LOCUS       sequence-108204-        7352 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For retroviral expression of E3 ubiquitin ligase osTIR1 with N-term 
            NES and C-term 9x myc tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7352)
  AUTHORS   Holland AJ, Fachinetti D, Han JS, Cleveland DW
  TITLE     Inducible, reversible system for the rapid and complete degradation 
            of proteins in mammalian cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2012 Dec 4;109(49):E3350-7. doi: 
            10.1073/pnas.1216880109. Epub 2012 Nov 12.
  PUBMED    23150568
REFERENCE   2  (bases 1 to 7352)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1073/pnas.1216880109"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2012-12-4- 4"; volume: "109"; issue: "49"; pages: "E3350-7"
COMMENT     Sequence Label: pBabe TIR1-9myc
FEATURES             Location/Qualifiers
     source          1..7352
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             934..1350
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1318..1334
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1381..1390
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1435..3156
                     /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             3184..3213
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3220..3249
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3256..3285
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3304..3333
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3340..3369
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3376..3405
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3424..3453
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3460..3489
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3496..3525
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3591..3611)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3596..3925
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3776..3911
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3935..4534
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(3935..3954)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4431..4451
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4633..5226
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5427..5444)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5598..6186)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5678..5697)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6364..7155,7156..7224))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6987..7006
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7225..7329)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 taagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
      721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
      781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
      841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt
      901 ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt
      961 ttgaccttag atcactggaa agatgtcgag cggctcgctc acaaccagtc ggtagatgtc
     1021 aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg
     1081 ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca
     1141 cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct
     1201 tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttctt
     1261 ccatccgcgc cgtctctccc ccttgaacct cctctttcga ccccgcctca atcctccctt
     1321 tatccagccc tcactccttc tctaggcgcc ggccggatcc cagtgtggtg gtacatcgcc
     1381 gccaccatgg cattacaaaa aaaattagaa gaattagaat tagatgaagg cggcacgtac
     1441 ttcccggagg aggtggtgga gcacatcttc agcttcctgc cggcgcagcg cgaccgcaac
     1501 acggtctcgc tcgtctgcaa ggtgtggtac gagatcgaga ggctgagccg ccgcggcgtc
     1561 ttcgtgggca actgctacgc cgtgcgcgcc ggccgcgtcg ccgcgcggtt ccccaacgtg
     1621 cgggcgctca cggtgaaggg gaagccccac ttcgccgact tcaacctcgt gccccccgac
     1681 tggggcggct acgcggggcc gtggatcgag gcggccgcga ggggatgcca cggcctggag
     1741 gagctcagga tgaagcggat ggtggtgtcc gacgagagcc tcgagctgct ggctcgctcg
     1801 ttcccgcggt tcagggctct tgttcttatc agctgcgagg ggttcagcac tgacgggcta
     1861 gccgccgtcg cgagccattg caagcttctg agggagttgg atttgcagga aaatgaagtg
     1921 gaggatcgag ggcctaggtg gctttcctgc ttccctgatt cctgcacatc acttgtctca
     1981 ttgaattttg cctgcatcaa aggggaggtt aatgctggtt cactggagag acttgttagc
     2041 aggtccccaa acctgcggag tttgaggctg aatcgatctg tatcggtaga tacacttgca
     2101 aagatactac tgcgtacccc taacttggag gatttgggga cagggaattt gacagatgac
     2161 ttccaaactg agtcctactt taagcttacc agtgctctgg agaaatgcaa gatgttgagg
     2221 agtttgtctg gattctggga tgcttctcct gtttgcctgt catttatcta ccccctgtgt
     2281 gctcaactga caggattgaa cttgagctat gcccccacac ttgatgcttc tgaccttaca
     2341 aaaatgatta gccgctgtgt gaagctccaa cgcctttggg tactggattg tatctcggac
     2401 aaaggcttgc aagtggtggc ctccagttgc aaagacttgc aagaactcag ggtatttcca
     2461 tcagatttct acgtagctgg ttattctgca gtgacagagg agggacttgt tgcagtatcc
     2521 ttgggctgtc caaaactgaa ctcactactg tacttctgtc accaaatgac taatgctgca
     2581 ctagttactg tcgccaagaa ctgtccaaat ttcacacgat tcagactttg tattcttgag
     2641 ccagggaagc ctgatgttgt gacaagccaa ccattagatg aaggctttgg agctattgtt
     2701 cgtgagtgca agggattaca acgtttgtca atatctggtc ttctcacaga caaagttttc
     2761 atgtatattg ggaaatatgc aaaacaactt gagatgcttt ctatagcatt tgctggtgac
     2821 agtgataagg gtatgatgca tgttatgaat ggatgcaaga atttaaggaa actggagata
     2881 agagatagcc cgtttggtga tgctgcactc ttggggaatt ttgctaggta cgagacaatg
     2941 cgatcccttt ggatgtcatc ttgcaatgtc acgttaaagg ggtgccaagt ccttgcgtca
     3001 aagatgccga tgctcaatgt tgaggtcata aatgagcggg atggtagcaa tgaaatggag
     3061 gaaaaccatg gagatctgcc taaagtggag aaattatatg tgtaccgcac aactgctggg
     3121 gcgagggatg atgcaccaaa ttttgttaaa atcctagtcg actccggttc tgctgctagt
     3181 ggtgaacaaa agttgatttc tgaagaagat ttgaacggtg aacaaaagct aatctccgag
     3241 gaagacttga acggtgaaca aaaattaatc tcagaagaag acttgaacgg atcctctaga
     3301 ggtgaacaaa agttgatttc tgaagaagat ttgaacggtg aacaaaagct aatctccgag
     3361 gaagacttga acggtgaaca aaaattaatc tcagaagaag acttgaacgg atcctctaga
     3421 ggtgaacaaa agttgatttc tgaagaagat ttgaacggtg aacaaaagct aatctccgag
     3481 gaagacttga acggtgaaca aaaattaatc tcagaagaag acttgaacgg atccactagc
     3541 taaaaagtgg aacgatcatt cgaattcgcc agcacagtgg tcgaccctgt ggaatgtgtg
     3601 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3661 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3721 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3781 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3841 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     3901 ttttggaggc ctaggctttt gcaaaaagct taccatgacc gagtacaagc ccacggtgcg
     3961 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     4021 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     4081 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     4141 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     4201 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     4261 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     4321 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     4381 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     4441 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gagtgcccga aggaccgcgc
     4501 gacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc
     4561 gaccgaaagg agcgcacgac cccatgcatc gataaaataa aagattttat ttagtctcca
     4621 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     4681 attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg
     4741 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     4801 cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt
     4861 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc
     4921 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc
     4981 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc
     5041 tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga
     5101 ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg
     5161 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct
     5221 ttcatttccg acttgtggtc tcgctgcctt gggagggtct cctctgagtg attgactacc
     5281 cgtcagcggg ggtcttcaca tgcagcatgt atcaaaatta atttggtttt ttttcttaag
     5341 tatttacatt aaatggccat agttgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5401 tttgcgtatt ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5461 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5521 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5581 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5641 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5701 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5761 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5821 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5881 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5941 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6001 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     6061 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6121 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6181 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6241 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6301 taaaaatgaa gtttgcggcc gcaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6361 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6421 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6481 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6541 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6601 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6661 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6721 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6781 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6841 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6901 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6961 ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa ctttaaaagt
     7021 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7081 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7141 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7201 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7261 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7321 ggttccgcgc acatttcccc gaaaagtgcc ac
//
