LOCUS       sequence-105891-        1076 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Contains GFP tag for direct live-cell fluorescence imaging of the 
            resofurin ligase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1076)
  AUTHORS   Liu DS, Nivon LG, Richter F, Goldman PJ, Deerinck TJ, Yao JZ, 
            Richardson D, Phipps WS, Ye AZ, Ellisman MH, Drennan CL, Baker D, 
            Ting AY
  TITLE     Computational design of a red fluorophore ligase for site-specific 
            protein labeling in living cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2014 Oct 28;111(43):E4551-9. doi: 
            10.1073/pnas.1404736111. Epub 2014 Oct 13.
  PUBMED    25313043
REFERENCE   2  (bases 1 to 1076)
  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.1404736111"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2014-10-28- 28"; volume: "111"; issue: "43"; pages: "E4551-9"
COMMENT     Sequence Label: pcDNA3-GFP-LplA(AAG)
FEATURES             Location/Qualifiers
     source          1..1076
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     61..80
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        61..79
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      99..108
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             120..137
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(150..152,153..155,156..866)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(150..152,153..155,156..866)
                     /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(195..216)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(456..475)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     803..824
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             876..899
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
       61 taatacgact cactataggg agacccaagc ttgcggccgc caccatggtg cggggttctc
      121 atcatcatca tcatcatggt atggctagca tggtgagcaa gggcgaggag ctgttcaccg
      181 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
      241 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
      301 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
      361 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
      421 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
      481 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
      541 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
      601 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
      661 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
      721 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
      781 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
      841 ctctcggcat ggacgagctg tacaagaagg atccagacta caaggatgac gacgataaga
      901 tgagcaccct gagactgctg atcagcgaca gctacgaccc ctggttcaac ctggccgtgg
      961 ccgagtgcat cttcagacag atgcccgcca cccagagagt gctgttcctg tggagaaacg
     1021 ccgacaccgt ggtgatcggc agagcccaga acccctggaa ggagtgcaac accaga
//
