LOCUS       sequence-436847-        5321 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector BS2 in mitocondria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5321)
  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 5321)
  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: Mito-V5-BS2 (Mitochondrial matrix)
FEATURES             Location/Qualifiers
     source          1..5321
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    165..286
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(202..221)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     256..275
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     323..341
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(1929..1970)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2090..2109)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          2092..2224
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(2360..2563)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2389..2409)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2564..2943
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    3134..3417
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     rep_origin      3533..4121
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4022..4041
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4293..5087)
                     /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(4404..4423)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5014..5033
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 atgaatgtca gctactgggc tatctggaca agggaaaacg caagcgcaaa gaggatccat
       61 tagaattcga ttttaccaca tttgtagagg ttttacttgc ttaaaaaaac ctcccacatc
      121 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
      181 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
      241 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgctcga
      301 agcggccgcc ccaaggggtt atgctagtta ttgctcagcg gtggcagcag ccaactcagc
      361 ttcctttcgg gctttgttag cagccggatc agcttgcatg cctgcaggtc gactctagag
      421 gatccccggg taccgagccc gaattcgttt aaacattagg atccttctcc ttttgaaggg
      481 aatagcttct gccttttttc agattcggga tcgttttcga tcgtaatctc tgagtctaaa
      541 atcagcgtct ctctcgtttc ttcatgatac gcaggccaat tcacagcttc ggtgcttggg
      601 tttcctgttt tggcgaacgt gatccacgct gattgtatcg tgtgagaaag ctgtttcacc
      661 tcatccgtaa tctccgcttt tgccattcgt tccaatccgt ccagatttcc aaagacaaaa
      721 ggaagctcta atgcgtgaaa cgctttattg tacggcggct tcttcgggtg ccaatcgaac
      781 ctgtacatcc agacaggggc gtaatgagac tgtgcggatg cataggcgac ggcagggcgc
      841 caaaataata aatcagtcat catgtgaatt tggctttcca gagaacgcgg atacaaatcg
      901 gcaactttct ctgccagcgg cttccctagt aaatactcga gcgctgcatc aagcgtttcc
      961 tgagaatgaa cgtctgaatc cggggtgaaa aataaatatc cttcatcacg ggttgttcca
     1021 attaatagcg gaataccgga agcagcccct tctgcgatcg ctttttctgg ttcttcaggc
     1081 agcgttttcg gatcaagggc gggctggaag aacagctgaa agatattttc tttttctgca
     1141 atccgaagct gatcagccgc ttttagcaaa tcttccgcag aaaccgtatg caatttatcc
     1201 agttggccct cgttaatccc aaggacctgt aaaaaggctg ccgaggtgct cgccgcttgt
     1261 tctttcgtca tcgttcgaga agcgccgctt tccatgattg ctttctggaa caggcctttt
     1321 gccgcaggca tagcaagcag cgcggcaatg ctcatcccgc cggcggattc tccaaatact
     1381 gttacgttat cgggatcacc gccaaacgct gaaatattct ctcgcaccca tttcagcgcg
     1441 gcggcttggt ctaaaagccc aaggttatca gaatacgcct cattaaatga agacaagtgc
     1501 aaaaagccaa acggccccag ccgatagttc aatgtaacga caatgacttc tccctgtgcc
     1561 gcaagttttg atccgtcata caatggctca ctgcccgctc ctagataaaa agcgcctccg
     1621 tgaatccaca ccatgacagg aagattttta cttggggtgt caggcgcaaa tacattgaca
     1681 tacaagcaat cctcggactg gcggggcagc tcagtatacg aaagtgacag caaatcagac
     1741 ggctgcgggc aaatagagcc gtacgctgtg gcatcaagca catcttccca cacttcaggc
     1801 ggctcaggtg ctttaaaacg ccattgtccg acaggcggct tggcataggg gatgcctttc
     1861 cacttatgta cgccgttttc cgttgtgcct tttactttgc cgtattgagt cgttactatt
     1921 tgatgagtgg tgctgtccag gcccagcagg gggttgggga tgggcttgcc gtgagcacgt
     1981 acgcatacag atgtagagat agcacgcttc cctaccagag aaaatacacg tgtagccagc
     2041 atggtggcat actctagcct taagagctgt aattgaactg ggagtggaca cctgtggaga
     2101 gaaaggcaaa gtggatgtca gtaagaccaa taggtgccta tcagaaacgc aagagtcttc
     2161 tctgtctcga caagcccagt ttctattggt ctccttaaac ctgtcttgta accttgatac
     2221 ttacctgccc agtgcctcac gaccaacttc tgcagcttaa gttcgagact gttgtgtcag
     2281 aagcactgac tgcgttagca atttaactgt gataaactac cgcaataaag cttctagtga
     2341 tctgacggtt cactaaacca gctctgctta tatagacctc ccaccgtaca cgcctaccgc
     2401 ccatttgcgt caacggggcg gggttattac gacattttgg aaagtcccgt tgattttggt
     2461 gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa
     2521 accgctatcc acgcccattg gtgtactgcc aaaaccgcat caccatggta atagcgatga
     2581 ctaatacgta gatgtactgc caagtaggaa agtcccgtaa ggtcatgtac tgggcataat
     2641 gccaggcggg ccatttaccg tcattgacgt caataggggg cggacttggc atatgataca
     2701 cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga
     2761 aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt
     2821 cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat
     2881 atgggctatg aactaatgac cccgtaattg attactatta ataactagtc aataatcaat
     2941 gccaacatgg cggtcatatt ggacatgagc caatataaat gtacatatta tgatatagat
     3001 acaacgtatg caatggccaa tagccaatat tgatttatgc tatataacca atgaataata
     3061 tggctaatgg ccaatattga agatctcgat cctctacgcc ggacgcatcg tggccggagt
     3121 actgcggttt atctcgatgg ctacgagggc agacagtaag tggatttacc ataatccctt
     3181 aattgtacgc accgctaaaa cgcgttcagc gcgatcacgg cagcagacag gtaaaaatgg
     3241 caacaaacca cccgaaaaac tgccgcaatc gcgcctgata aattttaacc gtatgaatac
     3301 ctatgcaacc agagggtaca ggccacatta cccccactta atccactgaa gctgccattt
     3361 ttcatggttt caccatccca gcgaagggcc atccagcgtg cgttcctgta tttccggact
     3421 ataaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc acttaacgtg
     3481 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgaaatc
     3541 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     3601 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     3661 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3721 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3781 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3841 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3901 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3961 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4021 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4081 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4141 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4201 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccggt tgggcgtcgc
     4261 ttggtcggtc atttcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag
     4321 aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc
     4381 cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg
     4441 tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg
     4501 atattcggca agcaggcatc gccatgagtc acgacgagat cctcaccatc cggcatacgc
     4561 gccttgagct tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca
     4621 tcctgatcga caagaccggc ttccatccga gtgcgtgctc gctcgatgcg atgtttcgct
     4681 tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc
     4741 atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact
     4801 tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa
     4861 ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg
     4921 gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac
     4981 acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc
     5041 acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caagcatgct aaacgaccgt
     5101 catcctgtct cttgatcaga tcttgatccc ctgcgccatc agatccttgg cggcaagaaa
     5161 gccatccagt ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat
     5221 tcttttgaag ctcacgctgc cgcaagcact cagggcgcaa gggctgctaa aggaagcgga
     5281 acacgtagaa agccagtccg cagaaacggt gctgaccccg g
//
