LOCUS       sequence-67405-e        1243 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  TF reporter for gRNA-AAVS1_T2 (RNA-guided dTomato expression).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1243)
  AUTHORS   Mali P, Aach J, Stranges PB, Esvelt KM, Moosburner M, Kosuri S, Yang
            L, Church GM
  TITLE     CAS9 transcriptional activators for target specificity screening and
            paired nickases for cooperative genome engineering.
  JOURNAL   Nat Biotechnol. 2013 Aug 1. doi: 10.1038/nbt.2675.
  PUBMED    23907171
REFERENCE   2  (bases 1 to 1243)
  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 
            Biotechnol. 2013 Aug 1. doi: 10.1038/nbt.2675."
COMMENT     Sequence Label: reporter-gT2
FEATURES             Location/Qualifiers
     source          1..1243
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        48..86
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     83..107
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      121..130
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             130..153
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             181..879
                     /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(311..329)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     579..598
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     773..796
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    1079..1160
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1116..1135)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1171..1188)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tagggataac agggtaatag tggggccact agggacagga ttggcgaggt aggcgtgtac
       61 ggtgggaggc ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagaattc
      121 gccaccatgg actacaagga tgacgacgat aaaacttccg gtggcggact gggttccacc
      181 gtgagcaagg gcgaggaggt catcaaagag ttcatgcgct tcaaggtgcg catggagggc
      241 tccatgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
      301 acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc
      361 ctgtcccccc agttcatgta cggctccaag gcgtacgtga agcaccccgc cgacatcccc
      421 gattacaaga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
      481 gacggcggtc tggtgaccgt gacccaggac tcctccctgc aggacggcac gctgatctac
      541 aaggtgaaga tgcgcggcac caacttcccc cccgacggcc ccgtaatgca gaagaagacc
      601 atgggctggg aggcctccac cgagcgcctg tacccccgcg acggcgtgct gaagggcgag
      661 atccaccagg ccctgaagct gaaggacggc ggccactacc tggtggagtt caagaccatc
      721 tacatggcca agaagcccgt gcaactgccc ggctactact acgtggacac caagctggac
      781 atcacctccc acaacgagga ctacaccatc gtggaacagt acgagcgctc cgagggccgc
      841 caccacctgt tcctgtacgg catggacgag ctgtacaagt aagaattcga gctcggtacc
      901 cggggatcct ctagtcagct gacgcgtgct agcgcggccg catcgataag cttgtcgacg
      961 atatctctag aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     1021 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     1081 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     1141 taaagcattt ttttcactgc ctcgagcttc ctcgctcact gactcgctgc gctcggtcgt
     1201 tcggctgcgg cgagcggtat cagctcactc aaaggcggta ata
//
