LOCUS Exported 6575 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Integration of ELO3-mCherry as an additional copy in genome, use auxotrophic marker TRP1(Kluyveromyces lactis). Marker for yeast endoplasmic reticulum (ER).. ACCESSION . VERSION . KEYWORDS ZJOM149 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6575) 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 6575) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6575 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 1720..2430 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(1865..1883) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 2133..2152 /label=mCherry-F /note="mCherry, forward primer" terminator 2460..2647 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind complement(2515..2539) /label=pHybLex-R /note="ADH terminator, reverse primer" protein_bind complement(2699..2732) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind complement(3735..3768) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind complement(3821..3840) /label=T7 /note="T7 promoter, forward primer" promoter complement(3822..3840) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3927..3944) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4098..4686) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4178..4197) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4857..5717) /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 5480..5499 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5718..5822) /gene="bla" /label=AmpR promoter primer_bind 5890..5908 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(5946..5968) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 6068..6087 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 6168..6186 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 6168..6185 /label=SP6 /note="SP6 promoter, forward primer" ORIGIN 1 cctgtactgc tacgaggatt atcgcgggta tgatacagtt gcgtcgttag atttctgggg 61 ttaagatagc aacgtatata gactttcctt attttcttta gaataaaaag cataactaaa 121 gttgtaaaag gagcagaatt atgaataata aacattgatg acaatagtgg aaaacgacat 181 aaataataat tttataatat aataatgata attcaccgtc agagggattt gtatgcaaat 241 gattgatgac gcgagaaaaa aaaacgcgaa atttttcttc ccaaagcttc gaagttttta 301 ttccgtccac aaagtgaaaa attttcagaa aattttccag actgtgaata aacaaaaggt 361 tggcttacgc atttggcttg tctttttttt ttttcacagg ttcttagtaa ataattgtat 421 aaaatattta tctggagtct tctgtttgtt gtttagcctt gtttgaagtt ttatttaact 481 tattacccca gttaggtact gtgtattttc cgcatatctg gtgtcttttt atttggtcat 541 tgattggttt ttgacagctc ttcactcgct tcaagcttac ttctagttta tttattcggc 601 ttttttccgt ttgtttacga aacataaaca gtcatgaaca ctaccacatc tactgttata 661 gcagcagttg ccgaccagtt ccagtctttg aactcttctt cttcatgttt cttgaaggtt 721 catgttcctt ccattgagaa cccattcggt attgaattat ggccaatttt ctccaaagtg 781 tttgaatact ttagtggcta tccagctgag caattcgagt ttattcacaa taagactttc 841 ttggctaacg ggtatcatgc tgttagtatt attatcgttt attacattat tatctttggt 901 ggccaagcta tcttacgcgc cttgaacgcc tctccattaa agtttaaatt gcttttcgag 961 atacacaact tgtttttgac ttctatttct ctagttttat ggttgctgat gttagaacag 1021 ttggttccta tggtttatca caacggtcta ttctggtcta tctgctctaa ggaagccttc 1081 gcaccaaaat tagttactct ttactatttg aactatttga ccaaattcgt agaattgatt 1141 gacactgtgt ttttagtttt gagaagaaag aagttattgt ttttgcacac ttaccatcac 1201 ggtgccaccg ctttgttgtg ctacactcaa ttaattggtc gtacttctgt tgaatgggta 1261 gttatcctac taaacttggg tgttcacgtt atcatgtact ggtactactt cttgagttca 1321 tgtggtatta gagtttggtg gaagcaatgg gtcactagat tccaaattat tcaatttttg 1381 attgacttgg tatttgttta ctttgctacc tatacattct atgctcacaa atacttggac 1441 ggtattttac caaacaaggg tacttgttat ggtactcagg ctgctgctgc ttatgggtat 1501 ttgattctaa catcttattt gcttttgttt atttccttct acatccaatc ttacaagaaa 1561 ggtggtaaaa agacagtcaa gaaggaatct gaagtttccg gctccgttgc atccggttct 1621 tctactggtg tcaagacctc taacaccaag gtctcttcca ggaaagctgc ttcgtacgct 1681 gcaggtcgtg ggattcctgg gttaattaac atcgctagca tggtgagcaa gggcgaggag 1741 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg 1801 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag 1861 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc 1921 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac 1981 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc 2041 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg 2101 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc 2161 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag 2221 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag 2281 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc 2341 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc 