LOCUS Exported 6577 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Integration of ELO3-mTagBFP2 as an additional copy in genome, use auxotrophic marker TRP1(Kluyveromyces lactis). Marker for yeast endoplasmic reticulum (ER).. ACCESSION . VERSION . KEYWORDS ZJOM136 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6577) AUTHORS Zhu J, Zhang ZT, Tang SW, Zhao BS, Li H, Song JZ, Li D, Xie Z TITLE A Validated Set of Fluorescent-Protein-Based Markers for Major Organelles in Yeast (Saccharomyces cerevisiae). JOURNAL MBio. 2019 Sep 3;10(5). pii: mBio.01691-19. doi: 10.1128/mBio.01691-19. PUBMED 31481383 REFERENCE 2 (bases 1 to 6577) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6577 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..19 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 1..18 /label=SP6 /note="SP6 promoter, forward primer" terminator 2870..3057 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind complement(2925..2949) /label=pHybLex-R /note="ADH terminator, reverse primer" protein_bind complement(3109..3142) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind complement(4145..4178) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind complement(4231..4250) /label=T7 /note="T7 promoter, forward primer" promoter complement(4232..4250) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4337..4354) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4508..5096) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4588..4607) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5267..6127) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 5890..5909 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6128..6232) /gene="bla" /label=AmpR promoter primer_bind 6300..6318 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(6356..6378) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 6478..6497 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 atttaggtga cactatagaa cgcggccgcc agctgaagct tcgtacgctg caggtcgtgg 61 gattcctggg ttaattaaca tcgctagcgt aaaaagggtt ttcagatcgt tagcatttca 121 cagaatgctc atgaatcctt gttagatccg gccgatacat cattagagtt tgcactaagg 181 gatactttga aaccaatggc aatctatcgt gtgggaaata tgtttttgct gtgttacaca 241 gaatttgcct tctttgtcaa taatcagggt tggagaaaga aagaatcgca tattattcat 301 tgggagggag agccccaaaa atttgcaatt tggtacccgt acatcttggc attcgacagc 361 aacttcattg agatcagaaa gattgaaact ggcgaattaa ttcgttgcgt ccttgctgat 421 aagattcgac ttctacaaac aagtactcaa gaaatcctgt actgctacga ggattatcgc 481 gggtatgata cagttgcgtc gttagatttc tggggttaag atagcaacgt atatagactt 541 tccttatttt ctttagaata aaaagcataa ctaaagttgt aaaaggagca gaattatgaa 601 taataaacat tgatgacaat agtggaaaac gacataaata ataattttat aatataataa 661 tgataattca ccgtcagagg gatttgtatg caaatgattg atgacgcgag aaaaaaaaac 721 gcgaaatttt tcttcccaaa gcttcgaagt ttttattccg tccacaaagt gaaaaatttt 781 cagaaaattt tccagactgt gaataaacaa aaggttggct tacgcatttg gcttgtcttt 841 tttttttttc acaggttctt agtaaataat tgtataaaat atttatctgg agtcttctgt 901 ttgttgttta gccttgtttg aagttttatt taacttatta ccccagttag gtactgtgta 961 ttttccgcat atctggtgtc tttttatttg gtcattgatt ggtttttgac agctcttcac 1021 tcgcttcaag cttacttcta gtttatttat tcggcttttt tccgtttgtt tacgaaacat 1081 aaacagtcat gaacactacc acatctactg ttatagcagc agttgccgac cagttccagt 1141 ctttgaactc ttcttcttca tgtttcttga aggttcatgt tccttccatt gagaacccat 1201 tcggtattga attatggcca attttctcca aagtgtttga atactttagt ggctatccag 1261 ctgagcaatt cgagtttatt cacaataaga ctttcttggc taacgggtat catgctgtta 1321 gtattattat cgtttattac attattatct ttggtggcca agctatctta cgcgccttga 1381 acgcctctcc attaaagttt aaattgcttt tcgagataca caacttgttt ttgacttcta 1441 tttctctagt tttatggttg ctgatgttag aacagttggt tcctatggtt tatcacaacg 1501 gtctattctg gtctatctgc tctaaggaag ccttcgcacc aaaattagtt actctttact 1561 atttgaacta tttgaccaaa ttcgtagaat tgattgacac tgtgttttta gttttgagaa 1621 gaaagaagtt attgtttttg cacacttacc atcacggtgc caccgctttg ttgtgctaca 1681 ctcaattaat tggtcgtact tctgttgaat gggtagttat cctactaaac ttgggtgttc 1741 acgttatcat gtactggtac tacttcttga gttcatgtgg tattagagtt tggtggaagc 1801 aatgggtcac tagattccaa attattcaat ttttgattga cttggtattt gtttactttg 1861 ctacctatac attctatgct cacaaatact tggacggtat tttaccaaac aagggtactt 1921 gttatggtac tcaggctgct gctgcttatg ggtatttgat tctaacatct tatttgcttt 1981 tgtttatttc cttctacatc caatcttaca agaaaggtgg taaaaagaca gtcaagaagg 2041 aatctgaagt ttccggctcc gttgcatccg gttcttctac tggtgtcaag acctctaaca 2101 ccaaggtctc ttccaggaaa gctctttctc ttgaagttgc tgaagaaatg agcgaactga 2161 tcaaagagaa catgcacatg aagctgtaca tggaaggcac cgttgacaac caccacttta 2221 agtgcacctc tgagggtgag ggtaagccct acgaaggcac ccaaaccatg cgtatcaaag 2281 ttgtggaggg cggtccactg