LOCUS       sequence-72887-e        1247 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian tdTomato activation reporter for SP with 
            GTCCCCTCCACCCCACAGTG protospacer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1247)
  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 1247)
  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-SP
FEATURES             Location/Qualifiers
     source          1..1247
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        52..90
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     87..111
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      125..134
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             134..157
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             185..883
                     /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(315..333)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     583..602
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     777..800
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    1083..1164
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1120..1139)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1175..1192)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tagggataac agggtaatag tgtcccctcc accccacagt gcgggaaacg aggtaggcgt
       61 gtacggtggg aggcctatat aagcagagct cgtttagtga accgtcagat cgcctggaga
      121 attcgccacc atggactaca aggatgacga cgataaaact tccggtggcg gactgggttc
      181 caccgtgagc aagggcgagg aggtcatcaa agagttcatg cgcttcaagg tgcgcatgga
      241 gggctccatg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
      301 gggcacccag accgccaagc tgaaggtgac caagggcggc cccctgccct tcgcctggga
      361 catcctgtcc ccccagttca tgtacggctc caaggcgtac gtgaagcacc ccgccgacat
      421 ccccgattac aagaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
      481 cgaggacggc ggtctggtga ccgtgaccca ggactcctcc ctgcaggacg gcacgctgat
      541 ctacaaggtg aagatgcgcg gcaccaactt cccccccgac ggccccgtaa tgcagaagaa
      601 gaccatgggc tgggaggcct ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg
      661 cgagatccac caggccctga agctgaagga cggcggccac tacctggtgg agttcaagac
      721 catctacatg gccaagaagc ccgtgcaact gcccggctac tactacgtgg acaccaagct
      781 ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgagc gctccgaggg
      841 ccgccaccac ctgttcctgt acggcatgga cgagctgtac aagtaagaat tcgagctcgg
      901 tacccgggga tcctctagtc agctgacgcg tgctagcgcg gccgcatcga taagcttgtc
      961 gacgatatct ctagaggatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     1021 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     1081 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     1141 caaataaagc atttttttca ctgcctcgag cttcctcgct cactgactcg ctgcgctcgg
     1201 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaata
//
