LOCUS       sequence-84972-e         994 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Localization: Protein Expression Vector , Excitation: 505, Emission:
            515. This plasmid encodes Clover H231L based on reference from Bajar
            et. al. Sci Rep. 2016..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 994)
  AUTHORS   Lam AJ, St-Pierre F, Gong Y, Marshall JD, Cranfill PJ, Baird MA, 
            McKeown MR, Wiedenmann J, Davidson MW, Schnitzer MJ, Tsien RY, Lin 
            MZ
  TITLE     Improving FRET dynamic range with bright green and red fluorescent 
            proteins.
  JOURNAL   Nat Methods. 2012 Sep 9. doi: 10.1038/nmeth.2171.
  PUBMED    22961245
REFERENCE   2  (bases 1 to 994)
  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"; journalName: "Nat 
            Methods. 2012 Sep 9. doi: 10.1038/nmeth.2171."
COMMENT     Sequence Label: Clover-pBAD
FEATURES             Location/Qualifiers
     source          1..994
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             94..116
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             136..153
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             157..189
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             202..222
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             229..948
                     /codon_start=1
                     /product="monomeric derivative of the bright green-yellow 
                     fluorescent protein Clover"
                     /label=mClover2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(274..295)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     882..903
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 catttttatc cataagatta gcggatacta ctgacgcttt tatcgcaact ctctactgtt
       61 tctccatacc cgtttttttg ggctagaaat aattttgttt aactttaaga aggagatata
      121 catatgcggg gttctcatca tcatcatcat catggtatgg ctagcatgac tggtggacag
      181 caaatgggtc gggatctgta cgagaacctg tacttccagg gctcgagcat ggtgagcaag
      241 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      301 ggccacaagt tcagcgtccg cggcgagggc gagggcgatg ccaccaacgg caagctgacc
      361 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      421 ttcggctacg gcgtggcctg cttcagccgc taccccgacc acatgaagca gcacgacttc
      481 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatctcttt caaggacgac
      541 ggtacctaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
      601 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
      661 aacttcaaca gccacaacgt ctatatcacg gccgacaagc agaagaacgg catcaaggct
      721 aacttcaaga tccgccacaa cgttgaggac ggcagcgtgc agctcgccga ccactaccag
      781 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagccat
      841 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
      901 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaga attcgaagct
      961 tggctgtttt ggcggatgag agaagatttt cagc
//
