LOCUS       sequence-226729-        7915 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 7915)
  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 7915)
  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_Kdm5b_MUT
FEATURES             Location/Qualifiers
     source          1..7915
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     432..451
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     685..702
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    819..840
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(855..860,861..878,879..885)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    893..909
                     /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     898..920
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     917..933
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     917..933
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        954..972
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     5'UTR           997..1039
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             1069..1134
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1510..1521
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     3'UTR           5841..5966
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(6056..6075)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6057..6075)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6082..6099)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6082..6098)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6091..6113)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(6240..6695)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6377..6398)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6589..6608
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6722..6826
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6827..6895,6896..7687)
                     /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(7045..7064)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
       61 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
      121 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
      181 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
      241 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
      301 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
      361 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      421 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      481 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      541 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      601 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
      661 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
      721 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      781 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
      841 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      901 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct cgaaattaac
      961 cctcactaaa gggaacaaaa gctggagcta agcttgcttg ttctttttgc agaagctcag
     1021 aataaacgct caactttggc agatctgaat tcctgcagcc cggggatgga ctacaaagac
     1081 catgacggtg attataaaga tcatgatatc gattacaagg atgacgatga caagggatct
     1141 gcaagaagag catctgtgga attcctgcag cccgggtcca tggagccggc caccacgctg
     1201 cccccaggcc cgcgcccggc gctgcccctc gggggcccgg gcccgctggg agagttcctg
     1261 cctccccccg agtgtccggt cttcgagccg agctgggaag agttcgcgga ccccttcgct
     1321 ttcatccaca agatccggcc catagccgag cagactggga tctgtaaggt gcggccgccg
     1381 ccggattggc agccaccatt cgcttgtgat gtcgataaac ttcatttcac cccgcggatc
     1441 cagaggctga atgagctgga ggcccaaaca cgtgtaaaat tgaatttcct ggaccagatt
     1501 gcaaagtatt gggagctaca aggaagtact ctgaagattc cccacgtgga gagaaagatc
     1561 ttagacttgt tccagctaaa taagttagtc gcagaagaag gtgggtttgc agttgtttgc
     1621 aaggatagaa aatggaccaa aattgccacc aagatgggtt ttgctcctgg taaagctgtg
     1681 ggctcacata tcaggggaca ttatgagcga atactcaatc catacaattt gttcctgtct
     1741 ggagacagtc tgaggtgtct tcagaagcct aacctgacct ccgacacaaa ggacaaggag
     1801 tacaaacccc atgatattcc ccagaggcag tctgtgcagc ccgcagaaac ctgtcctccg
     1861 gcccgccgag caaagcgcat gagagcagag gctatgaata ttaaaattga gccagaagag
     1921 gcaacagaag ccaggactca taatctgaga cgccgaatgg gttgcacaac tccaaaatgg
     1981 gaaaatgaga aagaaatgaa aagcaccatc aagcaagagc ccactgagaa gaaagactgc
     2041 gagttagaaa gtgagaagga gaagcccaag agccgggcca agaaaacggc cactgcggtg
     2101 gacctgtatg tgtgcctctt atgtggcagt ggcaatgacg aggaccggct cctactgtgt
     2161 gacggctgtg atgacagtta ccacaccttc tgcctggttc cccctctcca tgacgttccc
     2221 aagggagact ggaggtgtcc taagtgtttg gctcaagaat gtaataagcc acaagaagct
     2281 tttggctttg aacaagcagc aagggactat acacttcgta cgtttggaga aatggcagat
     2341 gcattcaaat ctgattactt caacatgcca gtccatatgg ttcccacaga gctggtggag
     2401 aaggagtttt ggcggctcgt aagcactatt gaagaagacg tcacggtgga gtatggagct
     2461 gatattgctt caaaagagtt tggcagtggc tttcctgttc gagatgggaa aatcaagatt
     2521 tctcctgagg aagaggaata cctggatagt ggctggaatt tgaacaacat gccagtgatg
     2581 gagcagtctg tccttgctca tattactgct gatatatgtg gcatgaaact cccctggttg
     2641 tatgtgggca tgtgcttttc ttcattctgt tgggccattg aagaccactg gagctattca
     2701 atcaactacc tacactgggg tgagccaaaa acctggtatg gagttccagg ctacgctgct
     2761 gagcagctag aaaacgtgat gaagaagctc gctcctgagc tctttgtgtc ccagccggac
     2821 ctgctccacc agctggtgac catcatgaac cccaacactc tgatgactca tgaagtgcct
     2881 gtttatcgaa cgaatcagtg tgctggggag tttgtaatta cgtttccaag agcctaccac
     2941 agtggcttca accaaggttt taattttgcc gaggctgtta acttctgcac tgttgattgg
     3001 ctgccattgg gccgccagtg tgtggagcat tatcgcttgc tgcaccgtta ttgtgtcttt
     3061 tcccacgatg aaatgatatg caaaatggct tctaaggctg atgtattaga cgttgtggta
