LOCUS Exported 4773 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Tag protein of interest with PIF6 in Saccharomyces cerevisiae. ACCESSION . VERSION . KEYWORDS linker-mCitrine-PIF6-NAT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4773) AUTHORS Yang X, Jost AP, Weiner OD, Tang C TITLE A light-inducible organelle-targeting system for dynamically activating and inactivating signaling in budding yeast. JOURNAL Mol Biol Cell. 2013 Aug;24(15):2419-30. doi: 10.1091/mbc.E13-03-0126. Epub 2013 Jun 12. PUBMED 23761071 REFERENCE 2 (bases 1 to 4773) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4773 /organism="synthetic DNA construct" /mol_type="other DNA" gene 115..1234 /label=natMX6 /note="yeast selectable marker conferring nourseothricin resistance" promoter 115..458 /label=TEF promoter /note="Ashbya gossypii TEF promoter" CDS 459..1031 /codon_start=1 /gene="Streptomyces noursei nat1" /product="nourseothricin acetyltransferase" /label=NrsR /note="confers resistance to nourseothricin" /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT LCGLDTALYDGTASDGEQALYMSMPCP" terminator 1037..1234 /label=TEF terminator /note="Ashbya gossypii TEF terminator" primer_bind complement(2410..2429) /label=T7 /note="T7 promoter, forward primer" promoter complement(2411..2429) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2516..2533) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2687..3275) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2767..2786) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3446..4306) /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 4069..4088 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4307..4411) /gene="bla" /label=AmpR promoter primer_bind 4479..4497 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(4535..4557) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 4657..4676 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 4757..2 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 4757..1 /label=SP6 /note="SP6 promoter, forward primer" ORIGIN 1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta 61 aggcgcgcca gatctgttta gcttgcctcg tccccgccgg gtcacccggc cagcgacatg 121 gaggcccaga ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac 181 tgtcgcccgt acatttagcc catacatccc catgtataat catttgcatc catacatttt 241 gatggccgca cggcgcgaag caaaaattac ggctcctcgc tgcagacctg cgagcaggga 301 aacgctcccc tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata 361 aaaggttagg atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta 421 ggatacagtt ctcacatcac atccgaacat aaacaaccat gggtaccact cttgacgaca 481 cggcttaccg gtaccgcacc agtgtcccgg gggacgccga ggccatcgag gcactggatg 541 ggtccttcac caccgacacc gtcttccgcg tcaccgccac cggggacggc ttcaccctgc 601 gggaggtgcc ggtggacccg cccctgacca aggtgttccc cgacgacgaa tcggacgacg 661 aatcggacga cggggaggac ggcgacccgg actcccggac gttcgtcgcg tacggggacg 721 acggcgacct ggcgggcttc gtggtcgtct cgtactccgg ctggaaccgc cggctgaccg 781 tcgaggacat cgaggtcgcc ccggagcacc gggggcacgg ggtcgggcgc gcgttgatgg 841 ggctcgcgac ggagttcgcc cgcgagcggg gcgccgggca cctctggctg gaggtcacca 901 acgtcaacgc accggcgatc cacgcgtacc ggcggatggg gttcaccctc tgcggcctgg 961 acaccgccct gtacgacggc accgcctcgg acggcgagca ggcgctctac atgagcatgc 1021 cctgccccta atcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt 1081 atagtttttt tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt 1141 ttttcgcctc gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg 1201 cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg attcgatact aacgccgcca 1261 tccagtgtcg aaaacgagct cgaattctta gtcaacatgt ttattgcttt ccaacatgtt 1321 tgtttcatgg gcagcagcaa catgactttg tggaacaacc tgcgtgtccc cgagatttgt 1381 gatggcacca ccaccaccat ggatgatact cttcatgaaa tcctcgttat actctgcctc 1441 atacaaatcc atgatcgatt tggtacgttg attatgcaga gaaacgttgg atctttggcc 1501 ctttgcaaga atctggccat tctcaaacac aagctccata tactcttgat cgcttaacct 1561 gcaacaataa tcggttggta agaacatcat atcgctactc cctgagccgg atccgctccc 1621 tgaacctagg gaactacctt tgtacaattc atccatacca tgggtaatac cagcagcagt 1681 aacaaattct aacaagacca tgtggtctct cttttcgttt ggatctttgg ataatctaga 1741 ttgataggat aagtaatggt tgtctggtaa caagactgga ccatcaccaa ttggagtatt 1801 ttgttgataa tggtcagcta attgaacaga accatcttca atgttgtgtc taattttgaa 1861 gttaactttg ataccattct tttgtttgtc agccatgatg taaacattgt gagagttata 1921 gttgtattcc aatttgtgac ctaaaatgtt accatcttct ttaaaatcaa taccttttaa 1981 ttcgattcta ttaactaagg tatcaccttc aaacttgact tcagctctgg tcttgtagtt 2041 accgtcatct ttgaaaaaaa tagttctttc ttgaacataa ccttctggca tggcagactt 2101 gaaaaagtca tgttgtttca tatgatctgg gtatctagca aaacacatca aaccataacc 2161 taaagtagtg actaaggttg gccatggaac tggcaattta ccagtagtac aaataaattt 2221 taaggtcaat ttaccgtaag tagcatcacc ttcaccttca ccggagacag aaaatttgtg 2281 accattaaca tcaccatcta attcaaccaa aattgggaca acaccagtga ataattcttc 2341 acctttagac attgctttgt taattaaacc agcaccgtca cctgcggcgg cccgcggatc 2401 tgccggtctc cctatagtga gtcgtattaa tttcgataag ccaggttaac ctgcattaat 2461 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc 2521 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 2581 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 2641 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 2701 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 2761 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 2821 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 2881 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 2941 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 3001 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 3061 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 3121 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 3181 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca 3241 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 3301 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa 3361 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta 3421 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag 3481 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga 3541 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac 3601 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc 3661 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta 3721 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac 3781 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat 3841 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa 3901 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg 3961 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag 4021 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc 4081 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct 4141 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat 4201 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg 4261 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc 4321 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta 4381 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg 4441 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct 4501 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga 4561 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag 4621 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga 4681 gagtgcacca tatggacata ttgtcgttag aacgcggcta caattaatac ataaccttat 4741 gtatcataca catacgattt aggtgacact ata //