LOCUS       sequence-165337-        2161 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 2161)
  AUTHORS   Provost E, Rhee J, Leach SD
  TITLE     Viral 2A peptides allow expression of multiple proteins from a 
            single ORF in transgenic zebrafish embryos.
  JOURNAL   Genesis. 2007 Oct . 45(10):625-9.
  PUBMED    17941043
REFERENCE   2  (bases 1 to 2161)
  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: "Genesis. 
            2007 Oct . 45(10):625-9."
COMMENT     Sequence Label: pEF6V5:eGFP-CAAX-2A-mCherry
FEATURES             Location/Qualifiers
     source          1..2161
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          37..240
                     /label=EF-1-alpha intron A
                     /note="truncated intron upstream of the start codon of 
                     human EF-1-alpha"
     primer_bind     197..217
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        266..284
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     356..373
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     357..373
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        380..398
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(477..479,480..482,483..1193)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPTLVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(522..543)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(783..802)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1130..1151
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1242..1298
                     /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             1299..2009
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1444..1462)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1712..1731
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             2101..2142
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
ORIGIN
        1 gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag tacgtcgtct
       61 ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg ggtggagact
      121 gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct ttttgagttt
      181 ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct tccatttcag
      241 gtgtcgtgag gaattagctt ggtactaata cgactcacta tagggagacc caagctggct
      301 aggtaagctt ggtaccgagc tcggatccac tagtccagtg tggtggaatt gcccttgtaa
      361 aacgacggcc agtgaattgt aatacgactc actatagggc gaattgggcc ctctagatgc
      421 atgctcgagc ggccgccagt gtgatggata tctgcagaat tcgcccttcg ctctccatgg
      481 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccac cctggtcgag ctggacggcg
      541 acgtaaacgg ccacaagttc agcgtgtctg gcgagggcga gggcgatgcc acctacggca
      601 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
      661 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
      721 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
      781 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
      841 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
      901 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
      961 tcaaggcgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     1021 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1081 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1141 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagaagaaga
     1201 aaaagaagtc aaagacaaag tgtgtaatta tgggatccgg agccacgaac ttctctctgt
     1261 taaagcaagc aggagacgtg gaagaaaacc ccggtcctat ggtgagcaag ggcgaggagg
     1321 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga
     1381 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     1441 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc
     1501 ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact
     1561 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     1621 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga
     1681 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct
     1741 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc
     1801 agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg
     1861 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct
     1921 cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca
     1981 ccggcggcat ggacgagctg tacaagtaaa tcgattctcg ctcttcgctc tgccaagggc
     2041 aattctgcag atatccagca cagtggcggc cgctcgagtc tagagggccc gcggttcgaa
     2101 ggtaagccta tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg tcatcatcac
     2161 c
//
