LOCUS       sequence-362006-        4847 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  in vitro transcription of osTIR1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4847)
  AUTHORS   Camlin NJ, Evans JP
  TITLE     Auxin-inducible protein degradation as a novel approach for protein 
            depletion and reverse genetic discoveries in mammalian 
            oocytesdagger.
  JOURNAL   Biol Reprod. 2019 Oct 25;101(4):704-718. doi: 10.1093/biolre/ioz113.
  PUBMED    31299080
REFERENCE   2  (bases 1 to 4847)
  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"; journalName: 
            "Biol Reprod"; date: "2019-10-25- 25"; volume: "101"; issue: "4"; 
            pages: "704-718"
COMMENT     Sequence Label: pIVT-osTIR1-9myc
FEATURES             Location/Qualifiers
     source          1..4847
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     3'UTR           complement(32..157)
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     CDS             complement(246..275)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(282..311)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(318..347)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(375..2099)
                     /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"
     5'UTR           complement(2130..2172)
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     primer_bind     complement(2214..2233)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2215..2233)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2245..2263)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2246..2263)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2287..2303)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2287..2303)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2300..2322)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2311..2327
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2335..2341,2342..2359,2360..2365))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2380..2401
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2518..2535)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2689..3277)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2769..2788)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3448..4239,4240..4308))
                     /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     4071..4090
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4309..4413)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4481..4499
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4537..4559)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4659..4678
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tttttttttt tttttttttt ttttttagaa tgtgaagaaa ctttcttttt attaggagca
       61 gatacgaatg gctacatttt gggggacaac attttgtaaa gtgtaagttg gtattatgta
      121 gcttagagac tccattcggg tgttcttgag gctggtttag tggtaaccag atcctagtca
      181 gtcactagaa gggcgaattc gagctcggta cccggggatc ctctagatta gctagtggat
      241 ccgttcaagt cttcttctga gattaatttt tgttcaccgt tcaagtcttc ctcggagatt
      301 agcttttgtt caccgttcaa atcttcttca gaaatcaact tttgttcacc actagcagca
      361 gaaccggagt cgactaggat tttaacaaaa tttggtgcat catccctcgc cccagcagtt
      421 gtgcggtaca catataattt ctccacttta ggcagatctc catggttttc ctccatttca
      481 ttgctaccat cccgctcatt tatgacctca acattgagca tcggcatctt tgacgcaagg
      541 acttggcacc cctttaacgt gacattgcaa gatgacatcc aaagggatcg cattgtctcg
      601 tacctagcaa aattccccaa gagtgcagca tcaccaaacg ggctatctct tatctccagt
      661 ttccttaaat tcttgcatcc attcataaca tgcatcatac ccttatcact gtcaccagca
      721 aatgctatag aaagcatctc aagttgtttt gcatatttcc caatatacat gaaaactttg
      781 tctgtgagaa gaccagatat tgacaaacgt tgtaatccct tgcactcacg aacaatagct
      841 ccaaagcctt catctaatgg ttggcttgtc acaacatcag gcttccctgg ctcaagaata
      901 caaagtctga atcgtgtgaa atttggacag ttcttggcga cagtaactag tgcagcatta
      961 gtcatttggt gacagaagta cagtagtgag ttcagttttg gacagcccaa ggatactgca
     1021 acaagtccct cctctgtcac tgcagaataa ccagctacgt agaaatctga tggaaatacc
     1081 ctgagttctt gcaagtcttt gcaactggag gccaccactt gcaagccttt gtccgagata
     1141 caatccagta cccaaaggcg ttggagcttc acacagcggc taatcatttt tgtaaggtca
     1201 gaagcatcaa gtgtgggtgc atagctcaag ttcaatcctg tcagttgagc acacaggggg
     1261 tagataaatg acaggcaaac aggagaagca tcccagaatc cagacaaact cctcaacatc
     1321 ttgcatttct ccagagcact ggtaagctta aagtaggact cagtttggaa gtcatctgtc
     1381 aaattccctg tccccaaatc ctccaagtta ggggtacgca gtagtatctt tgcaagtgta
     1441 tctaccgata cagatcgatt cagcctcaaa ctccgcaggt ttggggacct gctaacaagt
     1501 ctctccagtg aaccagcatt aacctcccct ttgatgcagg caaaattcaa tgagacaagt
     1561 gatgtgcagg aatcagggaa gcaggaaagc cacctaggcc ctcgatcctc cacttcattt
     1621 tcctgcaaat ccaactccct cagaagcttg caatggctcg cgacggcggc tagcccgtca
     1681 gtgctgaacc cctcgcagct gataagaaca agagccctga accgcgggaa cgagcgagcc
     1741 agcagctcga ggctctcgtc ggacaccacc atccgcttca tcctgagctc ctccaggccg
     1801 tggcatcccc tcgcggccgc ctcgatccac ggccccgcgt agccgcccca gtcggggggc
     1861 acgaggttga agtcggcgaa gtggggcttc cccttcaccg tgagcgcccg cacgttgggg
     1921 aaccgcgcgg cgacgcggcc ggcgcgcacg gcgtagcagt tgcccacgaa gacgccgcgg
     1981 cggctcagcc tctcgatctc gtaccacacc ttgcagacga gcgagaccgt gttgcggtcg
     2041 cgctgcgccg gcaggaagct gaagatgtgc tccaccacct cctccgggaa gtacgtcatg
     2101 tcgacctgca ggcacgcaag cttgatctgc caaagttgag cgtttattct gagcttctgc
     2161 aaaaagaaca agcaagcttc tcctcccttt agtgagggta attctctctc tctccctata
     2221 gtgagtcgta ttaattcctt ctcttctata gtgtcaccta aatcgttgga gcttggcgta
     2281 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     2341 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     2401 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     2461 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     2521 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2581 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2641 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2701 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2761 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2821 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2881 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2941 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     3001 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     3061 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3121 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3181 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3241 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3301 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3361 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3421 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3481 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3541 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     3601 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3661 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3721 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3781 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3841 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3901 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3961 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4021 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4081 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4141 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4201 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4261 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4321 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4381 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     4441 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     4501 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     4561 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     4621 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     4681 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     4741 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     4801 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgat
//
