LOCUS Exported 8180 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Integrative expression of yEGFP-AID*-CUP1 and rice auxin receptor gene OsTIR1. . ACCESSION . VERSION . KEYWORDS pJAID9L104 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8180) TITLE Vickers lab yeast integrative expression plasmids REFERENCE 2 (bases 1 to 8180) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8180 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..408 /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" CDS 431..1141 /codon_start=1 /product="yeast-enhanced green fluorescent protein" /label=yeGFP /note="codons optimized for translation in S. cerevisiae and C. albicans" /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF VTAAGITHGMDELYK" terminator 1472..1742 /gene="S. cerevisae PGK1" /label=PGK1 terminator /note="transcription terminator for 3-phosphoglycerate kinase gene" CDS 2238..3965 /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" primer_bind complement(4201..4217) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4201..4217) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4214..4236) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4225..4241 /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(4249..4279) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4294..4315 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4432..4449) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4603..5191) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4683..4702) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5362..6222) /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 5985..6004 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6223..6327) /gene="bla" /label=AmpR promoter primer_bind 6395..6413 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(6451..6473) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 6573..6592 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 6786..6808 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 6800..6817 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 6801..6817 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 7931..7964 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." ORIGIN 1 ccacacacca tagcttcaaa atgtttctac tcctttttta ctcttccaga ttttctcgga 61 ctccgcgcat cgccgtacca cttcaaaaca cccaagcaca gcatactaaa tttcccctct 121 ttcttcctct agggtgtcgt taattacccg tactaaaggt ttggaaaaga aaaaagagac 181 cgcctcgttt ctttttcttc gtcgaaaaag gcaataaaaa tttttatcac gtttcttttt 241 cttgaaaatt tttttttttg atttttttct ctttcgatga cctcccattg atatttaagt 301 taataaacgg tcttcaattt ctcaagtttc agtttcattt ttcttgttct attacaactt 361 tttttacttc ttgctcatta gaaagaaagc atagcaatct aatctaagtt ttaattacaa 421 aatgggatcc tctaaaggtg aagaattatt cactggtgtt gtcccaattt tggttgaatt 481 agatggtgat gttaatggtc acaaattttc tgtctccggt gaaggtgaag gtgatgctac 541 ttacggtaaa ttgaccttaa aatttatttg tactactggt aaattgccag ttccatggcc 601 aaccttagtc actactttcg gttatggtgt tcaatgtttt gcgagatacc cagatcatat 661 gaaacaacat gactttttca agtctgccat gccagaaggt tatgttcaag aaagaactat 721 ttttttcaaa gatgacggta actacaagac cagagctgaa gtcaagtttg aaggtgatac 781 cttagttaat agaatcgaat taaaaggtat tgattttaaa gaagatggta acattttagg 841 tcacaaattg gaatacaact ataactctca caatgtttac atcatggctg acaaacaaaa 901 gaatggtatc aaagttaact tcaaaattag acacaacatt gaagatggtt ctgttcaatt 961 agctgaccat tatcaacaaa atactccaat tggtgatggt ccagtcttgt taccagacaa 1021 ccattactta tccactcaat ctgccttatc caaagatcca aacgaaaaga gagaccacat 1081 ggtcttgtta gaatttgtta ctgctgctgg tattacccat ggtatggatg aattgtacaa 1141 aagatctcca aaagacccag ctaaacctcc agccaaggcc caagtcgtcg gctggccacc 1201 agttaggtct tacagaaaga acgtaatggt tagttgtcaa aagtcctccg gtggtccaga 1261 agctgctgcc tttgttaaga tgttcagcga attaattaac ttccaaaatg aaggtcatga 1321 gtgccaatgc caatgtggta gctgcaaaaa