LOCUS       sequence-103916-        1073 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  A directional cloning vector of PCR-amplified cDNAs for mammalian 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1073)
  TITLE     Directional cloning vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 1073)
  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            "Unpublished"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: pCMVlac-dirTopo-AU1-aGFP
FEATURES             Location/Qualifiers
     source          1..1073
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    83..99
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             join(344..346,347..1060)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(344..346,347..1057)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(386..407)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(647..666)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     994..1015
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagctttaca ctttattctt
       61 ccggctcgta ttttgtgtgg aattgtgagc ggataacaat ttccggagga ggcccttcac
      121 ctgaagagcg gacggacgag cggatatcgc cgcctaaccc cgctcttcaa agggatccaa
      181 tgacacctac aggtacatcg tcgacattac cgattcactg gccgtcgttc tgcaacgccg
      241 cgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
      301 cagttggcgc aatagcgaag aggcccgcac cgaccgcgaa ttcgtgagca agggcgagga
      361 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
      421 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
      481 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
      541 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
      601 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
      661 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
      721 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
      781 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
      841 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
      901 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
      961 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     1021 cgccgggatc actctcggca tggacgagct gtacaagtaa ctcgagtaat cta
//
