LOCUS       sequence-431776-        5378 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector BS2 in nucleolus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5378)
  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 5378)
  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-NIK3x (Nucleolus)
FEATURES             Location/Qualifiers
     source          1..5378
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    102..385
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     enhancer        576..955
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        956..1159
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1110..1130
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          1295..1427
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1410..1429
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1471..1480
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1480..1521
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3235..3253)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3290..3411
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3301..3320)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3355..3374
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             3810..4604
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3864..3883)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4474..4493
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4776..5364)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4856..4875)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
       61 gagattatca aaaaggatct tcacctagat ccttttatag tccggaaata caggaacgca
      121 cgctggatgg cccttcgctg ggatggtgaa accatgaaaa atggcagctt cagtggatta
      181 agtgggggta atgtggcctg taccctctgg ttgcataggt attcatacgg ttaaaattta
      241 tcaggcgcga ttgcggcagt ttttcgggtg gtttgttgcc atttttacct gtctgctgcc
      301 gtgatcgcgc tgaacgcgtt ttagcggtgc gtacaattaa gggattatgg taaatccact
      361 tactgtctgc cctcgtagcc atcgagataa accgcagtac tccggccacg atgcgtccgg
      421 cgtagaggat cgagatcttc aatattggcc attagccata ttattcattg gttatatagc
      481 ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat
      541 ttatattggc tcatgtccaa tatgaccgcc atgttggcat tgattattga ctagttatta
      601 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      661 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      721 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      781 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc
      841 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      901 acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      961 gcggttttgg cagtacacca atgggcgtgg atagcggttt gactcacggg gatttccaag
     1021 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     1081 aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga
     1141 ggtctatata agcagagctg gtttagtgaa ccgtcagatc actagaagct ttattgcggt
     1201 agtttatcac agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt
     1261 aagctgcaga agttggtcgt gaggcactgg gcaggtaagt atcaaggtta caagacaggt
     1321 ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc gtttctgata
     1381 ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg tccactccca
     1441 gttcaattac agctcttaag gctagagtat gccaccatgg gcaagcccat ccccaacccc
     1501 ctgctgggcc tggacagcac cactcatcaa atagtaacga ctcaatacgg caaagtaaaa
     1561 ggcacaacgg aaaacggcgt acataagtgg aaaggcatcc cctatgccaa gccgcctgtc
     1621 ggacaatggc gttttaaagc acctgagccg cctgaagtgt gggaagatgt gcttgatgcc
     1681 acagcgtacg gctctatttg cccgcagccg tctgatttgc tgtcactttc gtatactgag
     1741 ctgccccgcc agtccgagga ttgcttgtat gtcaatgtat ttgcgcctga caccccaagt
     1801 aaaaatcttc ctgtcatggt gtggattcac ggaggcgctt tttatctagg agcgggcagt
     1861 gagccattgt atgacggatc aaaacttgcg gcacagggag aagtcattgt cgttacattg
     1921 aactatcggc tggggccgtt tggctttttg cacttgtctt catttaatga ggcgtattct
     1981 gataaccttg ggcttttaga ccaagccgcc gcgctgaaat gggtgcgaga gaatatttca
     2041 gcgtttggcg gtgatcccga taacgtaaca gtatttggag aatccgccgg cgggatgagc
     2101 attgccgcgc tgcttgctat gcctgcggca aaaggcctgt tccagaaagc aatcatggaa
     2161 agcggcgctt ctcgaacgat gacgaaagaa caagcggcga gcacctcggc agccttttta
     2221 caggtccttg ggattaacga gggccaactg gataaattgc atacggtttc tgcggaagat
     2281 ttgctaaaag cggctgatca gcttcggatt gcagaaaaag aaaatatctt tcagctgttc
     2341 ttccagcccg cccttgatcc gaaaacgctg cctgaagaac cagaaaaagc gatcgcagaa
     2401 ggggctgctt ccggtattcc gctattaatt ggaacaaccc gtgatgaagg atatttattt
     2461 ttcaccccgg attcagacgt tcattctcag gaaacgcttg atgcagcgct cgagtattta
     2521 ctagggaagc cgctggcaga gaaagttgcc gatttgtatc cgcgttctct ggaaagccaa
     2581 attcacatga tgactgattt attattttgg cgccctgccg tcgcctatgc atccgcacag
     2641 tctcattacg cccctgtctg gatgtacagg ttcgattggc acccgaagaa gccgccgtac
     2701 aataaagcgt ttcacgcatt agagcttcct tttgtctttg gaaatctgga cggattggaa
     2761 cgaatggcaa aagcggagat tacggatgag gtgaaacagc tttctcacac gatacaatca
     2821 gcgtggatca cgttcgccaa aacaggaaac ccaagcaccg aagctgtgaa ttggcctgcg
     2881 tatcatgaag aaacgagaga gacgctgatt ttagactcag agattacgat cgaaaacgat
     2941 cccgaatctg aaaaaaggca gaagctattc ccttcaaaag gagaaggatc ctcgaattca
     3001 agaatgagaa agaagcgaaa aaagaagctt cgaatcctgc gcatgagaaa gaagcgcaag
     3061 aagaagctcc gcatcctcag gatgcgcaag aaacgcaaga agaagctgcg catcctgtga
     3121 tgtttaaacg aattcgggct cggtacccgg ggatcctcta gagtcgacct gcaggcatgc
     3181 aagctgatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
     3241 agcaataact agcataaccc cttggggcgg ccgcttcgag cagacatgat aagatacatt
     3301 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     3361 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     3421 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt tttaagcaag
     3481 taaaacctct acaaatgtgg taaaatcgaa ttctaatgga tcctctttgc gcttgcgttt
     3541 tcccttgtcc agatagccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga
     3601 ctggctttct acgtgttccg cttcctttag cagcccttgc gccctgagtg cttgcggcag
     3661 cgtgagcttc aaaagaattg ccagctgggg cgccctctgg taaggttggg aagccctgca
     3721 aagtaaactg gatggctttc ttgccgccaa ggatctgatg gcgcagggga tcaagatctg
     3781 atcaagagac aggatgacgg tcgtttcgca tgcttgaaca agatggattg cacgcaggtt
     3841 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3901 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3961 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4021 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4081 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4141 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4201 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgcactcgg atggaagccg
     4261 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4321 tcgccaggct caaggcgcgt atgccggatg gtgaggatct cgtcgtgact catggcgatg
     4381 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     4441 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4501 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4561 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4621 cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg
     4681 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4741 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4801 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4861 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4921 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4981 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5041 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5101 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5161 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     5221 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5281 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5341 acgcgcagaa aaaaaggatt tcaagaagat cctttgat
//
