LOCUS       sequence-32563-e         859 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 859)
  TITLE     New vectors Aug. 2010
REFERENCE   2  (bases 1 to 859)
  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: pLenti CMV/TO GFP-MDC1 (779-2)
FEATURES             Location/Qualifiers
     source          1..859
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    12..30
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    71..95
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      136..145
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(142..144,145..147,148..858)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(142..144,145..147,148..858)
                     /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(187..208)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(448..467)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     795..816
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 tgatagagat ctccctatca gtgatagaga tcgtcgacta gtccagtgtg gtggaattct
       61 gcagatatca acaagtttgt acaaaaaagc aggctccacc atgggaacca attcagtcga
      121 ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
      181 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      241 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
      301 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
      361 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
      421 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
      481 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
      541 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
      601 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
      661 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
      721 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
      781 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
      841 atggacgagc tgtacaagt
//
