LOCUS       sequence-142096-         940 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 940)
  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 940)
  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..940
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    93..126
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(173..883,884..886,887..889))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(215..236)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     564..583
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     823..844
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      886..895
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tcaatctttc acaaattttg taatccagag gttgattatc gataagcttg atatcgaatt
       61 gtacctagtg gaaccggaac ccttaaacat gtataacttc gtataatgta tgctatacga
      121 agttattagg tccctcgacg aattcttatt acctgcaggc tccgcggccg cacttgtaca
      181 gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat
      241 cgcgcttctc gttggggtct ttgctcaggg cggactgggt gctcaggtag tggttgtcgg
      301 gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca
      361 cgctgccgtc ctcgatgttg tggcggatct tgaagttcgc cttgatgccg ttcttctgct
      421 tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga
      481 tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc
      541 cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc
      601 gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt
      661 cggggtagcg gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg
      721 gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat
      781 cgccctcgcc ctcgccagac acgctgaact tgtggccgtt tacgtcgccg tccagctcga
      841 ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatg gtggcgaccg
      901 gtgggccagg attctcctcg acgtcaccgc atgttagcag
//
