LOCUS       sequence-145670-        1085 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  eF1alpha::puroR::mCherry::T2A::eGFP::MCS::wPRE for assaying effects 
            of C-terminal fusions to eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1085)
  AUTHORS   Arribere JA, Cenik ES, Jain N, Hess GT, Lee CH, Bassik MC, Fire AZ
  TITLE     Translation readthrough mitigation.
  JOURNAL   Nature. 2016 Jun 1;534(7609):719-723. doi: 10.1038/nature18308.
  PUBMED    27281202
REFERENCE   2  (bases 1 to 1085)
  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"; doi: "10"; journalName: 
            "Nature"; date: "2016-06-1- 1"; volume: "534"; issue: "7609"; pages:
            "719-723"
COMMENT     Sequence Label: pJA291
FEATURES             Location/Qualifiers
     source          1..1085
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             86..790
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(228..246)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     496..515
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             800..853
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      860..869
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(911..932)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
       61 cggatcaggt tcaggatctg gtagtgtgag caagggcgag gaggataaca tggccatcat
      121 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
      181 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
      241 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
      301 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
      361 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
      421 ccaggactcc tctctccagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
      481 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga
      541 gcggatgtac cccgaggacg gcgctctgaa aggagagatc aagcagaggc tgaagctgaa
      601 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
      661 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta
      721 caccatcgtg gaacagtacg aacgcgccga gggacggcac tccaccggag gaatggacga
      781 gctgtacaag ggatcgggag agggcagagg aagtctgcta acatgcggtg acgtcgagga
      841 gaatcctggc ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
      901 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtctgg
      961 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     1021 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     1081 cagcc
//
