LOCUS Exported 7047 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION OsTIR1 expression cassette marked with URA. ACCESSION . VERSION . KEYWORDS pJW1661 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7047) AUTHORS Costa EA, Subramanian K, Nunnari J, Weissman JS TITLE Defining the physiological role of SRP in protein-targeting efficiency and specificity. JOURNAL Science. 2018 Jan 18. pii: science.aar3607. doi: 10.1126/science.aar3607. PUBMED 29348368 REFERENCE 2 (bases 1 to 7047) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7047 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(29..51) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 151..170 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 195..410 /gene="S. cerevisiae URA3" /label=URA3 promoter CDS 411..1214 /codon_start=1 /gene="S. cerevisiae URA3" /product="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" /label=URA3 /note="yeast auxotrophic marker, counterselectable with 5-fluoroorotic acid (5-FOA)" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" rep_origin complement(1343..1798) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1480..1501) /label=F1ori-F /note="F1 origin, forward primer" CDS complement(1581..2156) /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSSELTLTKGNKSWVPGPPSRSTVSISLISNSCSPGDPLV LERPPPRWSSNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART DRPSQQLRSLNGEWRDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFAG FPRQALNRGLPLGFRFSALRHLDPKKLD" primer_bind 1692..1711 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1928..1950 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 1942..1959 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 1943..1959 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 1966..1985 /label=T7 /note="T7 promoter, forward primer" promoter 1966..1984 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 1993..2100 /label=MCS /note="pBluescript multiple cloning site" primer_bind 2017..2033 /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind 2017..2033 /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2067..2083) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2068..2084) /label=pBluescriptKS /note="For pBluescript vector" promoter complement(2113..2131) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2152..2168) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2152..2168) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2165..2187) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2176..2192 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(2200..2230) /label=lac promoter /note="promoter for the E. coli lac operon" primer_bind complement(2383..2400) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2554..3142) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3313..4173) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 3936..3955 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4174..4278) /gene="bla" /label=AmpR promoter primer_bind 4346..4364 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4665..5061 /gene="S. cerevisiae ADH1" /label=ADH1 promoter /note="promoter for alcohol dehydrogenase 1" CDS 5074..6801 /codon_start=1 /product="rice transport inhibitor response 1 protein" /label=OsTIR1 /note="F-box protein that mediates auxin-dependent interaction of a ubiquitin ligase with an auxin-inducible degron (Tan et al., 2007)" /translation="MTYFPEEVVEHIFSFLPAQRDRNTVSLVCKVWYEIERLSRRGVFV GNCYAVRAGRVAARFPNVRALTVKGKPHFADFNLVPPDWGGYAGPWIEAAARGCHGLEE LRMKRMVVSDESLELLARSFPRFRALVLISCEGFSTDGLAAVASHCKLLRELDLQENEV EDRGPRWLSCFPDSCTSLVSLNFACIKGEVNAGSLERLVSRSPNLRSLRLNRSVSVDTL AKILLRTPNLEDLGTGNLTDDFQTESYFKLTSALEKCKMLRSLSGFWDASPVCLSFIYP LCAQLTGLNLSYAPTLDASDLTKMISRCVKLQRLWVLDCISDKGLQVVASSCKDLQELR VFPSDFYVAGYSAVTEEGLVAVSLGCPKLNSLLYFCHQMTNAALVTVAKNCPNFTRFRL CILEPGKPDVVTSQPLDEGFGAIVRECKGLQRLSISGLLTDKVFMYIGKYAKQLEMLSI AFAGDSDKGMMHVMNGCKNLRKLEIRDSPFGDAALLGNFARYETMRSLWMSSCNVTLKG CQVLASKMPMLNVEVINERDGSNEMEENHGDLPKVEKLYVYRTTAGARDDAPNFVKIL" terminator 6858..7045 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind complement(6913..6937) /label=pHybLex-R /note="ADH terminator, reverse primer" ORIGIN 1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 181 accacgcttt tcaattcaat tcatcatttt ttttttattc ttttttttga tttcggtttc 241 tttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg aaggagcaca 301 gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat tgcccagtat 361 tcttaaccca actgcacaga acaaaaacct gcaggaaacg aagataaatc atgtcgaaag 421 ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag ctatttaata 481 tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc accaaggaat 541 tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca catgtggata 601 tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta tccgccaagt 661 acaatttttt actcttcgaa gacagaaaat ttgctgacat tggtaataca gtcaaattgc 721 agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat gcacacggtg 781 tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta acaaaggaac 841 ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct actggagaat 901 atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc ggctttattg 961 ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg acacccggtg 1021 tgggtttaga tgacaaggga gacgcattgg gtcaacagta tagaaccgtg gatgatgtgg 1081 tctctacagg atctgacatt attattgttg gaagaggact atttgcaaag ggaagggatg 1141 ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg agaagatgcg 1201 gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac tcacaaatta 1261 gagcttcaat ttaattatat cagttattac cctgcggtgt gaaataccgc acagatgcgt 1321 aaggagaaaa taccgcatca ggaaattgta aacgttaata ttttgttaaa attcgcgtta 1381 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat 1441 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca 1501 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc 1561 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta 1621 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg 1681 