LOCUS       sequence-436663-        5300 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector BS2 in nucleus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5300)
  AUTHORS   Pani S, Qiu T, Kentala K, Azizi SA, Dickinson BC
  TITLE     Bioorthogonal masked acylating agents for proximity-dependent RNA 
            labelling.
  JOURNAL   Nat Chem. 2024 Apr 9. doi: 10.1038/s41557-024-01493-1.
  PUBMED    38594368
REFERENCE   2  (bases 1 to 5300)
  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: "Nat Chem. 
            2024 Apr 9. doi: 10.1038/s41557-024-01493-1."
COMMENT     Sequence Label: V5-BS2-NLS (Nucleoplasmin)
FEATURES             Location/Qualifiers
     source          1..5300
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(400..441)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     regulatory      441..450
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(492..511)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          494..626
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(762..965)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(791..811)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        966..1345
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    1536..1819
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     rep_origin      1935..2523
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2424..2443
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2695..3489)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MLEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFAKLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2806..2825)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3416..3435
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     polyA_signal    3888..4009
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3925..3944)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3979..3998
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4046..4064
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4182..4229)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 caatgtaacg acaatgactt ctccctgtgc cgcaagtttt gatccgtcat acaatggctc
       61 actgcccgct cctagataaa aagcgcctcc gtgaatccac accatgacag gaagattttt
      121 acttggggtg tcaggcgcaa atacattgac atacaagcaa tcctcggact ggcggggcag
      181 ctcagtatac gaaagtgaca gcaaatcaga cggctgcggg caaatagagc cgtacgctgt
      241 ggcatcaagc acatcttccc acacttcagg cggctcaggt gctttaaaac gccattgtcc
      301 gacaggcggc ttggcatagg ggatgccttt ccacttatgt acgccgtttt ccgttgtgcc
      361 ttttactttg ccgtattgag tcgttactat ttgatgagtg gtgctgtcca ggcccagcag
      421 ggggttgggg atgggcttgc ccatggtggc atactctagc cttaagagct gtaattgaac
      481 tgggagtgga cacctgtgga gagaaaggca aagtggatgt cagtaagacc aataggtgcc
      541 tatcagaaac gcaagagtct tctctgtctc gacaagccca gtttctattg gtctccttaa
      601 acctgtcttg taaccttgat acttacctgc ccagtgcctc acgaccaact tctgcagctt
      661 aagttcgaga ctgttgtgtc agaagcactg actgcgttag caatttaact gtgataaact
      721 accgcaataa agcttctagt gatctgacgg ttcactaaac cagctctgct tatatagacc
      781 tcccaccgta cacgcctacc gcccatttgc gtcaacgggg cggggttatt acgacatttt
      841 ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga
      901 cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tggtgtactg ccaaaaccgc
      961 atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccgt
     1021 aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg
     1081 ggcggacttg gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac
     1141 tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt
     1201 attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt
     1261 tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat
     1321 taataactag tcaataatca atgccaacat ggcggtcata ttggacatga gccaatataa
     1381 atgtacatat tatgatatag atacaacgta tgcaatggcc aatagccaat attgatttat
     1441 gctatataac caatgaataa tatggctaat ggccaatatt gaagatctcg atcctctacg
     1501 ccggacgcat cgtggccgga gtactgcggt ttatctcgat ggctacgagg gcagacagta
     1561 agtggattta ccataatccc ttaattgtac gcaccgctaa aacgcgttca gcgcgatcac
     1621 ggcagcagac aggtaaaaat ggcaacaaac cacccgaaaa actgccgcaa tcgcgcctga
     1681 taaattttaa ccgtatgaat acctatgcaa ccagagggta caggccacat tacccccact