2401 accggcggca tggacgagct gtacaagtag cctaggtgag gcgcgccact tctaaataag 2461 cgaatttctt atgatttatg atttttatta ttaaataagt tataaaaaaa ataagtgtat 2521 acaaatttta aagtgactct taggttttaa aacgaaaatt cttattcttg agtaactctt 2581 tcctgtaggt caggttgctt tctcaggtat agtatgaggt cgctcttatt gaccacacct 2641 ctaccggcag atccgctagg gataacaggg taatatctgc aggtcgacaa cccttaatat 2701 aacttcgtat aatgtatgct atacgaagtt attaggtcta gagcggccgc ggaacgatca 2761 ttcactatat atatatcaat ttatatatac gtatgtgtaa ttgaagaaag atacgttttt 2821 tcctctattg agaggcctgc tggatgaata gctttacctt ttctaaatcc ttgataccat 2881 cagtctctac tcctccactt acatcgacac cgatcgcatt tggtaacata ttaatggcaa 2941 cagaaacgtt atcaggattc aatccaccag cgataatgaa ttttatctcg ggatgacttg 3001 cagaccaact ggaaattgca ctccaattca atttctcacc agtgccacct tcaccagaat 3061 cgaacaacgt cagcacattg tctacgtgtt catacaggtc cagtagtaat tcacaatcct 3121 gtgggaactg gaacctctta atgattggaa ttgaagatgg gatcaaagat ctgtattctt 3181 taatatcttc atctccatgt aattgtatca catctagatt atattcgtgg tacagttgaa 3241 ggacatcatc aacggactga tttctaaaca ccccgactag tttagtacct ttcacgttct 3301 cttgttggtg aactgcagtt gaaatacctt tcgcaacaga tgagtcaatg gttcttttcc 3361 taccgggaac acaaatgata cctaagtaat cagcaccatc atccacagca gtctttgcag 3421 cttcaacggt ttgcaaacca cacactttaa cgagcatcac agtagatttc gcaagagggt 3481 agctcgctca gagtacccaa gtaaatgatt agtaaactga tgtttgatag ttcaattttt 3541 caatgaaata accttatatt aaaattgata ttactattat acaaaaataa agaataaagg 3601 atttgagttt atacataaaa taccattatt atttgttcag tgagagatac tggggtatat 3661 gggatgtgtg tagtgatacc atgcaatcat gtatcaaaca tgggcccggt accgagctct 3721 cgagaaccct taatataact tcgtataatg tatgctatac gaagttatta ggtgatatca 3781 gatccactag tggcctatgc ggccgcggat ctgccggtct ccctatagtg agtcgtatta 3841 atttcgataa gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg gagaggcggt 3901 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg 3961 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 4021 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 4081 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 4141 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 4201 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 4261 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 4321 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 4381 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 4441 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 4501 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct 4561 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 4621 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 4681 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 4741 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat 4801 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac 4861 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt 4921 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt 4981 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag 5041 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct 5101 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt 5161 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc 5221 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt 5281 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg 5341 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg 5401 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct 5461 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc 5521 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt 5581 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt 5641 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg 5701 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat 5761 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg 5821 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta 5881 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt 5941 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc 6001 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt 6061 aactatgcgg catcagagca gattgtactg agagtgcacc atatggacat attgtcgtta 6121 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac 6181 tatagaacgc ggccgccagc tgaagcttgt aaaaagggtt ttcagatcgt tagcatttca 6241 cagaatgctc atgaatcctt gttagatccg gccgatacat cattagagtt tgcactaagg 6301 gatactttga aaccaatggc aatctatcgt gtgggaaata tgtttttgct gtgttacaca 6361 gaatttgcct tctttgtcaa taatcagggt tggagaaaga aagaatcgca tattattcat 6421 tgggagggag agccccaaaa atttgcaatt tggtacccgt acatcttggc attcgacagc 6481 aacttcattg agatcagaaa gattgaaact ggcgaattaa ttcgttgcgt ccttgctgat 6541 aagattcgac ttctacaaac aagtactcaa gaaat //