LOCUS Exported 7868 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Integrative expression of yEGFP and rice auxin receptor gene OsTIR1. OsTIR1 is codon-optimised for expression in S. cerevisiae. ACCESSION . VERSION . KEYWORDS pJAID6A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7868) TITLE Vickers lab yeast integrative expression plasmids REFERENCE 2 (bases 1 to 7868) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7868 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 9..1736 /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(1966..1982) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1966..1982) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1979..2001) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1990..2006 /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(2014..2044) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2059..2080 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2197..2214) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2368..2956) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2448..2467) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3127..3987) /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 3750..3769 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3988..4092) /gene="bla" /label=AmpR promoter primer_bind 4160..4178 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(4216..4238) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 4338..4357 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4551..4573 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 4565..4582 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 4566..4582 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 5696..5729 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." promoter 5946..6353 /gene="S. cerevisiae TEF1" /label=TEF1 promoter /note="promoter for EF-1-alpha" CDS 6385..7095 /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 7111..7381 /gene="S. cerevisae PGK1" /label=PGK1 terminator /note="transcription terminator for 3-phosphoglycerate kinase gene" ORIGIN 1 ctagcaaaat gacctatttc ccagaagaag tcgttgaaca catcttctct ttcttgccag 61 ctcaaagaga tagaaatact gtttccttgg tttgtaaggt ttggtatgaa atcgaaagat 121 tgagtagacg tggtgttttc gtcggtaatt gttacgctgt tagagctggt agagttgctg 181 ctagattccc aaatgttaga gccttgactg tcaaaggtaa accacatttt gctgacttta 241 acttggtccc accagattgg ggtggttatg ctggtccatg gattgaagct gccgctagag 301 gttgtcacgg tttggaagaa ttgcgtatga aaagaatggt cgtttctgac gaatctttgg 361 aattattggc ccgttctttt ccaagattca gagctttggt tttgatctct tgtgaaggtt 421 tcagtactga cggtttggct gccgttgctt cccattgtaa attattgaga gaattggact 481 tgcaagaaaa tgaagtcgaa gatagaggtc caagatggtt atcttgtttt ccagactctt 541 gtacttcttt ggtctctttg aacttcgctt gtattaaggg tgaagtcaac gctggttctt 601 tagaaagatt ggtctcccgt tctccaaact tacgttcttt gagattgaat cgttctgttt 661 ccgttgatac cttggctaaa atcttattga gaaccccaaa cttggaagac ttaggtactg 721 gtaacttgac cgacgatttc caaactgaat cctactttaa attaacttct gctttagaaa 781 agtgtaagat gttgagatcc ttatctggtt tttgggacgc ctctccagtc tgtttatctt 841 ttatctatcc attgtgtgct caattaactg gtttaaactt atcttacgct ccaaccttgg 901 atgctagtga cttaaccaag atgattagta gatgtgttaa gttacaaaga ttatgggttt 961 tagattgtat ttccgataag ggtttacaag tcgttgctag tagctgtaag gatttgcaag 1021 aattaagagt ctttccaagt gatttttacg ttgctggtta cagcgctgtt actgaagaag 1081 gtttggtcgc tgtttctttg ggttgtccaa agttgaacag cttattgtac ttttgtcatc 1141 aaatgactaa tgctgcctta gtcactgttg ctaagaactg tccaaacttc acccgtttca 1201 gattgtgtat tttggaacca ggtaaaccag acgttgtcac ctcccaacca ttggatgaag 1261 gtttcggtgc tatcgttaga gaatgtaagg gtttgcaaag attatctatc tctggtttgt 1321 taactgacaa agttttcatg tacatcggta aatatgccaa acaattagaa atgttgagca 1381 ttgctttcgc tggtgactct gacaagggta tgatgcacgt tatgaatggt tgtaagaatt 1441 tgagaaagtt ggaaattaga gattctccat tcggtgacgc tgccttattg ggtaattttg 1501 ctagatacga aactatgaga agtttgtgga tgagttcctg taacgttacc ttgaaaggtt 1561 gtcaagtctt agccagcaag atgccaatgt tgaacgtcga agtcatcaac gaaagagacg 1621 gtagtaacga aatggaagaa aatcacggtg acttaccaaa ggtagaaaag ttgtacgtat 1681 acagaactac tgctggtgct agagatgatg ctccaaattt cgtaaagatt ttgtaaagga 1741 ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg 1801 gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca 1861 tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt acccgggaat 1921 ctcggtcgta atgatttaaa tgcatgcaag cttggcgtaa tcatggtcat agctgtttcc 1981 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg 2041 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc 2101 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 2161 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 2221 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 2281 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 2341 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 2401 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 2461 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 2521 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 2581 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 2641 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 2701 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 2761 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg 2821 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 2881 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 2941 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 3001 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 