LOCUS       sequence-150340-        7449 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector for stable expression of osTIR1-9Myc in mammalian 
            cell lines.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7449)
  AUTHORS   Lambrus BG, Moyer TC, Holland AJ
  TITLE     Applying the auxin-inducible degradation system for rapid protein 
            depletion in mammalian cells.
  JOURNAL   Methods Cell Biol. 2018;144:107-135. doi: 
            10.1016/bs.mcb.2018.03.004. Epub 2018 Apr 5.
  PUBMED    29804665
REFERENCE   2  (bases 1 to 7449)
  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"; journalName: "Methods 
            Cell Biol."; date: "2018"; volume: "144:107-135. doi"; pages: " 
            10.1016/bs.mcb.2018.03.004. Epub 2018 Apr 5"
COMMENT     Sequence Label: pBabe Neo osTIR1-9Myc
FEATURES             Location/Qualifiers
     source          1..7449
                     /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"
     CDS             1387..3111
                     /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             3139..3168
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3175..3204
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3211..3240
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3259..3288
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3295..3324
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3331..3360
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3379..3408
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3415..3444
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3451..3480
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3546..3566)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3551..3880
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3731..3866
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3890..4684
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3944..3963)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4554..4573
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             4730..5323
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5524..5541)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5695..6283)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5775..5794)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6461..7252,7253..7321))
                     /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     7084..7103
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7322..7426)
                     /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 gccaccatga cgtacttccc ggaggaggtg gtggagcaca tcttcagctt cctgccggcg
     1441 cagcgcgacc gcaacacggt ctcgctcgtc tgcaaggtgt ggtacgagat cgagaggctg
     1501 agccgccgcg gcgtcttcgt gggcaactgc tacgccgtgc gcgccggccg cgtcgccgcg
     1561 cggttcccca acgtgcgggc gctcacggtg aaggggaagc cccacttcgc cgacttcaac
     1621 ctcgtgcccc ccgactgggg cggctacgcg gggccgtgga tcgaggcggc cgcgagggga
     1681 tgccacggcc tggaggagct caggatgaag cggatggtgg tgtccgacga gagcctcgag
     1741 ctgctggctc gctcgttccc gcggttcagg gctcttgttc ttatcagctg cgaggggttc
     1801 agcactgacg ggctagccgc cgtcgcgagc cattgcaagc ttctgaggga gttggatttg
     1861 caggaaaatg aagtggagga tcgagggcct aggtggcttt cctgcttccc tgattcctgc
     1921 acatcacttg tctcattgaa ttttgcctgc atcaaagggg aggttaatgc tggttcactg
     1981 gagagacttg ttagcaggtc cccaaacctg cggagtttga ggctgaatcg atctgtatcg
     2041 gtagatacac ttgcaaagat actactgcgt acccctaact tggaggattt ggggacaggg
     2101 aatttgacag atgacttcca aactgagtcc tactttaagc ttaccagtgc tctggagaaa
     2161 tgcaagatgt tgaggagttt gtctggattc tgggatgctt ctcctgtttg cctgtcattt
     2221 atctaccccc tgtgtgctca actgacagga ttgaacttga gctatgcacc cacacttgat
     2281 gcttctgacc ttacaaaaat gattagccgc tgtgtgaagc tccaacgcct ttgggtactg
     2341 gattgtatct cggacaaagg cttgcaagtg gtggcctcca gttgcaaaga cttgcaagaa
     2401 ctcagggtat ttccatcaga tttctacgta gctggttatt ctgcagtgac agaggaggga
     2461 cttgttgcag tatccttggg ctgtccaaaa ctgaactcac tactgtactt ctgtcaccaa
     2521 atgactaatg ctgcactagt tactgtcgcc aagaactgtc caaatttcac acgattcaga
     2581 ctttgtattc ttgagccagg gaagcctgat gttgtgacaa gccaaccatt agatgaaggc
     2641 tttggagcta ttgttcgtga gtgcaaggga ttacaacgtt tgtcaatatc tggtcttctc
     2701 acagacaaag ttttcatgta tattgggaaa tatgcaaaac aacttgagat gctttctata
     2761 gcatttgctg gtgacagtga taagggtatg atgcatgtta tgaatggatg caagaattta
     2821 aggaaactgg agataagaga tagcccgttt ggtgatgctg cactcttggg gaattttgct
     2881 aggtacgaga caatgcgatc cctttggatg tcatcttgca atgtcacgtt aaaggggtgc
     2941 caagtccttg cgtcaaagat gccgatgctc aatgttgagg tcataaatga gcgggatggt
     3001 agcaatgaaa tggaggaaaa ccatggagat ctgcctaaag tggagaaatt atatgtgtac
     3061 cgcacaactg ctggggcgag ggatgatgca ccaaattttg ttaaaatcct agtcgactcc
     3121 ggttctgctg ctagtggtga acaaaagttg atttctgaag aagatttgaa cggtgaacaa
     3181 aagctaatct ccgaggaaga cttgaacggt gaacaaaaat taatctcaga agaagacttg
     3241 aacggatcct ctagaggtga acaaaagttg atttctgaag aagatttgaa cggtgaacaa
     3301 aagctaatct ccgaggaaga cttgaacggt gaacaaaaat taatctcaga agaagacttg
     3361 aacggatcct ctagaggtga acaaaagttg atttctgaag aagatttgaa cggtgaacaa
     3421 aagctaatct ccgaggaaga cttgaacggt gaacaaaaat taatctcaga agaagacttg
     3481 aacggatcca ctagctaaaa agtggaacga tcattcgaat tcgccagcac agtggtcgac
     3541 cctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3601 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3661 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3721 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3781 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     3841 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttacca tgattgaaca
     3901 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     3961 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4021 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     4081 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4141 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4201 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4261 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4321 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4381 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     4441 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4501 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4561 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4621 yggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4681 ctgaatcgat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
     4741 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaaata
     4801 cataactgag aatagagaag ttcagatcaa ggtcaggaac agatggaaca gctgaatatg
     4861 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
     4921 gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag
     4981 ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta gagaaccatc
     5041 agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca
     5101 atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc
     5161 cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg gtacccgtgt
     5221 atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc ttgggagggt
     5281 ctcctctgag tgattgacta cccgtcagcg ggggtctttc atttccgact tgtggtctcg
     5341 ctgccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt cttcacatgc
     5401 agcatgtatc aaaattaatt tggttttttt tcttaagtat ttacattaaa tggccatagt
     5461 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggc gctcttccgc
     5521 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5581 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5641 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5701 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5761 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5821 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5881 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5941 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     6001 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     6061 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     6121 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     6181 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6241 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6301 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6361 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt tgcggccgca
     6421 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6481 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6541 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6601 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6661 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6721 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6781 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6841 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6901 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6961 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7021 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac
     7081 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7141 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7201 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7261 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7321 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7381 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7441 aagtgccac
//
