LOCUS Exported 8003 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Integrative expression of AID* (mini auxin inducible degron)-yEGFP and rice auxin receptor gene OsTIR1. . ACCESSION . VERSION . KEYWORDS pJAID5Z SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8003) TITLE Vickers lab yeast integrative expression plasmids REFERENCE 2 (bases 1 to 8003) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8003 /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 575..1285 /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 1301..1571 /gene="S. cerevisae PGK1" /label=PGK1 terminator /note="transcription terminator for 3-phosphoglycerate kinase gene" CDS 2067..3794 /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(4024..4040) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4024..4040) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4037..4059) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4048..4064 /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(4072..4102) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4117..4138 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4255..4272) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4426..5014) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4506..4525) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5185..6045) /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 5808..5827 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6046..6150) /gene="bla" /label=AmpR promoter primer_bind 6218..6236 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(6274..6296) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 6396..6415 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 6609..6631 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 6623..6640 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 6624..6640 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 7754..7787 /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 actcgagaaa atgccaaaag acccagctaa acctccagcc aaggcccaag tcgtcggctg 481 gccaccagtt aggtcttaca gaaagaacgt aatggttagt tgtcaaaagt cctccggtgg 541 tccagaagct gctgcctttg ttaagatggg atcctctaaa ggtgaagaat tattcactgg 601 tgttgtccca attttggttg aattagatgg tgatgttaat ggtcacaaat tttctgtctc 661 cggtgaaggt gaaggtgatg ctacttacgg taaattgacc ttaaaattta tttgtactac 721 tggtaaattg ccagttccat ggccaacctt agtcactact ttcggttatg gtgttcaatg 781 ttttgcgaga tacccagatc atatgaaaca acatgacttt ttcaagtctg ccatgccaga 841 aggttatgtt caagaaagaa ctattttttt caaagatgac ggtaactaca agaccagagc 901 tgaagtcaag tttgaaggtg ataccttagt taatagaatc gaattaaaag gtattgattt 961 taaagaagat ggtaacattt taggtcacaa attggaatac aactataact ctcacaatgt 1021 ttacatcatg gctgacaaac aaaagaatgg tatcaaagtt aacttcaaaa ttagacacaa 1081 cattgaagat ggttctgttc aattagctga ccattatcaa caaaatactc caattggtga 1141 tggtccagtc ttgttaccag acaaccatta cttatccact caatctgcct tatccaaaga 1201 tccaaacgaa aagagagacc acatggtctt gttagaattt gttactgctg ctggtattac 1261 ccatggtatg gatgaattgt acaaaagatc ttaaactagt attgaattga attgaaatcg 1321 atagatcaat ttttttcttt tctctttccc catcctttac gctaaaataa tagtttattt 1381 tattttttga atatttttta tttatatacg tatatataga ctattattta tcttttaatg 1441 attattaaga tttttattaa aaaaaaattc gctcctcttt taatgccttt atgcagtttt 1501 tttttcccat tcgatatttc tatgttcggg ttcagcgtat tttaagttta ataactcgaa 1561 aattctgcgt tcgttaaagc tttcgagaag gatattattt gcggccgcga ttgcaagaat 1621 tgatctccct catcgatgtc ataaatcagt aaatataaac attaaataca ttacacgtag 1681 ccacccgctg tatagcataa aactaactta aaattattca tatgcgtttc ccggggccga 1741 agcgttattg ccgatattcg gtagccgatt ccgctggctt tttttttttg aacgaggcaa 1801 aaaagtttcg tgactgaact tcttcgtcac tggttttgtt gataattttt atcgtttttt 1861 acttcgagat tgaaaacagt gttctgaatt ttagcaacga tatcgcaccg cacgtttagc 1921 ctgatcaaac cttatatata cctctacctt aaaatatata tatatatagt attcgaattc 1981 cgaattgtgt gtctccgcgg ttagtgattg ttatacacaa acagaataca ggaaagtaaa 2041 tcaatacaat aataaattct agcaaaatga cctatttccc agaagaagtc gttgaacaca 2101 tcttctcttt cttgccagct caaagagata gaaatactgt ttccttggtt tgtaaggttt 2161 ggtatgaaat cgaaagattg agtagacgtg gtgttttcgt cggtaattgt tacgctgtta 2221 gagctggtag agttgctgct agattcccaa atgttagagc cttgactgtc aaaggtaaac 2281 cacattttgc tgactttaac ttggtcccac cagattgggg tggttatgct ggtccatgga 2341 ttgaagctgc cgctagaggt tgtcacggtt tggaagaatt gcgtatgaaa agaatggtcg 2401 tttctgacga atctttggaa ttattggccc gttcttttcc aagattcaga gctttggttt 2461 tgatctcttg tgaaggtttc agtactgacg gtttggctgc cgttgcttcc cattgtaaat 2521 tattgagaga attggacttg caagaaaatg aagtcgaaga tagaggtcca agatggttat 2581 cttgttttcc agactcttgt acttctttgg tctctttgaa cttcgcttgt attaagggtg 2641 aagtcaacgc tggttcttta gaaagattgg tctcccgttc tccaaactta cgttctttga 2701 gattgaatcg ttctgtttcc gttgatacct tggctaaaat cttattgaga accccaaact 2761 tggaagactt aggtactggt aacttgaccg acgatttcca aactgaatcc tactttaaat 2821 taacttctgc tttagaaaag tgtaagatgt tgagatcctt atctggtttt tgggacgcct 2881 ctccagtctg tttatctttt atctatccat tgtgtgctca attaactggt ttaaacttat 2941 cttacgctcc aaccttggat gctagtgact taaccaagat gattagtaga tgtgttaagt 3001 tacaaagatt atgggtttta gattgtattt ccgataaggg tttacaagtc gttgctagta 3061 gctgtaagga tttgcaagaa ttaagagtct ttccaagtga tttttacgtt gctggttaca 3121 gcgctgttac tgaagaaggt ttggtcgctg tttctttggg