LOCUS       sequence-234219-         989 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Template for C-terminal PCR tagging of mammalian genes (without 
            selection) with TAP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 989)
  AUTHORS   Fueller J, Herbst K, Meurer M, Gubicza K, Kurtulmus B, Knopf JD, 
            Kirrmaier D, Buchmuller BC, Pereira G, Lemberg MK, Knop M
  TITLE     CRISPR-Cas12a-assisted PCR tagging of mammalian genes.
  JOURNAL   J Cell Biol. 2020 Jun 1;219(6). pii: 151766. doi: 
            10.1083/jcb.201910210.
  PUBMED    32406907
REFERENCE   2  (bases 1 to 989)
  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: "J Cell 
            Biol."; date: "2020-06-1"; volume: "219(6). pii: 151766. doi"; 
            pages: " 10.1083/jcb.201910210"
COMMENT     Sequence Label: pMaCTag-24
FEATURES             Location/Qualifiers
     source          1..989
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             41..118
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             146..166
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             206..379
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     CDS             380..553
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNGAQAPK"
     polyA_signal    593..714
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(630..649)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     684..703
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(721..754)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     761..781
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     932..951
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 tcgtacgtca ggtggaggag gtagtggcgg aggcggatcc aagagaagat ggaaaaagaa
       61 tttcatagcc gtctcagcag ccaaccgctt taagaaaatc tcatcctccg gggcacttga
      121 ttatgatatt ccaactactg ctagcgagaa tttgtatttt cagggtgagc tcaaaaccgc
      181 ggctcttgcg caacacgatg aagccgtgga caacaaattc aacaaagaac aacaaaacgc
      241 gttctatgag atcttacatt tacctaactt aaacgaagaa caacgaaacg ccttcatcca
      301 aagtttaaaa gatgacccaa gccaaagcgc taacctttta gcagaagcta aaaagctaaa
      361 tgatgctcag gcgccgaaag tagacaacaa attcaacaaa gaacaacaaa acgcgttcta
      421 tgagatctta catttaccta acttaaacga agaacaacga aacgccttca tccaaagttt
      481 aaaagatgac ccaagccaaa gcgctaacct tttagcagaa gctaaaaagc taaatggtgc
      541 tcaggcgccg aaagtagacg cgaattccgc ggggaagtca acctgaacta gtaacttgtt
      601 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
      661 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttagtcgat
      721 ataacttcgt atagcataca ttatacgaag ttatgtcgag gagggcctat ttcccatgat
      781 tccttcatat ttgcatatac gatacaaggc tgttagagag ataattggaa ttaatttgac
      841 tgtaaacaca aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta
      901 gtttgcagtt ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa
      961 gtatttcgat ttcttggctt tatatatct
//