ccattcgctt ttgacattct ggccaccagc ttcctgtacg 2341 gttccaaaac cttcattaac catactcagg gcattcctga tttcttcaaa cagagctttc 2401 cggaaggttt cacctgggag cgtgtcacca cgtatgaaga tggtggtgtg ttgaccgcca 2461 cccaagatac ctccctgcaa gatggctgtc tgatctataa cgtgaaaatt cgtggcgtca 2521 actttactag caatggtccc gtgatgcaga agaaaaccct gggttgggag gcctttacag 2581 aaaccctgta tcctgccgat ggtggcctgg agggccgtaa cgacatggca ctgaagctgg 2641 ttggtggcag ccatttgatc gcaaatgcca agaccactta cagaagcaag aaacccgcaa 2701 aaaatctgaa gatgccgggt gtttactatg tcgactaccg tctggaaaga attaaagaag 2761 caaataatga gacttacgtg gagcagcacg aggttgcagt cgcgagatat tgcgacttgc 2821 ctagcaagct gggtcataaa ctgaattgag gcgcgccact tctaaataag cgaatttctt 2881 atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat acaaatttta 2941 aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt tcctgtaggt 3001 caggttgctt tctcaggtat agtatgaggt cgctcttatt gaccacacct ctaccggcag 3061 atccgctagg gataacaggg taatatctgc aggtcgacaa cccttaatat aacttcgtat 3121 aatgtatgct atacgaagtt attaggtcta gagcggccgc ggaacgatca ttcactatat 3181 atatatcaat ttatatatac gtatgtgtaa ttgaagaaag atacgttttt tcctctattg 3241 agaggcctgc tggatgaata gctttacctt ttctaaatcc ttgataccat cagtctctac 3301 tcctccactt acatcgacac cgatcgcatt tggtaacata ttaatggcaa cagaaacgtt 3361 atcaggattc aatccaccag cgataatgaa ttttatctcg ggatgacttg cagaccaact 3421 ggaaattgca ctccaattca atttctcacc agtgccacct tcaccagaat cgaacaacgt 3481 cagcacattg tctacgtgtt catacaggtc cagtagtaat tcacaatcct gtgggaactg 3541 gaacctctta atgattggaa ttgaagatgg gatcaaagat ctgtattctt taatatcttc 3601 atctccatgt aattgtatca catctagatt atattcgtgg tacagttgaa ggacatcatc 3661 aacggactga tttctaaaca ccccgactag tttagtacct ttcacgttct cttgttggtg 3721 aactgcagtt gaaatacctt tcgcaacaga tgagtcaatg gttcttttcc taccgggaac 3781 acaaatgata cctaagtaat cagcaccatc atccacagca gtctttgcag cttcaacggt 3841 ttgcaaacca cacactttaa cgagcatcac agtagatttc gcaagagggt agctcgctca 3901 gagtacccaa gtaaatgatt agtaaactga tgtttgatag ttcaattttt caatgaaata 3961 accttatatt aaaattgata ttactattat acaaaaataa agaataaagg atttgagttt 4021 atacataaaa taccattatt atttgttcag tgagagatac tggggtatat gggatgtgtg 4081 tagtgatacc atgcaatcat gtatcaaaca tgggcccggt accgagctct cgagaaccct 4141 taatataact tcgtataatg tatgctatac gaagttatta ggtgatatca gatccactag 4201 tggcctatgc ggccgcggat ctgccggtct ccctatagtg agtcgtatta atttcgataa 4261 gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 4321 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 4381 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 4441 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 4501 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 4561 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 4621 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 4681 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 4741 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 4801 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 4861 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 4921 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc 4981 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 5041 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 5101 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 5161 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 5221 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 5281 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 5341 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 5401 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 5461 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 5521 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 5581 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 5641 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 5701 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 5761 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 5821 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 5881 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 5941 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 6001 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 6061 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 6121 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 6181 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 6241 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa 6301 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct 6361 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac 6421 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg 6481 catcagagca gattgtactg agagtgcacc atatggacat attgtcgtta gaacgcggct 6541 acaattaata cataacctta tgtatcatac acatacg //