LOCUS       sequence-436846-        5288 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector BS2 in cytoplasm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5288)
  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 5288)
  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-NES (Cytosol)
FEATURES             Location/Qualifiers
     source          1..5288
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        74..377
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        378..581
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     532..552
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          717..849
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     832..851
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      893..902
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             902..943
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2420..2446
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(2567..2585)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2622..2743
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2633..2652)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2687..2706
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             3142..3936
                     /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(3196..3215)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3806..3825
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4108..4696)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4188..4207)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    4812..5095
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
       61 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      121 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      181 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      241 tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      301 attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag
      361 tcatcgctat taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt
      421 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      481 accaaaatca acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg
      541 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
      601 tcactagaag ctttattgcg gtagtttatc acagttaaat tgctaacgca gtcagtgctt
      661 ctgacacaac agtctcgaac ttaagctgca gaagttggtc gtgaggcact gggcaggtaa
      721 gtatcaaggt tacaagacag gtttaaggag accaatagaa actgggcttg tcgagacaga
      781 gaagactctt gcgtttctga taggcaccta ttggtcttac tgacatccac tttgcctttc
      841 tctccacagg tgtccactcc cagttcaatt acagctctta aggctagagt atgccaccat
      901 gggcaagccc atccccaacc ccctgctggg cctggacagc accactcatc aaatagtaac
      961 gactcaatac ggcaaagtaa aaggcacaac ggaaaacggc gtacataagt ggaaaggcat
     1021 cccctatgcc aagccgcctg tcggacaatg gcgttttaaa gcacctgagc cgcctgaagt
     1081 gtgggaagat gtgcttgatg ccacagcgta cggctctatt tgcccgcagc cgtctgattt
     1141 gctgtcactt tcgtatactg agctgccccg ccagtccgag gattgcttgt atgtcaatgt
     1201 atttgcgcct gacaccccaa gtaaaaatct tcctgtcatg gtgtggattc acggaggcgc
     1261 tttttatcta ggagcgggca gtgagccatt gtatgacgga tcaaaacttg cggcacaggg
     1321 agaagtcatt gtcgttacat tgaactatcg gctggggccg tttggctttt tgcacttgtc
     1381 ttcatttaat gaggcgtatt ctgataacct tgggctttta gaccaagccg ccgcgctgaa
     1441 atgggtgcga gagaatattt cagcgtttgg cggtgatccc gataacgtaa cagtatttgg
     1501 agaatccgcc ggcgggatga gcattgccgc gctgcttgct atgcctgcgg caaaaggcct
     1561 gttccagaaa gcaatcatgg aaagcggcgc ttctcgaacg atgacgaaag aacaagcggc
     1621 gagcacctcg gcagcctttt tacaggtcct tgggattaac gagggccaac tggataaatt
     1681 gcatacggtt tctgcggaag atttgctaaa agcggctgat cagcttcgga ttgcagaaaa
     1741 agaaaatatc tttcagctgt tcttccagcc cgcccttgat ccgaaaacgc tgcctgaaga
     1801 accagaaaaa gcgatcgcag aaggggctgc ttccggtatt ccgctattaa ttggaacaac
     1861 ccgtgatgaa ggatatttat ttttcacccc ggattcagac gttcattctc aggaaacgct
     1921 tgatgcagcg ctcgagtatt tactagggaa gccgctggca gagaaagttg ccgatttgta
     1981 tccgcgttct ctggaaagcc aaattcacat gatgactgat ttattatttt ggcgccctgc
     2041 cgtcgcctat gcatccgcac agtctcatta cgcccctgtc tggatgtaca ggttcgattg
     2101 gcacccgaag aagccgccgt acaataaagc gtttcacgca ttagagcttc cttttgtctt
     2161 tggaaatctg gacggattgg aacgaatggc aaaagcggag attacggatg aggtgaaaca
     2221 gctttctcac acgatacaat cagcgtggat cacgttcgcc aaaacaggaa acccaagcac
     2281 cgaagctgtg aattggcctg cgtatcatga agaaacgaga gagacgctga ttttagactc
     2341 agagattacg atcgaaaacg atcccgaatc tgaaaaaagg cagaagctat tcccttcaaa
     2401 aggagaagga tccctgcagc tgcctcccct ggagcgcctg accctggact aatgtttaaa
     2461 cgaattcggg ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagctgat
     2521 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     2581 ctagcataac cccttggggc ggccgcttcg agcagacatg ataagataca ttgatgagtt
     2641 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     2701 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     2761 tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct
     2821 ctacaaatgt ggtaaaatcg aattctaatg gatcctcttt gcgcttgcgt tttcccttgt
     2881 ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt
     2941 ctacgtgttc cgcttccttt agcagccctt gcgccctgag tgcttgcggc agcgtgagct
     3001 tcaaaagaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac
     3061 tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagatc tgatcaagag
     3121 acaggatgac ggtcgtttag catgcttgaa caagatggat tgcacgcagg ttctccggcc
     3181 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3241 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3301 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     3361 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3421 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3481 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3541 gaccaccaag cgaaacatcg catcgagcga gcacgcactc ggatggaagc cggtcttgtc
     3601 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccaag
     3661 ctcaaggcgc gtatgccgga tggtgaggat ctcgtcgtga ctcatggcga tgcctgcttg
     3721 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     3781 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     3841 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     3901 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     3961 ccgaccaagc gacgcccaac cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4021 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4081 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4141 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4201 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4261 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4321 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4381 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4441 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4501 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     4561 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4621 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4681 aaaaaaagga tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4741 cgaaaactca cgttaagtga ttttggtcat gagattatca aaaaggatct tcacctagat
     4801 ccttttatag tccggaaata caggaacgca cgctggatgg cccttcgctg ggatggtgaa
     4861 accatgaaaa atggcagctt cagtggatta agtgggggta atgtggcctg taccctctgg
     4921 ttgcataggt attcatacgg ttaaaattta tcaggcgcga ttgcggcagt ttttcgggtg
     4981 gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc tgaacgcgtt ttagcggtgc
     5041 gtacaattaa gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa
     5101 accgcagtac tccggccacg atgcgtccgg cgtagaggat cgagatcttc aatattggcc
     5161 attagccata ttattcattg gttatatagc ataaatcaat attggctatt ggccattgca
     5221 tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc
     5281 atgttggc
//