3061 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 3121 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 3181 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 3241 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 3301 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 3361 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 3421 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 3481 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 3541 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 3601 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 3661 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 3721 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 3781 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 3841 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 3901 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 3961 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 4021 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 4081 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat 4141 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg 4201 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta 4261 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg 4321 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg 4381 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag 4441 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc 4501 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 4561 acgttgtaaa acgacggcca gtgaattcat ttaaattttt ttgattcggt aatctccgaa 4621 cagaaggaag aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt 4681 gttgaagaaa catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacctgc 4741 aggaaacgaa gataaaacat gtccacaaaa tcatatacca gtagagctga gactcatgca 4801 agtccggttg catcgaaact tttacgttta atggatgaaa agaagaccaa tttgtgtgct 4861 tctcttgacg ttcgttcgac tgatgagcta ttgaaacttg ttgaaacgtt gggtccatac 4921 atttgccttt tgaaaacaca cgttgatatc ttggatgatt tcagttatga gggtactgtc 4981 gttccattga aagcattggc agagaaatac aagttcttga tatttgagga cagaaaattc 5041 gccgatatcg gtaacacagt caaattacaa tatacatcgg gcgtttaccg tatcgcagaa 5101 tggtctgata tcaccaacgc ccacggggtt actggtgctg gtattgttgc tggcttgaaa 5161 caaggtgcgc aagaggtcac caaagaacca aggggattat tgatgcttgc tgaattgtct 5221 tccaagggtt ctctagcaca cggtgaatat actaagggta ccgttgatat tgcaaagagt 5281 gataaagatt tcgttattgg gttcattgct cagaacgata tgggaggaag agaagaaggg 5341 tttgattggc taatcatgac cccaggtgta ggtttagacg acaaaggcga tgcattgggt 5401 cagcagtaca gaaccgtcga cgaagttgta agtggtggat cagatatcat cattgttggc 5461 agaggacttt tcgccaaggg tagagatcct aaggttgaag gtgaaagata cagaaatgct 5521 ggatgggaag cgtaccaaaa gagaatcagc gctccccatt aattatacag gaaacttaat 5581 agaacaaatc acatatttaa tctaatagcc acctgcattg gcacggtgca acactacttc 5641 aacttcatct tacaaaaaga tcacgtgatc tgttgtattg ggatctctag acctaataac 5701 ttcgtatagc atacattata cgaagttata ttaagggttg ctcgaccaga aagcgaccac 5761 ccaactttgg ctgataatag cgtataaaca atgcatactt tgtacgttca aaatacaatg 5821 cagtagatat atttatgcat attacatata atacatatca cataggaagc aacaggcgcg 5881 ttggactttt aattttcgag gaccgcgaat ccttacatca cacccaatcc cccacaagtg 5941 atcccccaca caccatagct tcaaaatgtt tctactcctt ttttactctt ccagattttc 6001 tcggactccg cgcatcgccg taccacttca aaacacccaa gcacagcata ctaaatttcc 6061 cctctttctt cctctagggt gtcgttaatt acccgtacta aaggtttgga aaagaaaaaa 6121 gagaccgcct cgtttctttt tcttcgtcga aaaaggcaat aaaaattttt atcacgtttc 6181 tttttcttga aaattttttt ttttgatttt tttctctttc gatgacctcc cattgatatt 6241 taagttaata aacggtcttc aatttctcaa gtttcagttt catttttctt gttctattac 6301 aacttttttt acttcttgct cattagaaag aaagcatagc aatctaatct aagttttaat 6361 tacaaactcg agaaaatggg atcctctaaa ggtgaagaat tattcactgg tgttgtccca 6421 attttggttg aattagatgg tgatgttaat ggtcacaaat tttctgtctc cggtgaaggt 6481 gaaggtgatg ctacttacgg taaattgacc ttaaaattta tttgtactac tggtaaattg 6541 ccagttccat ggccaacctt agtcactact ttcggttatg gtgttcaatg ttttgcgaga 6601 tacccagatc atatgaaaca acatgacttt ttcaagtctg ccatgccaga aggttatgtt 6661 caagaaagaa ctattttttt caaagatgac ggtaactaca agaccagagc tgaagtcaag 6721 tttgaaggtg ataccttagt taatagaatc gaattaaaag gtattgattt taaagaagat 6781 ggtaacattt taggtcacaa attggaatac aactataact ctcacaatgt ttacatcatg 6841 gctgacaaac aaaagaatgg tatcaaagtt aacttcaaaa ttagacacaa cattgaagat 6901 ggttctgttc aattagctga ccattatcaa caaaatactc caattggtga tggtccagtc 6961 ttgttaccag acaaccatta cttatccact caatctgcct tatccaaaga tccaaacgaa 7021 aagagagacc acatggtctt gttagaattt gttactgctg ctggtattac ccatggtatg 7081 gatgaattgt acaaaagatc ttaaactagt attgaattga attgaaatcg atagatcaat 7141 ttttttcttt tctctttccc catcctttac gctaaaataa tagtttattt tattttttga 7201 atatttttta tttatatacg tatatataga ctattattta tcttttaatg attattaaga 7261 tttttattaa aaaaaaattc gctcctcttt taatgccttt atgcagtttt tttttcccat 7321 tcgatatttc tatgttcggg ttcagcgtat tttaagttta ataactcgaa aattctgcgt 7381 tcgttaaagc tttcgagaag gatattattt gcggccgcga ttgcaagaat tgatctccct 7441 catcgatgtc ataaatcagt aaatataaac attaaataca ttacacgtag ccacccgctg 7501 tatagcataa aactaactta aaattattca tatgcgtttc ccggggccga agcgttattg 7561 ccgatattcg gtagccgatt ccgctggctt tttttttttg aacgaggcaa aaaagtttcg 7621 tgactgaact tcttcgtcac tggttttgtt gataattttt atcgtttttt acttcgagat 7681 tgaaaacagt gttctgaatt ttagcaacga tatcgcaccg cacgtttagc ctgatcaaac 7741 cttatatata cctctacctt aaaatatata tatatatagt attcgaattc cgaattgtgt 7801 gtctccgcgg ttagtgattg ttatacacaa acagaataca ggaaagtaaa tcaatacaat 7861 aataaatt //