taatgaacaa tgccaaaaat catgtagctg 1381 cccaacgggg tgtaacagcg acgacaaatg cccctgcggt aacaagtctg aagaaaccaa 1441 gaagtcatgc tgctctggga aataaactag tattgaattg aattgaaatc gatagatcaa 1501 tttttttctt ttctctttcc ccatccttta cgctaaaata atagtttatt ttattttttg 1561 aatatttttt atttatatac gtatatatag actattattt atcttttaat gattattaag 1621 atttttatta aaaaaaaatt cgctcctctt ttaatgcctt tatgcagttt ttttttccca 1681 ttcgatattt ctatgttcgg gttcagcgta ttttaagttt aataactcga aaattctgcg 1741 ttcgttaaag ctttcgagaa ggatattatt tgcggccgcg attgcaagaa ttgatctccc 1801 tcatcgatgt cataaatcag taaatataaa cattaaatac attacacgta gccacccgct 1861 gtatagcata aaactaactt aaaattattc atatgcgttt cccggggccg aagcgttatt 1921 gccgatattc ggtagccgat tccgctggct tttttttttt gaacgaggca aaaaagtttc 1981 gtgactgaac ttcttcgtca ctggttttgt tgataatttt tatcgttttt tacttcgaga 2041 ttgaaaacag tgttctgaat tttagcaacg atatcgcacc gcacgtttag cctgatcaaa 2101 ccttatatat acctctacct taaaatatat atatatatag tattcgaatt ccgaattgtg 2161 tgtctccgcg gttagtgatt gttatacaca aacagaatac aggaaagtaa atcaatacaa 2221 taataaaagc tagcaaaatg acctatttcc cagaagaagt cgttgaacac atcttctctt 2281 tcttgccagc tcaaagagat agaaatactg tttccttggt ttgtaaggtt tggtatgaaa 2341 tcgaaagatt gagtagacgt ggtgttttcg tcggtaattg ttacgctgtt agagctggta 2401 gagttgctgc tagattccca aatgttagag ccttgactgt caaaggtaaa ccacattttg 2461 ctgactttaa cttggtccca ccagattggg gtggttatgc tggtccatgg attgaagctg 2521 ccgctagagg ttgtcacggt ttggaagaat tgcgtatgaa aagaatggtc gtttctgacg 2581 aatctttgga attattggcc cgttcttttc caagattcag agctttggtt ttgatctctt 2641 gtgaaggttt cagtactgac ggtttggctg ccgttgcttc ccattgtaaa ttattgagag 2701 aattggactt gcaagaaaat gaagtcgaag atagaggtcc aagatggtta tcttgttttc 2761 cagactcttg tacttctttg gtctctttga acttcgcttg tattaagggt gaagtcaacg 2821 ctggttcttt agaaagattg gtctcccgtt ctccaaactt acgttctttg agattgaatc 2881 gttctgtttc cgttgatacc ttggctaaaa tcttattgag aaccccaaac ttggaagact 2941 taggtactgg taacttgacc gacgatttcc aaactgaatc ctactttaaa ttaacttctg 3001 ctttagaaaa gtgtaagatg ttgagatcct tatctggttt ttgggacgcc tctccagtct 3061 gtttatcttt tatctatcca ttgtgtgctc aattaactgg tttaaactta tcttacgctc 3121 caaccttgga tgctagtgac ttaaccaaga tgattagtag atgtgttaag ttacaaagat 3181 tatgggtttt agattgtatt tccgataagg gtttacaagt cgttgctagt agctgtaagg 3241 atttgcaaga attaagagtc tttccaagtg atttttacgt tgctggttac agcgctgtta 3301 ctgaagaagg tttggtcgct gtttctttgg gttgtccaaa gttgaacagc ttattgtact 3361 tttgtcatca aatgactaat gctgccttag tcactgttgc taagaactgt ccaaacttca 3421 cccgtttcag attgtgtatt ttggaaccag gtaaaccaga cgttgtcacc tcccaaccat 3481 tggatgaagg tttcggtgct atcgttagag aatgtaaggg tttgcaaaga ttatctatct 3541 ctggtttgtt aactgacaaa gttttcatgt acatcggtaa atatgccaaa caattagaaa 3601 tgttgagcat tgctttcgct ggtgactctg acaagggtat gatgcacgtt atgaatggtt 3661 gtaagaattt gagaaagttg gaaattagag attctccatt cggtgacgct gccttattgg 3721 gtaattttgc tagatacgaa actatgagaa gtttgtggat gagttcctgt aacgttacct 3781 tgaaaggttg tcaagtctta gccagcaaga tgccaatgtt gaacgtcgaa gtcatcaacg 3841 aaagagacgg tagtaacgaa atggaagaaa atcacggtga cttaccaaag gtagaaaagt 3901 tgtacgtata cagaactact gctggtgcta gagatgatgc tccaaatttc gtaaagattt 3961 tgtaactcga gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac 4021 agaaaagcag gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta 4081 ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta attatatcag 4141 ttattacccg ggaatctcgg tcgtaatgat ttaaatgcat gcaagcttgg cgtaatcatg 4201 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 4261 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 4321 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 4381 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 4441 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 4501 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 4561 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 4621 