LOCUS Exported 7049 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Yeast pathway position 5. PYC transcription unit with the HHF2 promoter and ADH1 terminator.. ACCESSION . VERSION . KEYWORDS pDSM-de-Phhf2-PYC-Tadh1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7049) 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 7049) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7049 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind complement(3965..3998) /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(4468..4485) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4468..4484) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4583..4600) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4754..5342) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4834..4853) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5464..6273) /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(6274..6365) /gene="bla" /label=AmpR promoter primer_bind 6432..6451 /label=pENTR-R /note="pENTR vectors, reverse primer" protein_bind 6658..6691 /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)" ORIGIN 1 atgcagacca gaccagaaca aaacaactga caagaaggcg tttaatttaa tatgttgttc 61 actcgcgcct gggctgttgt tattcggcta gatacatacg tgtttgtgcg tatgtagtta 121 tatcatatat aagtatatta ggatgaggcg gtgaaagaga tttttttttt ttcgcttaat 181 ttattctttt ctctatcttt tttcctacat cttgttcaaa agagtagcaa aaacaacaat 241 caatacaata aaataatgtc ctcttccaag atcttggctg gtttgagaga caacttttct 301 ttgttgggtg aaaagaacaa gattttggtc gccaacagag gtgaaatccc aatcagaatt 361 ttcagatctg ctcacgaatt gtctatgaga actatcgcca tctactctca cgaagataga 421 ttatccatgc acagattgaa ggctgatgaa gcctacgtta tcggtgaaga aggtcaatac 481 accccagtcg gtgcttactt ggccatggac gaaatcatcg aaattgccaa gaagcacaag 541 gtcgatttca tccacccagg ttacggtttc ttgtctgaaa actctgaatt tgctgacaag 601 gttgttaagg ctggtattac ctggattggt ccaccagctg aagtcattga atctgttggt 661 gacaaggttt ctgccagaca tttggctgct cgtgccaacg ttccaactgt cccaggtact 721 ccaggtccta tcgaaaccgt tcaagaagct ctagatttcg tcaatgaata cggttaccca 781 gttatcatca aggctgcttt cggtggtggt ggtcgtggta tgagagttgt cagagaaggt 841 gacgatgtcg ctgatgcttt ccaaagagcc acttctgaag ctagaactgc tttcggtaac 901 ggtacttgtt tcgtcgaaag attcttggac aagccaaagc acattgaagt tcaattatta 961 gctgacaacc acggtaacgt tgtccacttg ttcgaaagag actgttccgt ccaaagacgt 1021 caccaaaagg ttgtcgaagt tgctccagct aagactttac caagagaagt tagagatgct 1081 atcttgaccg atgccgttaa gttggctaag gtttgtggtt acagaaacgc tggtactgct 1141 gaattcttgg ttgacaacca aaacagacat tacttcattg aaatcaaccc aagaattcaa 1201 gtcgaacaca ccatcactga agaaatcact ggtattgaca ttgtctccgc tcaaatccaa 1261 atcgccgctg gtgctacttt gactcaatta ggtctattac aagacaaaat caccaccaga 1321 ggtttctcta tccaatgtcg tatcaccact gaagatccat ccaagaactt ccaaccagac 1381 actggtcgtt tggaagtcta cagatccgct ggtggtaacg gtgtcagatt ggacggtggt 1441 aacgcctacg ctggtgctac catctctcca cactacgact ccatgttggt taagtgttcc 1501 tgttctggtt ctacctacga aattgtcaga agaaagatga tcagagcttt gattgaattc 1561 agaatcagag gtgtcaagac caacatccca ttcttgttga ctttgttgac caacccagtt 1621 ttcattgaag gtacctactg gaccactttc atcgatgaca ctccacaatt gttccaaatg 1681 gtttcctctc aaaacagagc tcaaaaattg ttgcactact tggctgactt ggccgtcaac 1741 ggttcctcta tcaagggtca aatcggttta ccaaagttga agtccaaccc ttccgttcca 1801 catttgcacg atgctcaagg taatgtcatc aacgttacca aatctgcccc accatccggt 1861 tggagacaag tcttgttgga aaagggtcca tccgaatttg ccaagcaagt cagacaattc 1921 aacggtactt tgttgatgga caccacctgg agagatgctc accaatcttt gctagctacc 1981 agagtcagaa ctcacgattt ggccaccatt gctccaacca ctgctcacgc tttggctggt 2041 gcctttgctt tggaatgttg gggtggtgct actttcgatg tcgccatgag attcttgcat 2101 gaggacccat gggaaagatt gagaaaattg agatctttgg tcccaaacat tccattccaa 2161 atgttgttga gaggtgctaa cggtgttgct tactcctctt tgccagacaa cgccattgac 2221 catttcgtta agcaagccaa ggacaatggt gttgacattt tcagagtctt tgacgctttg 2281 aacgacttgg aacaattgaa ggttggtgtt aatgctgtca agaaggctgg tggtgttgtc 2341 gaagctaccg tttgttactc tggtgacatg ttgcaaccag gtaagaaata caacttggac 2401 tactacttag aagttgtcga aaagatcgtt caaatgggta ctcacatctt gggtatcaag 2461 gacatggctg gtaccatgaa gccagctgct gccaaattgt tgattggttc tttacgtacc 2521 agatacccag acttgccaat ccacgttcac tctcatgact ccgctggtac tgctgttgct 2581 tccatgactg cttgtgcttt ggccggtgct gatgttgttg acgttgccat taactccatg 2641 tccggtttga cctctcaacc atctattaac gctttgttgg cctccttgga aggtaacatt 2701 gacactggta tcaacgtcga acacgttaga gaattggacg cttactgggc tgaaatgaga 2761 ttattatact cttgtttcga agctgacttg aagggtccag accctgaagt ttaccaacac 2821 gaaattccag gtggtcaatt gaccaacttg ttgttccaag ctcaacaatt aggtctaggt 2881 gaacaatggg ctgaaaccaa gagagcttac agagaagcta actacttgtt gggtgacatt 2941 gttaaggtca ccccaacttc