     1741 taatccactg aagctgccat ttttcatggt ttcaccatcc cagcgaaggg ccatccagcg
     1801 tgcgttcctg tatttccgga ctataaaagg atctaggtga agatcctttt tgataatctc
     1861 atgaccaaaa tcacttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     1921 atcaaaggat cttcttgaaa tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     1981 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2041 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     2101 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2161 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2221 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2281 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2341 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2401 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2461 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2521 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     2581 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     2641 gctgataccg gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa
     2701 gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta
     2761 aagcacgagg aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc
     2821 caacgctatg tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga
     2881 aaagcggcca ttttccacca tgatattcgg caagcaggca tcgccatgag tcacgacgag
     2941 atcctcacca tccggcatac gcgccttgag cttggcgaac agttcggctg gcgcgagccc
     3001 ctgatgctct tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtgcgtgc
     3061 tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg
     3121 cagccgccgc attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga
     3181 caggagatcc tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac
     3241 aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc
     3301 ctcgtcctgc agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg
     3361 cccctgcgct gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca
     3421 gtcatagccg aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg
     3481 ttcaagcatg ctaaacgacc gtcatcctgt ctcttgatca gatcttgatc ccctgcgcca
     3541 tcagatcctt ggcggcaaga aagccatcca gtttactttg cagggcttcc caaccttacc
     3601 agagggcgcc ccagctggca attcttttga agctcacgct gccgcaagca ctcagggcgc
     3661 aagggctgct aaaggaagcg gaacacgtag aaagccagtc cgcagaaacg gtgctgaccc
     3721 cggatgaatg tcagctactg ggctatctgg acaagggaaa acgcaagcgc aaagaggatc
     3781 cattagaatt cgattttacc acatttgtag aggttttact tgcttaaaaa aacctcccac
     3841 atctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     3901 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3961 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgct
     4021 cgaagcggcc gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc
     4081 agcttccttt cgggctttgt tagcagccgg atcagcttgc atgcctgcag gtcgactcta
     4141 gaggatcccc gggtaccgag cccgaattcg tttaaacatt actttttctt ttttgcctgg
     4201 ccggcctttt tcgtggccgc cggccttttg gatccttctc cttttgaagg gaatagcttc
     4261 tgcctttttt cagattcggg atcgttttcg atcgtaatct ctgagtctaa aatcagcgtc
     4321 tctctcgttt cttcatgata cgcaggccaa ttcacagctt cggtgcttgg gtttcctgtt
     4381 ttggcgaacg tgatccacgc tgattgtatc gtgtgagaaa gctgtttcac ctcatccgta
     4441 atctccgctt ttgccattcg ttccaatccg tccagatttc caaagacaaa aggaagctct
     4501 aatgcgtgaa acgctttatt gtacggcggc ttcttcgggt gccaatcgaa cctgtacatc
     4561 cagacagggg cgtaatgaga ctgtgcggat gcataggcga cggcagggcg ccaaaataat
     4621 aaatcagtca tcatgtgaat ttggctttcc agagaacgcg gatacaaatc ggcaactttc
     4681 tctgccagcg gcttccctag taaatactcg agcgctgcat caagcgtttc ctgagaatga
     4741 acgtctgaat ccggggtgaa aaataaatat ccttcatcac gggttgttcc aattaatagc
     4801 ggaataccgg aagcagcccc ttctgcgatc gctttttctg gttcttcagg cagcgttttc
     4861 ggatcaaggg cgggctggaa gaacagctga aagatatttt ctttttctgc aatccgaagc
     4921 tgatcagccg cttttagcaa atcttccgca gaaaccgtat gcaatttatc cagttggccc
     4981 tcgttaatcc caaggacctg taaaaaggct gccgaggtgc tcgccgcttg ttctttcgtc
     5041 atcgttcgag aagcgccgct ttccatgatt gctttctgga acaggccttt tgccgcaggc
     5101 atagcaagca gcgcggcaat gctcatcccg ccggcggatt ctccaaatac tgttacgtta
     5161 tcgggatcac cgccaaacgc tgaaatattc tctcgcaccc atttcagcgc ggcggcttgg
     5221 tctaaaagcc caaggttatc agaatacgcc tcattaaatg aagacaagtg caaaaagcca
     5281 aacggcccca gccgatagtt
//
