LOCUS       sequence-257125-        5160 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  in vitro transcription for mRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5160)
  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 5160)
  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..5160
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(60..78)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(61..78)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(96..112)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(96..112)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(109..131)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    120..136
                     /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(144..150,151..168,169..174))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    189..210
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(327..344)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(498..1086)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(578..597)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1257..2048,2049..2117))
                     /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     1880..1899
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2118..2222)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2290..2308
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2346..2368)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2468..2487
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2681..2703
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2695..2712
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2696..2712
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2719..2738
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2719..2737
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           2769..2811
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     regulatory      2845..2854
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2851..3558
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2996..3014)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3264..3283
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             3595..4983
                     /codon_start=1
                     /product="mouse RING E3 ubiquitin ligase that can be used 
                     together with antibodies to degrade intracellular proteins 
                     (Clift et al., 2018)"
                     /label=TRIM21
                     /note="binds to the Fc domain of antibodies"
                     /translation="MSLEKMWEEVTCSICLDPMVEPMSIECGHCFCKECIFEVGKNGGS
                     SCPECRQQFLLRNLRPNRHIANMVENLKQIAQNTKKSTQETHCMKHGEKLHLFCEEDGQ
                     ALCWVCAQSGKHRDHTRVPIEEAAKVYQEKIHVALEKLRKGKELAEKMEMDLTMQRTDW
                     KRNIDTQKSRIHAEFALQNSLLAQEEQRQLQRLEKDQREYLRLLGKKEAELAEKNQALQ
                     ELISELERRIRGSELELLQEVRIILERSGSWNLDTLDIDAPDLTSTCPVPGRKKMLRTC
                     WVHITLDRNTANSWLIISKDRRQVRMGDTHQNVSDNKERFSNYPMVLGAQRFSSGKMYW
                     EVDVTQKEAWDLGVCRDSVQRKGQFSLSPENGFWTIWLWQDSYEAGTSPQTTLHIQVPP
                     CQIGIFVDYEAGVVSFYNITDHGSLIYTFSECVFAGPLRPFFNVGFNYSGGNAAPLKLC
                     PLKM"
     3'UTR           5027..5152
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
ORIGIN
        1 cccccccccc ctgcaggcgc gccgccggcg atttaaatta attaatctag cttgagtatt
       61 ctatagtgtc acctaaatag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
      121 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
      181 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
      241 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
      301 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
      361 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
      421 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
      481 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
      541 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
      601 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
      661 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
      721 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
      781 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
      841 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
      901 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
      961 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     1021 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     1081 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     1141 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     1201 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     1261 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     1321 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     1381 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     1441 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     1501 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     1561 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     1621 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     1681 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     1741 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     1801 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     1861 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     1921 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     1981 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     2041 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     2101 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     2161 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     2221 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     2281 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     2341 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     2401 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     2461 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     2521 cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac
     2581 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     2641 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     2701 acgacggcca gtgaattgta atacgactca ctatagggcg aattaattcg agctcggtac
     2761 ccagcttgct tgttcttttt gcagaagctc agaataaacg ctcaactttg gcagatcaat
     2821 tccccggtgc tagcgctacc ggtcgccacc atggtgagca agggcgagga ggataacatg
     2881 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
     2941 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     3001 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
     3061 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
     3121 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
     3181 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
     3241 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
     3301 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg
     3361 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
     3421 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac
     3481 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
     3541 atggacgagc tgtacaagtc cggactcaga tctcgagctc aagcttcgaa ttcaatgtct
     3601 ctggaaaaga tgtgggagga ggtcacctgt tctatctgcc tggatcccat ggtggagcct
     3661 atgagtatcg aatgtggcca ttgcttttgc aaggaatgca tttttgaagt tgggaagaat
     3721 gggggcagtt catgtcccga gtgccggcaa cagtttctgc tccgaaacct caggcccaat
     3781 agacatatag ccaacatggt ggaaaacctt aaacagatag cccagaatac caagaagagt
     3841 acccaggaaa cgcactgcat gaagcatgga gagaagcttc acctattctg tgaggaagat
     3901 gggcaggccc tttgctgggt gtgtgcccag tctgggaaac accgggacca caccagggtc
     3961 cctattgaag aggctgctaa ggtataccag gagaagatcc acgtggcttt agaaaaactg
     4021 agaaagggga aagagttggc cgagaagatg gaaatggatc tcacgatgca aagaacagac
     4081 tggaagagga acattgacac ccagaagtcg aggattcacg cagagttcgc acttcagaat
     4141 agcttgctgg ctcaggagga gcagaggcag ctgcagaggc tggagaagga tcaaagggag
     4201 tacctgagac tcctggggaa gaaggaggct gagctggctg agaagaacca ggccctgcag
     4261 gagctgatct cagagctgga gaggaggatt cgtggttcag agctggagct actgcaggag
     4321 gtgaggatca tcctggaaag gagtggatcc tggaacctgg acacgttaga tattgacgcc
     4381 ccagacctaa caagcacatg ccctgtgcca gggcggaaga agatgctgag gacgtgttgg
     4441 gttcatatta ctctggatcg caacaccgcc aactcatggc tcatcatctc aaaggatcgg
     4501 agacaagtga ggatgggaga cacccatcag aacgtgtctg acaataagga gaggtttagt
     4561 aattacccca tggtgctagg tgcccagaga ttctcctctg ggaagatgta ctgggaggta
     4621 gatgtgactc aaaaggaggc ctgggatctg ggggtttgca gagattctgt tcagaggaaa
     4681 gggcagtttt cactcagtcc cgagaatggc ttctggacca tttggttatg gcaagacagc
     4741 tatgaggctg gtaccagtcc tcagaccacc ctccacattc aagtacctcc atgccaaatt
     4801 gggatctttg tggactatga ggctggcgtt gtctccttct acaacataac tgaccatggc
     4861 tccctcattt acaccttctc ggagtgtgtt tttgctggac ctctgcgacc tttcttcaat
     4921 gttggtttca attatagtgg gggaaatgca gcgcctctaa agctctgtcc actaaagatg
     4981 tgagcggccg ctctagataa atgatagctt gatctggtta ccactaaacc agcctcaaga
     5041 acacccgaat ggagtctcta agctacataa taccaactta cactttacaa aatgttgtcc
     5101 cccaaaatgt agccattcgt atctgctcct aataaaaaga aagtttcttc acattctaaa
//
