LOCUS       sequence-72892-e        1248 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian tdTomato activation reporter for NM with 
            GTCCCCTCCACCCCACAGTG protospacer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1248)
  AUTHORS   Esvelt KM, Mali P, Braff JL, Moosburner M, Yaung SJ, Church GM
  TITLE     Orthogonal Cas9 proteins for RNA-guided gene regulation and editing.
  JOURNAL   Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681.
  PUBMED    24076762
REFERENCE   2  (bases 1 to 1248)
  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: "Nat 
            Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681."
COMMENT     Sequence Label: M-tdTom-NM
FEATURES             Location/Qualifiers
     source          1..1248
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        53..91
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     88..112
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      126..135
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             135..158
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             186..884
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL
                     VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ
                     ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH
                     LFLYGMDELYK"
     primer_bind     complement(316..334)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     584..603
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     778..801
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    1084..1165
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1121..1140)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1176..1193)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tagggataac agggtaatag tgtcccctcc accccacagt gggaggattc gaggtaggcg
       61 tgtacggtgg gaggcctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag
      121 aattcgccac catggactac aaggatgacg acgataaaac ttccggtggc ggactgggtt
      181 ccaccgtgag caagggcgag gaggtcatca aagagttcat gcgcttcaag gtgcgcatgg
      241 agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
      301 agggcaccca gaccgccaag ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg
      361 acatcctgtc cccccagttc atgtacggct ccaaggcgta cgtgaagcac cccgccgaca
      421 tccccgatta caagaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
      481 tcgaggacgg cggtctggtg accgtgaccc aggactcctc cctgcaggac ggcacgctga
      541 tctacaaggt gaagatgcgc ggcaccaact tcccccccga cggccccgta atgcagaaga
      601 agaccatggg ctgggaggcc tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg
      661 gcgagatcca ccaggccctg aagctgaagg acggcggcca ctacctggtg gagttcaaga
      721 ccatctacat ggccaagaag cccgtgcaac tgcccggcta ctactacgtg gacaccaagc
      781 tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgag cgctccgagg
      841 gccgccacca cctgttcctg tacggcatgg acgagctgta caagtaagaa ttcgagctcg
      901 gtacccgggg atcctctagt cagctgacgc gtgctagcgc ggccgcatcg ataagcttgt
      961 cgacgatatc tctagaggat cataatcagc cataccacat ttgtagaggt tttacttgct
     1021 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     1081 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     1141 acaaataaag catttttttc actgcctcga gcttcctcgc tcactgactc gctgcgctcg
     1201 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaata
//
