LOCUS       sequence-82629-e         877 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cre dependent expression of ChIEF kinetic variant E162A/T198C with 
            physically uncoupled dTomato fluorophore .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 877)
  TITLE     ChIEF and oChIEF AAV vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 877)
  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            "Unpublished"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: 
            AAV-EF1a-DIO-ChIEF(E162A/T198C)-P2A-dTomato-WPRE-BGHpA
FEATURES             Location/Qualifiers
     source          1..877
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    complement(33..66)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(117..150)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(168..869)
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(251..274)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(449..468)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     718..736
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
ORIGIN
        1 gaggtaccgg atcctctaga gtcgactccg gaataacttc gtataggata ctttatacga
       61 agttatgcag aatggtagct ggattgtagc tgctattagc aatatgaaac ctcttaataa
      121 cttcgtatag catacattat acgaagttat ggcgcgccgt cgacttactt gtacagctcg
      181 tccatgccgt acaggaacag gtggtggcgg ccctcggagc gctcgtactg ttccacgatg
      241 gtgtagtcct cgttgtggga ggtgatgtcc agcttggtgt ccacgtagta gtagccgggc
      301 agttgcacgg gcttcttggc catgtagatg gtcttgaact ccaccaggta gtggccgccg
      361 tccttcagct tcagggcctg gtggatctcg cccttcagca cgccgtcgcg ggggtacagg
      421 cgctcggtgg aggcctccca gcccatggtc ttcttctgca ttacggggcc gtcggggggg
      481 aagttggtgc cgcgcatctt caccttgtag atcagcgtgc cgtcctgcag ggaggagtcc
      541 tgggtcacgg tcaccagacc gccgtcctcg aagttcatca cgcgctccca cttgaagccc
      601 tcggggaagg acagcttctt gtaatcgggg atgtcggcgg ggtgcttcac gtacgccttg
      661 gagccgtaca tgaactgggg ggacaggatg tcccaggcga agggcagggg gccgcccttg
      721 gtcaccttca gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg
      781 atctcgaact cgtggccgtt catggagccc tccatgcgca ccttgaagcg catgaactct
      841 ttgatgacct cctcgccctt gctcaccatt ggcccgg
//
