LOCUS       sequence-182589-        7320 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For retroviral expression of E3 ubiquitin ligase osTIR1 with 9x myc 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7320)
  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 7320)
  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..7320
                     /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             942..1358
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1288..1310
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1326..1342
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1395..3119
                     /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"
     CDS             3147..3176
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3183..3212
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3219..3248
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3267..3296
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3303..3332
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3339..3368
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3387..3416
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3423..3452
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3459..3488
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3554..3574)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3559..3888
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3739..3874
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3898..4497
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3898..3917)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4394..4414
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5394..5411)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5565..6153)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5645..5664)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6332..7123,7124..7192))
                     /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     6955..6974
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7193..7297)
                     /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 caagctggcc 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 tgctgcagcg ctgcagcatc gttctgtgtt
      901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
      961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatccca gtgtggtggt
     1381 acatcgccgc caccatgacg tacttcccgg aggaggtggt ggagcacatc ttcagcttcc
     1441 tgccggcgca gcgcgaccgc aacacggtct cgctcgtctg caaggtgtgg tacgagatcg
     1501 agaggctgag ccgccgcggc gtcttcgtgg gcaactgcta cgccgtgcgc gccggccgcg
     1561 tcgccgcgcg gttccccaac gtgcgggcgc tcacggtgaa ggggaagccc cacttcgccg
     1621 acttcaacct cgtgcccccc gactggggcg gctacgcggg gccgtggatc gaggcggccg
     1681 cgaggggatg ccacggcctg gaggagctca ggatgaagcg gatggtggtg tccgacgaga
     1741 gcctcgagct gctggctcgc tcgttcccgc ggttcagggc tcttgttctt atcagctgcg
     1801 aggggttcag cactgacggg ctagccgccg tcgcgagcca ttgcaagctt ctgagggagt
     1861 tggatttgca ggaaaatgaa gtggaggatc gagggcctag gtggctttcc tgcttccctg
     1921 attcctgcac atcacttgtc tcattgaatt ttgcctgcat caaaggggag gttaatgctg
     1981 gttcactgga gagacttgtt agcaggtccc caaacctgcg gagtttgagg ctgaatcgat
     2041 ctgtatcggt agatacactt gcaaagatac tactgcgtac ccctaacttg gaggatttgg
     2101 ggacagggaa tttgacagat gacttccaaa ctgagtccta ctttaagctt accagtgctc
     2161 tggagaaatg caagatgttg aggagtttgt ctggattctg ggatgcttct cctgtttgcc
     2221 tgtcatttat ctaccccctg tgtgctcaac tgacaggatt gaacttgagc tatgcaccca
     2281 cacttgatgc ttctgacctt acaaaaatga ttagccgctg tgtgaagctc caacgccttt
     2341 gggtactgga ttgtatctcg gacaaaggct tgcaagtggt ggcctccagt tgcaaagact
     2401 tgcaagaact cagggtattt ccatcagatt tctacgtagc tggttattct gcagtgacag
     2461 aggagggact tgttgcagta tccttgggct gtccaaaact gaactcacta ctgtacttct
     2521 gtcaccaaat gactaatgct gcactagtta ctgtcgccaa gaactgtcca aatttcacac
     2581 gattcagact ttgtattctt gagccaggga agcctgatgt tgtgacaagc caaccattag
     2641 atgaaggctt tggagctatt gttcgtgagt gcaagggatt acaacgtttg tcaatatctg
     2701 gtcttctcac agacaaagtt ttcatgtata ttgggaaata tgcaaaacaa cttgagatgc
     2761 tttctatagc atttgctggt gacagtgata agggtatgat gcatgttatg aatggatgca
     2821 agaatttaag gaaactggag ataagagata gcccgtttgg tgatgctgca ctcttgggga
     2881 attttgctag gtacgagaca atgcgatccc tttggatgtc atcttgcaat gtcacgttaa
     2941 aggggtgcca agtccttgcg tcaaagatgc cgatgctcaa tgttgaggtc ataaatgagc
     3001 gggatggtag caatgaaatg gaggaaaacc atggagatct gcctaaagtg gagaaattat
     3061 atgtgtaccg cacaactgct ggggcgaggg atgatgcacc aaattttgtt aaaatcctag
     3121 tcgactccgg ttctgctgct agtggtgaac aaaagttgat ttctgaagaa gatttgaacg
     3181 gtgaacaaaa gctaatctcc gaggaagact tgaacggtga acaaaaatta atctcagaag
     3241 aagacttgaa cggatcctct agaggtgaac aaaagttgat ttctgaagaa gatttgaacg
     3301 gtgaacaaaa gctaatctcc gaggaagact tgaacggtga acaaaaatta atctcagaag
     3361 aagacttgaa cggatcctct agaggtgaac aaaagttgat ttctgaagaa gatttgaacg
     3421 gtgaacaaaa gctaatctcc gaggaagact tgaacggtga acaaaaatta atctcagaag
     3481 aagacttgaa cggatccact agctaaaaag tggaacgatc attcgaattc gccagcacag
     3541 tggtcgaccc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3601 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3661 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3721 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3781 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3841 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttaccatg
     3901 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     3961 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     4021 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     4081 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     4141 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     4201 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     4261 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     4321 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     4381 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     4441 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc
     4501 ccgccccacg acccgcagcg cccgaccgaa aggagcgcac gaccccatgc atcgataaaa
     4561 taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt
     4621 ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata
     4681 gagaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc aaacaggata
     4741 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat
     4801 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     4861 ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg
     4921 tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct
     4981 cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact
     5041 cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct
     5101 cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
     5161 tgactacccg tcagcggggg tctttcgcat ccgacttgtg gtctcgctgt tccttgggag
     5221 ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcacatgca gcatgtatca
     5281 aaattaattt ggtttttttt cttaagtatt tacattaaat ggccatagtt gcattaatga
     5341 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5401 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5461 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5521 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5581 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5641 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5701 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5761 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5821 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5881 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5941 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6001 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6061 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6121 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6181 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6241 aggatcttca cctagatcct tttgcggccg caaatcaatc taaagtatat atgagaatct
     6301 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6361 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6421 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6481 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6541 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6601 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6661 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6721 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6781 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6841 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6901 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6961 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7021 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7081 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7141 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7201 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7261 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
//