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg 1741 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg 1801 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg 1861 ctattacgcc agctggcgaa ggggggatgt gctgcaaggc gattaagttg ggtaacgcca 1921 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattgtaata cgactcacta 1981 tagggcgaat tggagctcca ccgcggtggc ggccgctcta gaactagtgg atcccccggg 2041 ctgcaggaat tcgatatcaa gcttatcgat accgtcgacc tcgagggggg gcccggtacc 2101 cagcttttgt tccctttagt gagggttaat tccgagcttg gcgtaatcat ggtcatagct 2161 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacataggag ccggaagcat 2221 aaagtgtaaa gcctggggtg cctaatgagt gaggtaactc acattaattg cgttgcgctc 2281 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 2341 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 2401 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 2461 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 2521 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctcggcc cccctgacga 2581 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 2641 ccaggcgttc ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 2701 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg 2761 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 2821 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 2881 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 2941 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt 3001 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 3061 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 3121 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 3181 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 3241 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 3301 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 3361 tcgttcatcc atagttgcct gactgcccgt cgtgtagata actacgatac gggagggctt 3421 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 3481 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 3541 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 3601 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 3661 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 3721 gtgaaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 3781 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 3841 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 3901 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 3961 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 4021 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 4081 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 4141 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 4201 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 4261 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat 4321 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtcgggtgta 4381 caatatggac ttcctctttt ctggcaacca aacccataca tcgggattcc tataatacct 4441 tcgttggtct ccctaacatg taggtggcgg aggggagata tacaatagaa cagataccag 4501 acaagacata atgggctaaa caagactaca ccaattacac tgcctcattg atggtggtac 4561 ataacgaact aatactgtag ccctagactt gatagccatc atcatatcga agtttcacta 4621 ccctttttcc atttgccatc tattgaagta ataataggcg catgcaactt cttttctttt 4681 tttttctttt ctctctcccc cgttgttgtc tcaccatatc cgcaatgaca aaaaaatgat 4741 ggaagacact aaaggaaaaa attaacgaca aagacagcac caacagatgt cgttgttcca 4801 gagctgatga ggggtatctc gaagcacacg aaactttttc cttccttcat tcacgcacac 4861 tactctctaa tgagcaacgg tatacggcct tccttccagt tacttgaatt tgaaataaaa 4921 aaaagtttgc tgtcttgcta tcaagtataa atagacctgc aattattaat cttttgtttc 4981 ctcgtcattg ttctcgttcc ctttcttcct tgtttctttt tctgcacaat atttcaagct 5041 ataccaagca tacaatcaac tatctcatat acaatgacgt acttcccgga ggaggtggtg 5101 gagcacatct tcagcttcct gccggcgcag cgcgaccgca acacggtctc gctcgtctgc 5161 aaggtgtggt acgagatcga gaggctgagc cgccgcggcg tcttcgtggg caactgctac 5221 gccgtgcgcg ccggccgcgt cgccgcgcgg ttccccaacg tgcgggcgct cacggtgaag 5281 gggaagcccc acttcgccga cttcaacctc gtgccccccg actggggcgg ctacgcgggg 5341 ccgtggatcg aggcggccgc gaggggatgc cacggcctgg aggagctcag gatgaagcgg 5401 atggtggtgt ccgacgagag cctcgagctg ctggctcgct cgttcccgcg gttcagggct 5461 cttgttctta tcagctgcga ggggttcagc actgacgggc tagccgccgt cgcgagccat 5521 tgcaagcttc tgagggagtt ggatttgcag gaaaatgaag tggaggatcg agggcctagg 5581 tggctttcct gcttccctga ttcctgcaca tcacttgtct cattgaattt tgcctgcatc 5641 aaaggggagg ttaatgctgg ttcactggag agacttgtta gcaggtcccc aaacctgcgg 5701 agtttgaggc tgaatcgatc tgtatcggta gatacacttg caaagatact actgcgtacc 5761 cctaacttgg aggatttggg gacagggaat ttgacagatg acttccaaac tgagtcctac 5821 tttaagctta ccagtgctct ggagaaatgc aagatgttga ggagtttgtc tggattctgg 5881 gatgcttctc ctgtttgcct gtcatttatc taccccctgt gtgctcaact gacaggattg 5941 aacttgagct atgcccccac acttgatgct tctgacctta caaaaatgat tagccgctgt 6001 gtgaagctcc aacgcctttg ggtactggat tgtatctcgg acaaaggctt gcaagtggtg 6061 gcctccagtt gcaaagactt gcaagaactc agggtatttc catcagattt ctacgtagct 6121 ggttattctg cagtgacaga ggagggactt gttgcagtat ccttgggctg tccaaaactg 6181 aactcactac tgtacttctg tcaccaaatg actaatgctg cactagttac tgtcgccaag 6241 aactgtccaa atttcacacg attcagactt tgtattcttg agccagggaa gcctgatgtt 6301 gtgacaagcc aaccattaga tgaaggcttt ggagctattg ttcgtgagtg caagggatta 6361 caacgtttgt caatatctgg tcttctcaca gacaaagttt tcatgtatat tgggaaatat 6421 gcaaaacaac ttgagatgct ttctatagca tttgctggtg acagtgataa gggtatgatg 6481 catgttatga atggatgcaa gaatttaagg aaactggaga taagagatag cccgtttggt 6541 gatgctgcac tcttggggaa ttttgctagg tacgagacaa tgcgatccct ttggatgtca 6601 tcttgcaatg tcacgttaaa ggggtgccaa gtccttgcgt caaagatgcc gatgctcaat 6661 gttgaggtca taaatgagcg ggatggtagc aatgaaatgg aggaaaacca tggagatctg 6721 cctaaagtgg agaaattata tgtgtaccgc acaactgctg gggcgaggga tgatgcacca 6781 aattttgtta aaatcctatg ataggcctat ccctaaccct ctcctcggtc tcgattctac 6841 gtaacacttc taaataagcg aatttcttat gatttatgat ttttattatt aaataagtta 6901 taaaaaaaat aagtgtatac aaattttaaa gtgactctta ggttttaaaa cgaaaattct 6961 tattcttgag taactctttc ctgtaggtca ggttgctttc tcaggtatag catgaggtcg 7021 ctcttattga ccacacctct accggca //