LOCUS       sequence-128869-         869 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Retroviral expression of Centrin 2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 869)
  TITLE     Cheeseman Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 869)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCB99
FEATURES             Location/Qualifiers
     source          1..869
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      21..30
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(27..29,30..32,33..743)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(27..29,30..32,33..743)
                     /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(72..93)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(333..352)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     680..701
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             789..809
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             822..866
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
ORIGIN
        1 gtggtggtac gctaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
       61 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
      121 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
      181 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
      241 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
      301 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
      361 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
      421 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
      481 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
      541 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
      601 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
      661 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
      721 tcggcatgga cgagctgtac aagtccggag gggatcggtg gagctccacc ggtggcggcc
      781 gctctagaga gaatctttat tttcagggcg ccgccaaatt caaagaaacc gctgctgcta
      841 aattcgaacg ccagcacatg gacagcgga
//
