LOCUS Exported 4824 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pMVP R4-R3 entry plasmid, contains osTir1 for 3- or 4-component MultiSite Gateway Pro assembly. ACCESSION . VERSION . KEYWORDS KJ101: pMVP (R4-R3) osTir1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4824) AUTHORS Haldeman JM, Conway AE, Arlotto ME, Slentz DH, Muoio DM, Becker TC, Newgard CB TITLE Creation of versatile cloning platforms for transgene expression and dCas9-based epigenome editing. JOURNAL Nucleic Acids Res. 2018 Dec 27. pii: 5264291. doi: 10.1093/nar/gky1286. PUBMED 30590691 REFERENCE 2 (bases 1 to 4824) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4824 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 92..109 /label=L4440 /note="L4440 vector, forward primer" terminator 268..295 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 387..473 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 522..544 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 536..553 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 537..553 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind complement(603..727) /gene="mutant version of attR" /label=attR4 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 777..2498 /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="TYFPEEVVEHIFSFLPAQRDRNTVSLVCKVWYEIERLSRRGVFVG NCYAVRAGRVAARFPNVRALTVKGKPHFADFNLVPPDWGGYAGPWIEAAARGCHGLEEL RMKRMVVSDESLELLARSFPRFRALVLISCEGFSTDGLAAVASHCKLLRELDLQENEVE DRGPRWLSCFPDSCTSLVSLNFACIKGEVNAGSLERLVSRSPNLRSLRLNRSVSVDTLA KILLRTPNLEDLGTGNLTDDFQTESYFKLTSALEKCKMLRSLSGFWDASPVCLSFIYPL CAQLTGLNLSYAPTLDASDLTKMISRCVKLQRLWVLDCISDKGLQVVASSCKDLQELRV FPSDFYVAGYSAVTEEGLVAVSLGCPKLNSLLYFCHQMTNAALVTVAKNCPNFTRFRLC ILEPGKPDVVTSQPLDEGFGAIVRECKGLQRLSISGLLTDKVFMYIGKYAKQLEMLSIA FAGDSDKGMMHVMNGCKNLRKLEIRDSPFGDAALLGNFARYETMRSLWMSSCNVTLKGC QVLASKMPMLNVEVINERDGSNEMEENHGDLPKVEKLYVYRTTAGARDDAPNFVKIL" CDS 2526..2555 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 2562..2591 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 2598..2627 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 2646..2675 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 2682..2711 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 2718..2747 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 2766..2795 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 2802..2831 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 2838..2867 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" protein_bind 2887..3010 /gene="mutant version of attR" /label=attR3 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(3065..3084) /label=T7 /note="T7 promoter, forward primer" promoter complement(3066..3084) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3089..3105) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3089..3105) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3207..3226) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 3218..4027 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(3285..3304) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 4174..4762 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4663..4682 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga 61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga 121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc 241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta 301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc 361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa 421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg 481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa 541 aacgacggcc agtcttaagc tcgggcccct acaggtcact aataccatct aagtagttga 601 ttcatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa 661 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcaact tttctataca 721 aagttgccat ggcattacaa aaaaaattag aagaattaga attagatgaa ggcggcacgt 781 acttcccgga ggaggtggtg gagcacatct tcagcttcct gccggcgcag cgcgaccgca 841 acacggtctc gctcgtctgc aaggtgtggt acgagatcga gaggctgagc cgccgcggcg 901 tcttcgtggg caactgctac gccgtgcgcg ccggccgcgt cgccgcgcgg ttccccaacg 961 tgcgggcgct cacggtgaag gggaagcccc acttcgccga cttcaacctc gtgccccccg 1021 actggggcgg