LOCUS       sequence-92932-e         790 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses His-tagged EGFP-mOrange fusion protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 790)
  AUTHORS   Tome JM, Ozer A, Pagano JM, Gheba D, Schroth GP, Lis JT
  TITLE     Comprehensive analysis of RNA-protein interactions by 
            high-throughput sequencing-RNA affinity profiling.
  JOURNAL   Nat Methods. 2014 Jun;11(6):683-8. doi: 10.1038/nmeth.2970. Epub 
            2014 May 8.
  PUBMED    24809628
REFERENCE   2  (bases 1 to 790)
  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: 
            "10.1038/nmeth.2970"; journalName: "Nat Methods"; date: "2014-06"; 
            volume: "11"; issue: "6"; pages: "683-8"
COMMENT     Sequence Label: EGFP-mOrange pHis-//
FEATURES             Location/Qualifiers
     source          1..790
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     43..62
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        43..61
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    62..86
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             101..123
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             143..160
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             182..202
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(215..217,218..220,221..733)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(260..281)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(521..540)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga ctcactatag
       61 gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg
      121 agatatacat atgtcgtact accatcacca tcaccatcac gattacgata tcccaacgac
      181 cgaaaacctg tattttcagg gcgccatgga tccgatggtg agcaagggcg aggagctgtt
      241 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
      301 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
      361 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
      421 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
      481 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
      541 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
      601 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
      661 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
      721 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
      781 cggcgacggc
//
