LOCUS       sequence-409726-        5407 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains OsTiR1 ORF and LoxP and Lox2272 sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5407)
  AUTHORS   Kainov Y, Zhuravskaya A, Makeyev EV
  TITLE     Protocol for auxin-inducible depletion of the RNA-binding protein 
            PTBP1 in mouse embryonic stem cells.
  JOURNAL   STAR Protoc. 2023 Oct 18;4(4):102644. doi: 
            10.1016/j.xpro.2023.102644.
  PUBMED    37862173
REFERENCE   2  (bases 1 to 5407)
  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: "STAR 
            Protoc."; date: "2023-10-18"; volume: "4(4):102644. doi"; pages: " 
            10.1016/j.xpro.2023.102644"
COMMENT     Sequence Label: pML33
FEATURES             Location/Qualifiers
     source          1..5407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        19..518
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(54..77)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     451..470
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      553..562
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(568..601)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(604..637)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     639..655
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     640..656
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             681..2408
                     /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"
     primer_bind     complement(2479..2495)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2479..2495)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     polyA_signal    2519..2640
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2530..2549)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2584..2603
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2823..3278
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2910..2929)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3120..3141
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3345..3364)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3464..3486
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3524..3542)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3610..3714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3715..3783,3784..4575)
                     /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     complement(3933..3952)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4746..5334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5235..5254
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgacctcgaa attctaccgg gtaggggagg cgcttttccc aaggcagtct ggagcatgcg
       61 ctttagcagc cccgctgggc acttggcgct acacaagtgg cctctggcct cgcacacatt
      121 ccacatccac cggtaggcgc caaccggctc cgttctttgg tggccccttc gcgccacctt
      181 ctactcctcc cctagtcagg aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt
      241 gacaaatgga agtagcacgt ctcactagtc tcgtgcagat ggacagcacc gctgagcaat
      301 ggaagcgggt aggcctttgg ggcagcggcc aatagcagct ttgctccttc gctttctggg
      361 ctcagaggct gggaaggggt gggtccgggg gcgggctcag gggcgggctc aggggcgggg
      421 cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg
      481 ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac ctgcatccat ctagatctcg
      541 aaggatctgg aggccaccat ggtgtcgata acttcgtata gcatacatta tacgaagtta
      601 tcgataactt cgtataggat actttatacg aagttatctc gaggtcgacg gtatcgataa
      661 gcttgattag cgctgccacc atgacatact ttcctgaaga ggtcgtcgaa cacattttta
      721 gcttcctgcc tgcacagaga gatagaaaca cagtgagcct ggtctgcaaa gtgtggtacg
      781 agatcgaacg cctgagccgg agaggagtgt tcgtcggcaa ctgctatgct gtgagagcag
      841 gcagggtcgc cgctaggttt ccaaatgtgc gcgcactgac cgtcaagggg aaaccccact
      901 tcgccgactt taacctggtg ccccctgatt ggggaggata cgccggccct tggatcgagg
      961 cagccgctcg cggctgtcat ggactggagg aactgcgcat gaagcgaatg gtggtctctg
     1021 acgaaagtct ggagctgctg gctcggagct tccctaggtt tcgcgcactg gtgctgattt
     1081 cttgcgaagg cttcagcacc gatggactgg cagccgtggc ctcccactgt aagctgctgc
     1141 gggagctgga cctccaggag aatgaagtgg aggatagagg ccccagatgg ctgtcttgct
     1201 tcccagactc atgtaccagc ctggtgtccc tgaactttgc ctgcatcaaa ggcgaagtga
     1261 atgctgggtc cctggagcgg ctggtctcaa gaagccccaa cctgaggtct ctgcggctga
     1321 accggagcgt gagcgtggac actctggcta agattctgct gagaacccct aacctggagg
     1381 atctgggaac cggcaatctg acagacgatt tccagacaga atcctacttt aaactgactt
     1441 ctgccctgga gaagtgtaaa atgctgagga gtctgtcagg attctgggat gcttcacccg
     1501 tgtgcctgag ctttatctac cctctgtgtg cacagctgac aggcctgaac ctgagctatg
     1561 caccaaccct ggacgccagt gatctgacaa agatgatctc acgctgcgtg aaactccagc
     1621 gactgtgggt gctggactgt atttccgata aggggctcca ggtggtcgcc agctcctgca
     1681 aggacctcca ggagctgaga gtgttcccat ctgattttta cgtggccgga tatagtgctg
     1741 tcactgagga aggcctggtg gcagtctcac tgggatgccc aaagctgaac agcctgctgt
     1801 atttctgtca tcagatgact aatgctgcac tggtgaccgt cgccaagaac tgccctaatt
     1861 tcacccgatt tcggctgtgt attctggaac caggcaaacc cgacgtggtc acatcccagc
     1921 cactggatga agggtttgga gctatcgtga gagagtgcaa gggactccag aggctgagca
     1981 tttccggcct gctgacagac aaagtgttca tgtacatcgg caagtatgct aagcagctgg
     2041 agatgctgag cattgcattt gccggagact ccgataaggg catgatgcac gtgatgaacg
     2101 ggtgtaagaa tctgcgaaaa ctggaaatcc gggacagccc tttcggggat gccgctctgc
     2161 tgggaaactt tgccagatac gagacaatga ggagcctgtg gatgtctagt tgcaatgtga
     2221 ctctgaaggg ctgtcaggtc ctggctagta aaatgcctat gctgaacgtg gaagtcatta
     2281 atgagcggga cgggtctaac gaaatggagg aaaatcatgg cgacctgcca aaggtggaga
     2341 aactgtatgt gtatcggacc accgcagggg caagagatga tgctcccaac tttgtgaaga
     2401 ttctgtgagg atccatcgga tcccgggccc gtcgacggta caagtaatga attaattaag
     2461 aattcctgca gcccggggga tccactagtt ctagagcggc cgcttcgagc agacatgata
     2521 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     2581 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     2641 aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt
     2701 taaagcaagt aaaacctcta caaatgtggt aaaatcgcga taaggatccg ggctggcgta
     2761 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     2821 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2881 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2941 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3001 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3061 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3121 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3181 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3241 taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt
     3301 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat
     3361 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3421 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3481 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     3541 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     3601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     3721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     3781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     3961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     4021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     4081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     4141 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     4201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     4261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     4321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     4381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     4441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     4501 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     4561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     4621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4861 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     4921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     5041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     5101 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     5161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     5221 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     5281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catggctcga
     5401 cagatct
//
