LOCUS       sequence-142100-        1115 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  eF1alpha::puroR::mCherry::T2A::eGFP::d4ODC::wPRE has d4ODC degron.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1115)
  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 1115)
  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: pJA318
FEATURES             Location/Qualifiers
     source          1..1115
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    94..127
                     /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(162..284,285..290,291..293,294..1004,
                     1005..1007,1008..1010))
                     /codon_start=1
                     /product="EGFP destabilized by a mutated version of 
                     residues 422-461 of mouse ornithine decarboxylase, giving 
                     an in vivo half-life of ~4 hours"
                     /label=d4EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKCGRSHGFPPEVEEQDDGALPMSCAQESGMDRHPAACASARIN
                     V"
     primer_bind     complement(336..357)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     685..704
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     944..965
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1007..1016
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1023..1076)
                     /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"
ORIGIN
        1 gtcaatcttt cacaaatttt gtaatccaga ggttgattat cgataagctt gatatcgaat
       61 tgtacctagt ggaaccggaa cccttaaaca tgtataactt cgtataatgt atgctatacg
      121 aagttattag gtccctcgac gaattcttat tacctgcagg cctacacatt gatcctagca
      181 gaagcacagg ctgcagggtg acggtccatc ccgctctcct gggcacaaga catgggcagc
      241 gcgccatcat cctgctcctc cacctccggc gggaagccat ggctgcggcc gcacttgtac
      301 agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga
      361 tcgcgcttct cgttggggtc tttgctcagg gcggactggg tgctcaggta gtggttgtcg
      421 ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc
      481 acgctgccgt cctcgatgtt gtggcggatc ttgaagttcg ccttgatgcc gttcttctgc
      541 ttgtcggcca tgatatagac gttgtggctg ttgtagttgt actccagctt gtgccccagg
      601 atgttgccgt cctccttgaa gtcgatgccc ttcagctcga tgcggttcac cagggtgtcg
      661 ccctcgaact tcacctcggc gcgggtcttg tagttgccgt cgtccttgaa gaagatggtg
      721 cgctcctgga cgtagccttc gggcatggcg gacttgaaga agtcgtgctg cttcatgtgg
      781 tcggggtagc ggctgaagca ctgcacgccg taggtcaggg tggtcacgag ggtgggccag
      841 ggcacgggca gcttgccggt ggtgcagatg aacttcaggg tcagcttgcc gtaggtggca
      901 tcgccctcgc cctcgccaga cacgctgaac ttgtggccgt ttacgtcgcc gtccagctcg
      961 accaggatgg gcaccacccc ggtgaacagc tcctcgccct tgctcaccat ggtggcgacc
     1021 ggtgggccag gattctcctc gacgtcaccg catgttagca gacttcctct gccctctccc
     1081 gatcccttgt acagctcgtc cattcctccg gtgga
//
