LOCUS       Exported                4821 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  pMVP R4-R3 entry plasmid, contains osTir1 (no stop codon) for 3- or 
            4-component MultiSite Gateway Pro assembly.
ACCESSION   .
VERSION     .
KEYWORDS    KJ201: pMVP (R4-R3) osTir1 open
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4821)
  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 4821)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4821
                     /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    2884..3007
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3062..3081)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3063..3081)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3086..3102)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3086..3102)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3204..3223)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3215..4024
                     /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(3282..3301)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4171..4759
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4660..4679
                     /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 gcacaacttt gtataataaa gttgaacgag aaacgtaaaa tgatataaat atcaatatat
     2941 taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa catatccagt
     3001 cactatgaat caactactta gatggtatta gtgacctgta ctgcagggcg gccgcgatat
     3061 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     3121 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     3181 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     3241 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     3301 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     3361 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     3421 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     3481 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     3541 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     3601 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     3661 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     3721 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     3781 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     3841 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     3901 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     3961 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     4021 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     4081 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     4141 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4201 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4261 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     4321 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4381 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4441 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4501 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     4561 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     4621 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4681 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4741 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4801 ctggcctttt gctcacatgt t
//