     3121 gcgtcaactg ttcagaagga catggccatc atgatcgagg acgagaaagc tttaagggaa
     3181 actgttcgga aattgggagt aattgattct gaaagaatgg attttgagct gttgccagat
     3241 gatgagcggc agtgtataaa atgcaaaact acatgcttca tgtctgccat ctcctgctct
     3301 tgtaaacctg gcctacttgt ttgcctacat catgtgaaag aattgtgttc ttgccccccg
     3361 tataagtata acctgcggta tagatacact ctggatgatc tctatccaat gatgaatgca
     3421 ttaaagcttc gagcagagtc ttacaatgaa tgggccttaa atgtgaatga agccttggaa
     3481 gcaaagatca ataagaagaa aagtcttgtc agctttaaag ctttgattga agaatcagaa
     3541 atgaagaaat tcccagacaa tgatcttttg cgtcaccttc gactagtcac gcaggatgca
     3601 gaaaagtgtg cctctgttgc ccagcagttg cttaatggca aaaggcaaac aaggtatcga
     3661 tctggtggag gaaagtctca gaatcagctg acggtgaatg aacttagaca gtttgtgact
     3721 cagctgtatg ctctgccgtg cgtccttagt cagacaccgt tgctaaagga tctcttgaat
     3781 cgtgtggaag atttccaaca gcaaagccaa aaactactgt ctgaggaaat gcctagcgct
     3841 gccgagctac aggagttgct ggatgtcagc tttgaatttg atgttgagct tccacagctc
     3901 actgagatgc gcattcgctt agagcaggcc cgctggctgg aagaggttca gcaggcttgc
     3961 ctggactcca gctccctttc tttggatgat atgagacgac tcatagactt aggggtgggg
     4021 ctggctccat attctgctgt ggagaaagct atggcccgcc tgcaggagct gctaacagtg
     4081 tcagaacact gggacgacaa agccaagagt ctcctcagag caagaccacg gcattctttg
     4141 agtagccttg ctacagcagt caaagagatg gaggagatcc ccgcctacct gcccaatggt
     4201 acagtcctga aggattctgt gcaaagagcc agagactggg ttcaggatgt ggatgcattg
     4261 caggcaggag gacgtgtgcc agtgttagag acactcattg aacttgttgc aagaggccga
     4321 tctatcccag tacatctcaa ttctttacca agattggaaa tgttagtagc tgaagttcat
     4381 gcatggaaag aatgtgctgc taaaacattc ttgcctgaga attctacata ttctctgtta
     4441 gaggttcttt gccctcggtg tgacattggc cttttggggc tgaaaaggaa acagagaaag
     4501 ttgaaggagc ccttgccgag tggaaagaag agaagcacca agttagagag cctgagcgat
     4561 ctggagaggg ctctgatgga gagcaaagag actgcggcag ctatggcaac tcttggggaa
     4621 gctcgcctac gagagatgga agctttgcag tctctcagat ttgccaatga agagaaattg
     4681 ctgtcacctg tccaagactt agagatgaaa gtctgcttgt gtcagaagac tccagccact
     4741 cccatgattc agtgtgaact ctgcagggat gctttccata ctagttgtgt ggcagcaccc
     4801 agtatttcac aaagctcccg aatctggctt tgtcctcact gtcggaggtc agagaagcct
     4861 ccattagaga aaattctgcc tctgctggcc tccctacagc gtatacgagt tcgtcttcct
     4921 gaaggagatg cactccgata catgattgaa agaaccgtga actggcaaca tagagcccag
     4981 cagctgcttt cttcagggaa tctaaaactt gtgcaagacc aagtgggttc tggactgttg
     5041 tctagcagat ggccagcctc agcaggacag gcatcagcaa cagacaaggt gtctcagcct
     5101 cctggcacaa catccttttc attgcccgat gactgggaca acagaacctc atatttacac
     5161 tctcccttct ctactggaca gagttgtctc cccctccatg gtttgagtcc agaagtgaat
     5221 gaattgctga tggaagccca gctgctccag gtatcccttc ctgaaattca ggagctttat
     5281 cagactctac ttactaagcc aagctctgtt cagcaagctg accgaagttc accagttaga
     5341 tccagcagtg agaagaacga ttgccttcga gggaaacgag atgcaatcaa cagtcctgag
     5401 aggaaactga aaaggcgccc agaaagagaa ggccttccta gcgagcggtg ggatcgagtt
     5461 aaacacatgc ggacccccca aaagaagaaa atcaaactga gtcaccccaa ggacatggac
     5521 agtttcaagt tagaaagaga gcgtagctat gacttagtgc gtaatgctga aactcattcc
     5581 ctgccctcag acacatccta ctccgaacag gaggactctg aggatgaaga tgccatctgt
     5641 ccagctgtga gctgcctgca gccagaagga gatgaggtgg actgggtcca gtgtgatggc
     5701 agctgcaatc agtggtttca ccaggtctgt gttggtgtct ctccagagat ggcagagaag
     5761 gaagactaca tctgtgtgcg ctgtactggg aaggacgcac caagccgaaa gtaaagcggc
     5821 cgcggatctg gttaccacta aaccagcctc aagaacaccc gaatggagtc tctaagctac
     5881 ataataccaa cttacacttt acaaaatgtt gtcccccaaa atgtagccat tcgtatctgc
     5941 tcctaataaa aagaaagttt cttcacattc taaaaaaaaa aaaaaaaaaa aaaacccccc
     6001 cccccccccc cccccccccc cccctgcagg gccaagtcgg ccggtaccca attcgcccta
     6061 tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     6121 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     6181 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggca
     6241 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     6301 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     6361 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     6421 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     6481 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     6541 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     6601 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     6661 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     6721 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     6781 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     6841 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     6901 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     6961 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     7021 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     7081 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     7141 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     7201 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     7261 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     7321 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     7381 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     7441 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     7501 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     7561 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     7621 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     7681 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     7741 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     7801 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     7861 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgcta
//
