LOCUS       sequence-251344-        2493 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  BirA* expressed as a protein fusion with MS2 coat protein (MCP) 
            dimer and EGFP; 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 2493)
  AUTHORS   Mukherjee J, Hermesh O, Eliscovich C, Nalpas N, Franz-Wachtel M, 
            Macek B, Jansen RP
  TITLE     beta-Actin mRNA interactome mapping by proximity biotinylation.
  JOURNAL   Proc Natl Acad Sci U S A. 2019 Jun 25;116(26):12863-12872. doi: 
            10.1073/pnas.1820737116. Epub 2019 Jun 12.
  PUBMED    31189591
REFERENCE   2  (bases 1 to 2493)
  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"; doi: 
            "10.1073/pnas.1820737116"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2019-06-25- 25"; volume: "116"; issue: "26"; pages: 
            "12863-12872"
COMMENT     Sequence Label: phage - ubc - nls - 2xmcp - egfp - BirA*
FEATURES             Location/Qualifiers
     source          1..2493
                     /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             join(805..807,808..810,811..1521)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(805..807,808..810,811..1521)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(850..871)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1111..1130)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1458..1479
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             join(1531..1878,1879..1881,1882..2490)
                     /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 aggagctgtt caccggggtg
      841 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
      901 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
      961 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     1021 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     1081 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1141 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1201 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     1261 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     1321 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     1381 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     1441 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     1501 ggcatggacg agctgtacaa gctcgaggac aaggacaaca ccgtgcccct gaagctgatc
     1561 gccctgctgg ccaacggcga gttccactct ggcgagcagc tgggagagac cctgggaatg
     1621 agcagagccg ccatcaacaa gcacatccag acactgagag actggggagt ggacgtgttc
     1681 accgtgcctg gcaagggcta cagcctgcct gagcctatcc agctgctgaa cgccaagcag
     1741 atcctgggac agctggatgg cggaagcgtg gccgtgctgc ctgtgatcga ctccaccaat
     1801 cagtacctgc tggacagaat cggagagctg aagtccggcg acgcctgcat cgccgagtac
     1861 cagcaggctg gcagaggagg cagaggacgg aagtggttca gcccattcgg agccaacctg
     1921 tacctgtcca tgttctggag actggagcag ggacctgctg ctgccatcgg actgagtctg
     1981 gtgatcggaa tcgtgatggc cgaggtgctg agaaagctgg gagccgacaa ggtgagagtg
     2041 aagtggccta atgacctgta cctccaggac cgcaagctgg ctggcatcct ggtggagctg
     2101 acaggcaaga caggcgatgc cgctcagatc gtgatcggag ccggaatcaa catggccatg
     2161 agaagagtgg aggagagcgt ggtgaaccag ggctggatca ccctgcagga ggctggcatc
     2221 aacctggacc ggaacaccct ggccgccatg ctgatcagag agctgagagc cgctctggag
     2281 ctgttcgagc aggagggact ggctccttac ctgagcagat gggagaagct ggacaacttc
     2341 atcaacagac ctgtgaagct gatcatcggc gacaaggaaa tcttcggcat ctccagagga
     2401 atcgacaagc agggagctct gctgctggag caggacggaa tcatcaagcc ctggatgggc
     2461 ggagaaatct ccctgagaag cgcagagaag taa
//
