LOCUS       sequence-110746-         933 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bicistronic expression of GFP-Centrin fusion and Hygro resistance 
            gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 933)
  TITLE     Piel Lab Plasmids
REFERENCE   2  (bases 1 to 933)
  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 GFP Centrin1  Hygro
FEATURES             Location/Qualifiers
     source          1..933
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     42..61
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        42..60
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      129..138
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(135..137,138..140,141..851)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(135..137,138..140,141..851)
                     /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(180..201)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(441..460)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     788..809
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 tggctactag agaacccact gccttactgg cttatcgaaa ttaatacgac tcactatagg
       61 gagacccaag cttggtaccg agctcggatc tgtacaggcc ttaagccggc gcgctagcgc
      121 taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
      181 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
      241 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
      301 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
      361 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
      421 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
      481 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
      541 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
      601 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
      661 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
      721 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
      781 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
      841 agctgtacaa gtccggactc agatctcgag ctcaagcttc gaattctgca gtcgacggta
      901 ccatggcttc cggcttcaag aagccagcgc tgc
//