taaggtcgtt ggtgatttgg ctcaattcat ggtttctaac 3001 aaattgactt ctgatgacat cagaagatta gctaactctt tggacttccc agactccgtt 3061 atggacttct tcgaaggttt gatcggtcaa ccatacggtg gtttcccaga accattgaga 3121 tccgatgttt tgagaaacaa gcgtcgtaaa ttgacttgta gaccaggttt agaattggaa 3181 ccattcgatt tggaaaagat cagagaagat ttgcaaaaca gattcggtga tatcgatgaa 3241 tgtgatgttg cctcctacaa catgtatcct cgtgtctacg aagatttcca aaagattaga 3301 gaaacttacg gtgacttgtc tgtcttacca accaagaact tcttggctcc agctgaacca 3361 gacgaagaaa tcgaagtcac cattgaacaa ggtaagactt tgattatcaa attacaagct 3421 gttggtgatt tgaacaagaa aaccggtcaa agagaagtct acttcgaatt gaacggtgaa 3481 ttgagaaaga tcagagttgc tgacaaatct caaaacattc aatctgttgc caagccaaag 3541 gctgatgtcc acgacaccca ccaaatcggt gctccaatgg ctggtgtcat cattgaagtc 3601 aaggttcaca agggttcttt ggtcaagaag ggtgaatcta tcgccgtttt gtctgctatg 3661 aagatggaaa tggttgtttc ctctccagct gatggtcaag tcaaagatgt ctttatccgt 3721 gacggtgaat ccgtcgatgc ttctgacttg ttggttgttt tggaagaaga aactctacca 3781 ccttctcaaa agaaataaac gaatttctta tgatttatga tttttattat taaataagtt 3841 ataaaaaaaa taagtgtata caaattttaa agtgacactt aggttttaaa acgaaaattc 3901 cctcaaataa ccacaaacat ccttcccata tgctcggtcg tgcttgttgt acctaaaatg 3961 aagtgaagtt cctatacttt ctagagaata ggaacttcta tagtgagtcg aataagggcg 4021 acacaaaatt tattctaaat gcataataaa tactgataac atcttatagt ttgtattata 4081 ttttgtatta tcgttgacat gtataatttt gatatcaaaa actgattttc cctttattat 4141 tttcgagatt tattttctta attctcttta acaaactaga aatattgtat atacaaaaaa 4201 tcataaataa tagatgaata gtttaattat aggtgttcat caatcgaaaa agcaacgtat 4261 cttatttaaa gtgcgttgct tttttctcat ttataaggtt aaataattct catatatcaa 4321 gcaaagtgac aggcgccctt aaatattctg acaaatgctc tttccctaaa ctccccccat 4381 aaaaaaaccc gccgaagcgg gtttttacgt tatttgcgga ttaacgatta ctcgttatca 4441 gaaccgccca gggggcccga gcttaagact ggccgtcgtt ttacaacaca gaaagagttt 4501 gtagaaacgc aaaaaggcca tccgtcaggg gccttctgct tagtttgatg cctggcagtt 4561 ccctactctc gccttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 4621 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 4681 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 4741 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 4801 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 4861 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 4921 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt 4981 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 5041 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 5101 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 5161 tgaagtggtg ggctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc 5221 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 5281 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 5341 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgac gcgcgcgtaa 5401 ctcacgttaa gggattttgg tcatgagctt gcgccgtccc gtcaagtcag cgtaatgctc 5461 tgcttagaaa aactcatcga gcatcaaatg aaattgcaat ttattcatat caggattatc 5521 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt 5581 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca 5641 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac 5701 gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg 5761 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga 5821 ttgcgcctga gcgaggcgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat 5881 cgagtgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg 5941 atattcttct aatacctgga acgctgtttt tccggggatc gcagtggtga gtaaccatgc 6001 atcatcagga gtacggataa aatgcttgat ggtcggaagt ggcataaatt ccgtcagcca 6061 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag 6121 aaacaactct ggcgcatcgg gcttcccata caagcgatag attgtcgcac ctgattgccc 6181 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg 6241 cggcctcgac gtttcccgtt gaatatggct catattcttc ctttttcaat attattgaag 6301 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 6361 acaaataggg gtcagtgtta caaccaatta accaattctg aacattatcg cgagcccatt 6421 tatacctgaa tatggctcat aacacccctt gtttgcctgg cggcagtagc gcggtggtcc 6481 cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtgggga 6541 ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa 6601 gactgggcct ttcgcccggg ctaattaggg ggtgtcgccc ttattcgact ctatagtgaa 6661 gttcctattc tctagaaagt ataggaactt ctgaagtggg gaacgttgtc caggtttgta 6721 tccacgtgtg tccgttccgc caatattccg cgtgcacaag aagcaacgcg agagagcaca 6781 acacgctgtt atcacgcaaa ctatgttttg acaccgagcc atagccgtga ttgtgcgtca 6841 cattgggcga taatgaacgc taaatgacca actcccatcc gtaggagccc cttagggcgt 6901 gccaatagtt tcacgcgctt aatgcgaagt gctcggaacg gacaactgtg gtcgtttggc 6961 accgggaaag tggtactaga ccgagagttt cgcatttgta tggcaggacg ttctgggagc 7021 ttcgcgtctc aagctttttc gggcgcgaa //