ctacgcgggg ccgtggatcg aggcggccgc gaggggatgc cacggcctgg 1081 aggagctcag gatgaagcgg atggtggtgt ccgacgagag cctcgagctg ctggctcgct 1141 cgttcccgcg gttcagggct cttgttctta tcagctgcga ggggttcagc actgacgggc 1201 tagccgccgt cgcgagccat tgcaagcttc tgagggagtt ggatttgcag gaaaatgaag 1261 tggaggatcg agggcctagg tggctttcct gcttccctga ttcctgcaca tcacttgtct 1321 cattgaattt tgcctgcatc aaaggggagg ttaatgctgg ttcactggag agacttgtta 1381 gcaggtcccc aaacctgcgg agtttgaggc tgaatcgatc tgtatcggta gatacacttg 1441 caaagatact actgcgtacc cctaacttgg aggatttggg gacagggaat ttgacagatg 1501 acttccaaac tgagtcctac tttaagctta ccagtgctct ggagaaatgc aagatgttga 1561 ggagtttgtc tggattctgg gatgcttctc ctgtttgcct gtcatttatc taccccctgt 1621 gtgctcaact gacaggattg aacttgagct atgcacccac acttgatgct tctgacctta 1681 caaaaatgat tagccgctgt gtgaagctcc aacgcctttg ggtactggat tgtatctcgg 1741 acaaaggctt gcaagtggtg gcctccagtt gcaaagactt gcaagaactc agggtatttc 1801 catcagattt ctacgtagct ggttattctg cagtgacaga ggagggactt gttgcagtat 1861 ccttgggctg tccaaaactg aactcactac tgtacttctg tcaccaaatg actaatgctg 1921 cactagttac tgtcgccaag aactgtccaa atttcacacg attcagactt tgtattcttg 1981 agccagggaa gcctgatgtt gtgacaagcc aaccattaga tgaaggcttt ggagctattg 2041 ttcgtgagtg caagggatta caacgtttgt caatatctgg tcttctcaca gacaaagttt 2101 tcatgtatat cgggaaatat gcaaaacaac ttgagatgct ttctatagca tttgctggtg 2161 acagtgataa gggtatgatg catgttatga atggatgcaa gaatttaagg aaactggaga 2221 taagagatag cccgtttggt gatgctgcac tcttggggaa ttttgctagg tacgagacaa 2281 tgcgatccct ttggatgtca tcttgcaatg tcacgttaaa ggggtgccaa gtccttgcgt 2341 caaagatgcc gatgctcaat gttgaggtca taaatgagcg ggatggtagc aatgaaatgg 2401 aggaaaacca tggagatctg cctaaagtgg agaaattata tgtgtaccgc acaactgctg 2461 gggcgaggga tgatgcacca aattttgtta aaatcctagt cgactccggt tctgctgcta 2521 gtggtgaaca aaagttgatt tctgaagaag atttgaacgg tgaacaaaag ctaatctccg 2581 aggaagactt gaacggtgaa caaaaattaa tctcagaaga agacttgaac ggatcctcta 2641 gaggtgaaca aaagttgatt tctgaagaag atttgaacgg tgaacaaaag ctaatctccg 2701 aggaagactt gaacggtgaa caaaaattaa tctcagaaga agacttgaac ggatcctcta 2761 gaggtgaaca aaagttgatt tctgaagaag atttgaacgg tgaacaaaag ctaatctccg 2821 aggaagactt gaacggtgaa caaaaattaa tctcagaaga agacttgaac ggatccacta 2881 gctaaacaac tttgtataat aaagttgaac gagaaacgta aaatgatata aatatcaata 2941 tattaaatta gattttgcat aaaaaacaga ctacataata ctgtaaaaca caacatatcc 3001 agtcactatg aatcaactac ttagatggta ttagtgacct gtactgcagg gcggccgcga 3061 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt 3121 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa 3181 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac 3241 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg 3301 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc 3361 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt 3421 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac 3481 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt 3541 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg 3601 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc 3661 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg 3721 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc 3781 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa 3841 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc 3901 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa 3961 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt 4021 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg 4081 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc 4141 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat 4201 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga 4261 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt 4321 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 4381 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 4441 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 4501 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 4561 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 4621 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct 4681 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc 4741 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt 4801 ttgctggcct tttgctcaca tgtt //