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 4681 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 4741 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 4801 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 4861 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 4921 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 4981 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 5041 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 5101 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 5161 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 5221 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 5281 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 5341 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 5401 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 5461 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 5521 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 5581 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 5641 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 5701 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 5761 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 5821 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 5881 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 5941 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 6001 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 6061 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 6121 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 6181 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 6241 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 6301 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta 6361 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg 6421 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt 6481 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg 6541 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt 6601 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg 6661 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct 6721 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg 6781 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttcatttaaa tttttttgat 6841 tcggtaatct ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat 6901 atacgcatat gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac 6961 agaacaaaaa cctgcaggaa acgaagataa aacatgtcca caaaatcata taccagtaga 7021 gctgagactc atgcaagtcc ggttgcatcg aaacttttac gtttaatgga tgaaaagaag 7081 accaatttgt gtgcttctct tgacgttcgt tcgactgatg agctattgaa acttgttgaa 7141 acgttgggtc catacatttg ccttttgaaa acacacgttg atatcttgga tgatttcagt 7201 tatgagggta ctgtcgttcc attgaaagca ttggcagaga aatacaagtt cttgatattt 7261 gaggacagaa aattcgccga tatcggtaac acagtcaaat tacaatatac atcgggcgtt 7321 taccgtatcg cagaatggtc tgatatcacc aacgcccacg gggttactgg tgctggtatt 7381 gttgctggct tgaaacaagg tgcgcaagag gtcaccaaag aaccaagggg attattgatg 7441 cttgctgaat tgtcttccaa gggttctcta gcacacggtg aatatactaa gggtaccgtt 7501 gatattgcaa agagtgataa agatttcgtt attgggttca ttgctcagaa cgatatggga 7561 ggaagagaag aagggtttga ttggctaatc atgaccccag gtgtaggttt agacgacaaa 7621 ggcgatgcat tgggtcagca gtacagaacc gtcgacgaag ttgtaagtgg tggatcagat 7681 atcatcattg ttggcagagg acttttcgcc aagggtagag atcctaaggt tgaaggtgaa 7741 agatacagaa atgctggatg ggaagcgtac caaaagagaa tcagcgctcc ccattaatta 7801 tacaggaaac ttaatagaac aaatcacata tttaatctaa tagccacctg cattggcacg 7861 gtgcaacact acttcaactt catcttacaa aaagatcacg tgatctgttg tattgggatc 7921 tctagaccta ataacttcgt atagcataca ttatacgaag ttatattaag ggttgctcga 7981 ccagaaagcg accacccaac tttggctgat aatagcgtat aaacaatgca tactttgtac 8041 gttcaaaata caatgcagta gatatattta tgcatattac atataataca tatcacatag 8101 gaagcaacag gcgcgttgga cttttaattt tcgaggaccg cgaatcctta catcacaccc 8161 aatcccccac aagtgatccc //