LOCUS       sequence-274198-        5021 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  in vitro transcription for mRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5021)
  AUTHORS   Borsos M, Perricone SM, Schauer T, Pontabry J, de Luca KL, de Vries 
            SS, Ruiz-Morales ER, Torres-Padilla ME, Kind J
  TITLE     Genome-lamina interactions are established de novo in the early 
            mouse embryo.
  JOURNAL   Nature. 2019 May;569(7758):729-733. doi: 10.1038/s41586-019-1233-0. 
            Epub 2019 May 22.
  PUBMED    31118510
REFERENCE   2  (bases 1 to 5021)
  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: 
            "10.1038/s41586-019-1233-0"; journalName: "Nature"; date: "2019-05";
            volume: "569"; issue: "7758"; pages: "729-733"
COMMENT     Sequence Label: pRN3P_TIR1
FEATURES             Location/Qualifiers
     source          1..5021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..20
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     5'UTR           45..87
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             102..1826
                     /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             1854..1883
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1890..1919
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1926..1955
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     3'UTR           1997..2122
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(2210..2229)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2211..2229)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2236..2253)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2236..2252)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2245..2267)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(2394..2849)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2531..2552)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2743..2762
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2876..2980
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2981..3049,3050..3841)
                     /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(3199..3218)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4012..4600
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4501..4520
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4754..4771
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4888..4909
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4924..4929,4930..4947,4948..4954)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4962..4978
                     /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)."
     primer_bind     4967..4989
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4986..5002
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4986..5002
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aaattaaccc tcactaaagg gaacaaaagc tggagctaag cttgcttgtt ctttttgcag
       61 aagctcagaa taaacgctca actttggcag atctggccgc catgacgtac ttcccggagg
      121 aggtggtgga gcacatcttc agcttcctgc cggcgcagcg cgaccgcaac acggtctcgc
      181 tcgtctgcaa ggtgtggtac gagatcgaga ggctgagccg ccgcggcgtc ttcgtgggca
      241 actgctacgc cgtgcgcgcc ggccgcgtcg ccgcgcggtt ccccaacgtg cgggcgctca
      301 cggtgaaggg gaagccccac ttcgccgact tcaacctcgt gccccccgac tggggcggct
      361 acgcggggcc gtggatcgag gcggccgcga ggggatgcca cggcctggag gagctcagga
      421 tgaagcggat ggtggtgtcc gacgagagcc tcgagctgct ggctcgctcg ttcccgcggt
      481 tcagggctct tgttcttatc agctgcgagg ggttcagcac tgacgggcta gccgccgtcg
      541 cgagccattg caagcttctg agggagttgg atttgcagga aaatgaagtg gaggatcgag
      601 ggcctaggtg gctttcctgc ttccctgatt cctgcacatc acttgtctca ttgaattttg
      661 cctgcatcaa aggggaggtt aatgctggtt cactggagag acttgttagc aggtccccaa
      721 acctgcggag tttgaggctg aatcgatctg tatcggtaga tacacttgca aagatactac
      781 tgcgtacccc taacttggag gatttgggga cagggaattt gacagatgac ttccaaactg
      841 agtcctactt taagcttacc agtgctctgg agaaatgcaa gatgttgagg agtttgtctg
      901 gattctggga tgcttctcct gtttgcctgt catttatcta ccccctgtgt gctcaactga
      961 caggattgaa cttgagctat gcacccacac ttgatgcttc tgaccttaca aaaatgatta
     1021 gccgctgtgt gaagctccaa cgcctttggg tactggattg tatctcggac aaaggcttgc
     1081 aagtggtggc ctccagttgc aaagacttgc aagaactcag ggtatttcca tcagatttct
     1141 acgtagctgg ttattctgca gtgacagagg agggacttgt tgcagtatcc ttgggctgtc
     1201 caaaactgaa ctcactactg tacttctgtc accaaatgac taatgctgca ctagttactg
     1261 tcgccaagaa ctgtccaaat ttcacacgat tcagactttg tattcttgag ccagggaagc
     1321 ctgatgttgt gacaagccaa ccattagatg aaggctttgg agctattgtt cgtgagtgca
     1381 agggattaca acgtttgtca atatctggtc ttctcacaga caaagttttc atgtatattg
     1441 ggaaatatgc aaaacaactt gagatgcttt ctatagcatt tgctggtgac agtgataagg
     1501 gtatgatgca tgttatgaat ggatgcaaga atttaaggaa actggagata agagatagcc
     1561 cgtttggtga tgctgcactc ttggggaatt ttgctaggta cgagacaatg cgatcccttt
     1621 ggatgtcatc ttgcaatgtc acgttaaagg ggtgccaagt ccttgcgtca aagatgccga
     1681 tgctcaatgt tgaggtcata aatgagcggg atggtagcaa tgaaatggag gaaaaccatg
     1741 gagatctgcc taaagtggag aaattatatg tgtaccgcac aactgctggg gcgagggatg
     1801 atgcaccaaa ttttgttaaa atcctagtcg actccggttc tgctgctagt ggtgaacaaa
     1861 agttgatttc tgaagaagat ttgaacggtg aacaaaagct aatctccgag gaagacttga
     1921 acggtgaaca aaaattaatc tcagaagaag acttgaacgg atccactagc taaggccgcg
     1981 gatctggtta ccactaaacc agcctcaaga acacccgaat ggagtctcta agctacataa
     2041 taccaactta cactttacaa aatgttgtcc cccaaaatgt agccattcgt atctgctcct
     2101 aataaaaaga aagtttcttc acattctaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2161 mccccccccc cccccccctg cagggccaag tcggccggta cccaattcgc cctatagtga
     2221 gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     2281 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     2341 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcaaattgt
     2401 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     2461 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     2521 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     2581 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     2641 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     2701 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     2761 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     2821 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     2881 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2941 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     3001 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3061 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3121 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     3181 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     3241 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3301 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3361 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3421 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3481 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3541 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3601 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3661 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     3721 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     3781 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     3841 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     3901 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     3961 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4021 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4081 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4141 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4201 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4261 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4321 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4381 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4441 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4501 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4561 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4621 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     4681 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     4741 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     4801 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     4861 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     4921 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     4981 tcacacagga aacagctatg accatgatta cgccaagctc g
//
