LOCUS       sequence-251350-        2127 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  BirA* expressed as a protein fusion with MS2 coat protein (MCP) 
            monomer 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 2127)
  TITLE     Jansen lab plasmids
REFERENCE   2  (bases 1 to 2127)
  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 - mcp - egfp - BirA*
FEATURES             Location/Qualifiers
     source          1..2127
                     /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(439..441,442..444,445..1155)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(439..441,442..444,445..1155)
                     /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(484..505)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(745..764)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1092..1113
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             join(1165..1512,1513..1515,1516..2124)
                     /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 atatgatggc ttctaacttt actcagttcg ttctcgtcga caatggcgga
      121 actggcgacg tgactgtcgc cccaagcaac ttcgctaacg ggatcgctga atggatcagc
      181 tctaactcgc gttcacaggc ttacaaagta acctgtagcg ttcgtcagag ctctgcgcag
      241 aatcgcaaat acaccatcaa agtcgaggtg cctaaaggcg cctggcgttc gtacttaaat
      301 atggaactaa ccattccaat tttcgccacg aattccgact gcgagcttat tgttaaggca
      361 atgcaaggtc tcctaaaaga tggaaacccg attccctcag caatcgcagc aaactccggc
      421 atctacgcgg attctagaat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      481 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      541 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
      601 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
      661 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      721 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
      781 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
      841 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
      901 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
      961 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     1021 ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag
     1081 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     1141 gacgagctgt acaagctcga ggacaaggac aacaccgtgc ccctgaagct gatcgccctg
     1201 ctggccaacg gcgagttcca ctctggcgag cagctgggag agaccctggg aatgagcaga
     1261 gccgccatca acaagcacat ccagacactg agagactggg gagtggacgt gttcaccgtg
     1321 cctggcaagg gctacagcct gcctgagcct atccagctgc tgaacgccaa gcagatcctg
     1381 ggacagctgg atggcggaag cgtggccgtg ctgcctgtga tcgactccac caatcagtac
     1441 ctgctggaca gaatcggaga gctgaagtcc ggcgacgcct gcatcgccga gtaccagcag
     1501 gctggcagag gaggcagagg acggaagtgg ttcagcccat tcggagccaa cctgtacctg
     1561 tccatgttct ggagactgga gcagggacct gctgctgcca tcggactgag tctggtgatc
     1621 ggaatcgtga tggccgaggt gctgagaaag ctgggagccg acaaggtgag agtgaagtgg
     1681 cctaatgacc tgtacctcca ggaccgcaag ctggctggca tcctggtgga gctgacaggc
     1741 aagacaggcg atgccgctca gatcgtgatc ggagccggaa tcaacatggc catgagaaga
     1801 gtggaggaga gcgtggtgaa ccagggctgg atcaccctgc aggaggctgg catcaacctg
     1861 gaccggaaca ccctggccgc catgctgatc agagagctga gagccgctct ggagctgttc
     1921 gagcaggagg gactggctcc ttacctgagc agatgggaga agctggacaa cttcatcaac
     1981 agacctgtga agctgatcat cggcgacaag gaaatcttcg gcatctccag aggaatcgac
     2041 aagcagggag ctctgctgct ggagcaggac ggaatcatca agccctggat gggcggagaa
     2101 atctccctga gaagcgcaga gaagtaa
//