ttgtccaaag ttgaacagct 3181 tattgtactt ttgtcatcaa atgactaatg ctgccttagt cactgttgct aagaactgtc 3241 caaacttcac ccgtttcaga ttgtgtattt tggaaccagg taaaccagac gttgtcacct 3301 cccaaccatt ggatgaaggt ttcggtgcta tcgttagaga atgtaagggt ttgcaaagat 3361 tatctatctc tggtttgtta actgacaaag ttttcatgta catcggtaaa tatgccaaac 3421 aattagaaat gttgagcatt gctttcgctg gtgactctga caagggtatg atgcacgtta 3481 tgaatggttg taagaatttg agaaagttgg aaattagaga ttctccattc ggtgacgctg 3541 ccttattggg taattttgct agatacgaaa ctatgagaag tttgtggatg agttcctgta 3601 acgttacctt gaaaggttgt caagtcttag ccagcaagat gccaatgttg aacgtcgaag 3661 tcatcaacga aagagacggt agtaacgaaa tggaagaaaa tcacggtgac ttaccaaagg 3721 tagaaaagtt gtacgtatac agaactactg ctggtgctag agatgatgct ccaaatttcg 3781 taaagatttt gtaaaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt 3841 tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact 3901 gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat 3961 cagttattac ccgggaatct cggtcgtaat gatttaaatg catgcaagct tggcgtaatc 4021 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 4081 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 4141 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 4201 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 4261 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 4321 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 4381 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 4441 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 4501 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 4561 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 4621 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 4681 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 4741 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 4801 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 4861 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 4921 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 4981 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 5041 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 5101 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 5161 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 5221 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 5281 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 5341 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 5401 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 5461 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 5521 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 5581 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 5641 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 5701 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 5761 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 5821 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 5881 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 5941 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 6001 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 6061 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 6121 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt 6181 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt 6241 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac 6301 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc 6361 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag 6421 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag 6481 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc 6541 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc 6601 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcattt aaattttttt 6661 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta 6721 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg 6781 cacagaacaa aaacctgcag gaaacgaaga taaaacatgt ccacaaaatc atataccagt 6841 agagctgaga ctcatgcaag tccggttgca tcgaaacttt tacgtttaat ggatgaaaag 6901 aagaccaatt tgtgtgcttc tcttgacgtt cgttcgactg atgagctatt gaaacttgtt 6961 gaaacgttgg gtccatacat ttgccttttg aaaacacacg ttgatatctt ggatgatttc 7021 agttatgagg gtactgtcgt tccattgaaa gcattggcag agaaatacaa gttcttgata 7081 tttgaggaca gaaaattcgc cgatatcggt aacacagtca aattacaata tacatcgggc 7141 gtttaccgta tcgcagaatg gtctgatatc accaacgccc acggggttac tggtgctggt 7201 attgttgctg gcttgaaaca aggtgcgcaa gaggtcacca aagaaccaag gggattattg 7261 atgcttgctg aattgtcttc caagggttct ctagcacacg gtgaatatac taagggtacc 7321 gttgatattg caaagagtga taaagatttc gttattgggt tcattgctca gaacgatatg 7381 ggaggaagag aagaagggtt tgattggcta atcatgaccc caggtgtagg tttagacgac 7441 aaaggcgatg cattgggtca gcagtacaga accgtcgacg aagttgtaag tggtggatca 7501 gatatcatca ttgttggcag aggacttttc gccaagggta gagatcctaa ggttgaaggt 7561 gaaagataca gaaatgctgg atgggaagcg taccaaaaga gaatcagcgc tccccattaa 7621 ttatacagga aacttaatag aacaaatcac atatttaatc taatagccac ctgcattggc 7681 acggtgcaac actacttcaa cttcatctta caaaaagatc acgtgatctg ttgtattggg 7741 atctctagac ctaataactt cgtatagcat acattatacg aagttatatt aagggttgct 7801 cgaccagaaa gcgaccaccc aactttggct gataatagcg tataaacaat gcatactttg 7861 tacgttcaaa atacaatgca gtagatatat ttatgcatat tacatataat acatatcaca 7921 taggaagcaa caggcgcgtt ggacttttaa ttttcgagga ccgcgaatcc ttacatcaca 7981 cccaatcccc cacaagtgat ccc //