LOCUS       sequence-103914-        1101 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 1101)
  TITLE     Directional cloning vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 1101)
  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            "Unpublished"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-P2A-aGFP
FEATURES             Location/Qualifiers
     source          1..1101
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     polyA_signal    131..252
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(142..161)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     196..215
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(379..1092,1093..1095))
                     /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             complement(join(382..1092,1093..1095))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(424..445)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     773..792
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1032..1053
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
       61 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
      121 atttacgcgt taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
      181 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
      241 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
      301 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcag
      361 ttatctagat tactcgagtt acttgtacag ctcgtccatg ccgagagtga tcccggcggc
      421 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc
      481 ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
      541 gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt
      601 gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt tgtggctgtt
      661 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt
      721 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta
      781 gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga
      841 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta
      901 ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa
      961 cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt
     1021 gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc
     1081 ctcgcccttg ctcacgaatt c
//
