LOCUS       sequence-103915-        1129 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 1129)
  TITLE     Directional cloning vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 1129)
  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            "Unpublished"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..1129
                     /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             182..238
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(383..385,386..1099)
                     /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(383..385,386..1096)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(425..446)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(686..705)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1033..1054
                     /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 tgctactaac ttcagcctgc tcaagcaggc tggagacgtg gaggagaacc ctggacctgt
      241 cgacattacc gattcactgg ccgtcgttct gcaacgccgc gactgggaaa accctggcgt
      301 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agttggcgca atagcgaaga
      361 ggcccgcacc gaccgcgaat tcgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
      421 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
      481 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
      541 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
      601 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
      661 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
      721 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
      781 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
      841 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
      901 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
      961 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     1021 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     1081 ggacgagctg tacaagtaac tcgagtaatc tagataactg atcataatc
//
