LOCUS       sequence-251356-        2484 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  BirA* expressed as a protein fusion with MS2 coat protein (MCP) 
            dimer and mCherry; to visualize any MS2 tagged mRNAs and to identify
            the RNA proximal proteome by their biotinylation. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2484)
  TITLE     Jansen lab plasmids
REFERENCE   2  (bases 1 to 2484)
  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"
COMMENT     Sequence Label: phage - ubc - nls - 2xmcp - mCherry - BirA*
FEATURES             Location/Qualifiers
     source          1..2484
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             7..27
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             31..57
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             31..57
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             58..69
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             805..1512
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(950..968)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1218..1237
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             join(1522..1869,1870..1872,1873..2481)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
ORIGIN
        1 atgggcccaa aaaagaaaag aaaagttggc tacccctacg acgtgcccga ctacgccatc
       61 gaaggccgcc atatgctagc cgttaaaatg gcttctaact ttactcagtt cgttctcgtc
      121 gacaatggcg gaactggcga cgtgactgtc gccccaagca acttcgctaa cgggatcgct
      181 gaatggatca gctctaactc gcgttcacag gcttacaaag taacctgtag cgttcgtcag
      241 agctctgcgc agaatcgcaa atacaccatc aaagtcgagg tgcctaaagg cgcctggcgt
      301 tcgtacttaa atatggaact aaccattcca attttcgcca cgaattccga ctgcgagctt
      361 attgttaagg caatgcaagg tctcctaaaa gatggaaacc cgattccctc agcaatcgca
      421 gcaaactccg gcatctacgc catggcttct aactttactc agttcgttct cgtcgacaat
      481 ggcggaactg gcgacgtgac tgtcgcccca agcaacttcg ctaacgggat cgctgaatgg
      541 atcagctcta actcgcgttc acaggcttac aaagtaacct gtagcgttcg tcagagctct
      601 gcgcagaatc gcaaatacac catcaaagtc gaggtgccta aaggcgcctg gcgttcgtac
      661 ttaaatatgg aactaaccat tccaattttc gccacgaatt ccgactgcga gcttattgtt
      721 aaggcaatgc aaggtctcct aaaagatgga aacccgattc cctcagcaat cgcagcaaac
      781 tccggcatct acgcggattc tagaatggtg agcaagggcg aggaggataa catggccatc
      841 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
      901 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
      961 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
     1021 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
     1081 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     1141 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     1201 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc
     1261 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
     1321 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
     1381 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
     1441 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     1501 gagctgtaca agctcgagga caaggacaac accgtgcccc tgaagctgat cgccctgctg
     1561 gccaacggcg agttccactc tggcgagcag ctgggagaga ccctgggaat gagcagagcc
     1621 gccatcaaca agcacatcca gacactgaga gactggggag tggacgtgtt caccgtgcct
     1681 ggcaagggct acagcctgcc tgagcctatc cagctgctga acgccaagca gatcctggga
     1741 cagctggatg gcggaagcgt ggccgtgctg cctgtgatcg actccaccaa tcagtacctg
     1801 ctggacagaa tcggagagct gaagtccggc gacgcctgca tcgccgagta ccagcaggct
     1861 ggcagaggag gcagaggacg gaagtggttc agcccattcg gagccaacct gtacctgtcc
     1921 atgttctgga gactggagca gggacctgct gctgccatcg gactgagtct ggtgatcgga
     1981 atcgtgatgg ccgaggtgct gagaaagctg ggagccgaca aggtgagagt gaagtggcct
     2041 aatgacctgt acctccagga ccgcaagctg gctggcatcc tggtggagct gacaggcaag
     2101 acaggcgatg ccgctcagat cgtgatcgga gccggaatca acatggccat gagaagagtg
     2161 gaggagagcg tggtgaacca gggctggatc accctgcagg aggctggcat caacctggac
     2221 cggaacaccc tggccgccat gctgatcaga gagctgagag ccgctctgga gctgttcgag
     2281 caggagggac tggctcctta cctgagcaga tgggagaagc tggacaactt catcaacaga
     2341 cctgtgaagc tgatcatcgg cgacaaggaa atcttcggca tctccagagg aatcgacaag
     2401 cagggagctc tgctgctgga gcaggacgga atcatcaagc cctggatggg cggagaaatc
     2461 tccctgagaa gcgcagagaa gtaa
//
