LOCUS       sequence-110743-         925 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bicistronic expression of Centrin-GFP fusion and Puro resistance 
            gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 925)
  TITLE     Piel Lab Plasmids
REFERENCE   2  (bases 1 to 925)
  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: pIRES Centrin1 GFP Pur
FEATURES             Location/Qualifiers
     source          1..925
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     51..70
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        51..69
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      131..140
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(137..139,140..142,143..853)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(137..139,140..142,143..853)
                     /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(182..203)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(443..462)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     790..811
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 cagagctctc tggctactag agaacccact gcttactggc ttatcgaaat taatacgact
       61 cactataggg agacccaagc ttggtaccga gctcggatcg atatctgcgg cctagctagc
      121 gctaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
      181 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
      241 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
      301 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
      361 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
      421 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
      481 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
      541 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
      601 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
      661 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
      721 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
      781 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
      841 cgagctgtac aagtccggac tcagatctcg agctcaagct tcgaattctg cagtcgacgg
      901 taccatggct tccggcttca agaag
//
