tggttaatcg aaaaattgaa
     1381 caccattgtc cgtgactctt gtggtatcga caacttgatt gtcactgttg ccaagccttc
     1441 catccaagct gctcaagaaa tgatgaacca tccaaaggtc ccattgttgg ttatcactgg
     1501 tggtccaggt gttgtcttgc aagctatgca atctggtaag aaggtcattg gtgctggtgc
     1561 tggtaaccca ccatctatcg tcgatgaaac tgctaacatt gaaaaggctg ccgctgatat
     1621 cgttgacggt gcttctttcg accacaacat cctatgtatt gctgaaaaat ccgttgttgc
     1681 cgttgactcc attgctgatt tcttattatt ccaaatggaa aagaacggtg ctttgcacgt
     1741 taccaaccca tctgatatcc aaaaattgga aaaggttgct gtcactgaca agggtgtcac
     1801 caacaagaaa ttggttggta agtctgctac tgaaatcttg aaggaagctg gtattgcttg
     1861 tgacttcact ccaagattaa tcattgtcga aactgaaaag tcccacccat ttgccaccgt
     1921 tgaattgttg atgccaattg tcccagttgt cagagttcca gacttcgatg aagctttgga
     1981 agttgccatc gaattggaac aaggtttgca ccacactgct accatgcact ctcaaaacat
     2041 ctccagattg aacaaggctg ctagagacat gcaaacttcc atctttgtca agaacggtcc
     2101 atctttcgct ggtttaggtt tcagaggtga aggttccact actttcacca ttgctacccc
     2161 aactggtgaa ggtaccacca ccgctagaca tttcgctaga agaagaagat gtgttttgac
     2221 tgatggtttc tccatacgtt aaaacaagat aataaaaaag ataatatttt cgtttaaaat
     2281 ttcagaaata ttgcttacat caaacgaaat agtaagcgta aaccatatat cctttcaacg
     2341 atatgtatca tttttatagt cttttggcgg tataaaaaga atagccaaag gtggaagtca
     2401 ggttaaacca aaagaaacta cccgcaaggg atgtatgcat ttgaacataa aaaactcctc
     2461 agaaagcctg tatgcgaagc cacaatcctt tccaacagac catactaagt aaaatgaagt
     2521 gaagttccta tactttctag agaataggaa cttctatagt gagtcgaata agggcgacac
     2581 aaaatttatt ctaaatgcat aataaatact gataacatct tatagtttgt attatatttt
     2641 gtattatcgt tgacatgtat aattttgata tcaaaaactg attttccctt tattattttc
     2701 gagatttatt ttcttaattc tctttaacaa actagaaata ttgtatatac aaaaaatcat
     2761 aaataataga tgaatagttt aattataggt gttcatcaat cgaaaaagca acgtatctta
     2821 tttaaagtgc gttgcttttt tctcatttat aaggttaaat aattctcata tatcaagcaa
     2881 agtgacaggc gcccttaaat attctgacaa atgctctttc cctaaactcc ccccataaaa
     2941 aaacccgccg aagcgggttt ttacgttatt tgcggattaa cgattactcg ttatcagaac
     3001 cgcccagggg gcccgagctt aagactggcc gtcgttttac aacacagaaa gagtttgtag
     3061 aaacgcaaaa aggccatccg tcaggggcct tctgcttagt ttgatgcctg gcagttccct
     3121 actctcgcct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3181 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3241 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3301 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3361 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3421 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3481 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3541 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3601 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3661 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3721 gtggtgggct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3781 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3841 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3901 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgacgcgc gcgtaactca
     3961 cgttaaggga ttttggtcat gagcttgcgc cgtcccgtca agtcagcgta atgctctgct
     4021 tagaaaaact catcgagcat caaatgaaat tgcaatttat tcatatcagg attatcaata
     4081 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
     4141 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
     4201 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
     4261 gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag
     4321 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc
     4381 gcctgagcga ggcgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgag
     4441 tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat
     4501 tcttctaata cctggaacgc tgtttttccg gggatcgcag tggtgagtaa ccatgcatca
     4561 tcaggagtac ggataaaatg cttgatggtc ggaagtggca taaattccgt cagccagttt
     4621 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac
     4681 aactctggcg catcgggctt cccatacaag cgatagattg tcgcacctga ttgcccgaca
     4741 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc
     4801 ctcgacgttt cccgttgaat atggctcata ttcttccttt ttcaatatta ttgaagcatt
     4861 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4921 ataggggtca gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata
     4981 cctgaatatg gctcataaca ccccttgttt gcctggcggc agtagcg
//
