LOCUS       sequence-178238-        4404 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of dead/inactive increased fidelity SpCas9-HF1-VP64-6xHis
            in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4404)
  AUTHORS   Kulcsar PI, Talas A, Huszar K, Ligeti Z, Toth E, Weinhardt N, Fodor 
            E, Welker E
  TITLE     Crossing enhanced and high fidelity SpCas9 nucleases to optimize 
            specificity and cleavage.
  JOURNAL   Genome Biol. 2017 Oct 6;18(1):190. doi: 10.1186/s13059-017-1318-8.
  PUBMED    28985763
REFERENCE   2  (bases 1 to 4404)
  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: "Genome 
            Biol."; date: "2017-10-6"; volume: "18(1):190. doi"; pages: " 
            10.1186/s13059-017-1318-8"
COMMENT     Sequence Label: pET-dSpCas9-HF1-VP64-6xHis
FEATURES             Location/Qualifiers
     source          1..4404
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             4111..4131
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4138..4203
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4234..4383
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             4384..4401
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 atggataaga aatactcaat aggcttagct atcggcacaa atagcgtcgg atgggcggtg
       61 atcactgatg aatataaggt tccgtctaaa aagttcaagg ttctgggaaa tacagaccgc
      121 cacagtatca aaaaaaatct tataggggct cttttatttg acagtggaga gacagcggaa
      181 gcgactcgtc tcaaacggac agctcgtaga aggtatacac gtcggaagaa tcgtatttgt
      241 tatctacagg agattttttc aaatgagatg gcgaaagtag atgatagttt ctttcatcga
      301 cttgaagagt cttttttggt ggaagaagac aagaagcatg aacgtcatcc tatttttgga
      361 aatatagtag atgaagttgc ttatcatgag aaatatccaa ctatctatca tctgcgaaaa
      421 aaattggtag attctactga taaagcggat ttgcgcttaa tctatttggc cttagcgcat
      481 atgattaagt ttcgtggtca ttttttgatt gagggagatt taaatcctga taatagtgat
      541 gtggacaaac tatttatcca gttggtacaa acctacaatc aattatttga agaaaaccct
      601 attaacgcaa gtggagtaga tgctaaagcg attctttctg cacgattgag taaatcaaga
      661 cgattagaaa atctcattgc tcagctcccc ggtgagaaga aaaatggctt atttgggaat
      721 ctcattgctt tgtcattggg tttgacccct aattttaaat caaattttga tttggcagaa
      781 gatgctaaat tacagctttc aaaagatact tacgatgatg atttagataa tttattggcg
      841 caaattggag atcaatatgc tgatttgttt ttggcagcta agaatttatc agatgctatt
      901 ttactttcag atatcctaag agtaaatact gaaataacta aggctcccct atcagcttca
      961 atgattaaac gctacgatga acatcatcaa gacttgactc ttttaaaagc tttagttcga
     1021 caacaacttc cagaaaagta taaagaaatc ttttttgatc aatcaaaaaa cggatatgca
     1081 ggttatattg atgggggagc tagccaggaa gagttctaca agttcatcaa gcccatcctg
     1141 gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg
     1201 aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac
     1261 gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc
     1321 gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc
     1381 agattcgcct ggatgacaag aaagtccgaa gaaacgatta ctccctggaa ttttgaggaa
     1441 gttgtcgata aaggtgcgtc agctcaatcg ttcatcgaga ggatgaccgc ctttgacaag
     1501 aatttaccga acgaaaaagt attgcctaag cacagtttac tttacgagta tttcacagtg
     1561 tacaatgaac tcacgaaagt taagtatgtc actgagggca tgcgtaaacc cgcctttcta
     1621 agcggagaac agaagaaagc aatagtagat ctgttattca agaccaaccg caaagtgaca
     1681 gttaagcaat tgaaagagga ctactttaag aaaattgaat gcttcgattc tgtcgagatc
     1741 tccggggtag aagatcgatt taatgcgtca cttggtacgt atcatgacct cctaaagata
     1801 attaaagata aggacttcct ggataacgaa gagaatgaag atatcttaga agatatagtg
     1861 ttgactctta ccctctttga agatcgggaa atgattgagg aaagactaaa aacatacgct
     1921 cacctgttcg acgataaggt tatgaaacag ttaaagaggc gtcgctatac gggctgggga
     1981 gccttgtcgc ggaaacttat caacgggata agagacaagc aaagtggtaa aactattctc
     2041 gattttctaa agagcgacgg cttcgccaat aggaacttta tggccctgat ccatgatgac
     2101 tctttaacct tcaaagagga tatacaaaag gcacaggttt ccggacaagg ggactcattg
     2161 cacgaacata ttgcgaatct tgctggttcg ccagccatca aaaagggcat actccagaca
     2221 gtcaaagtag tggatgagct agttaaggtc atgggacgtc acaaaccgga aaacattgta
     2281 atcgagatgg cacgcgaaaa tcaaacgact cagaaggggc aaaaaaacag tcgagagcgg
     2341 atgaagagaa tagaagaggg tattaaagaa ctgggcagcc agatcttaaa ggagcatcct
     2401 gtggaaaata cccaattgca gaacgagaaa ctttacctct attacctaca aaatggaagg
     2461 gacatgtatg ttgatcagga actggacata aaccgtttat ctgattacga cgtcgatgcg
     2521 atcgtacccc aatccttttt gaaggacgat tcaatcgaca ataaagtgct tacacgctcg
     2581 gataagaacc gagggaaaag tgacaatgtt ccaagcgagg aagtcgtaaa gaaaatgaag
     2641 aactattggc ggcagctcct aaatgcgaaa ctgataacgc aaagaaagtt cgataactta
     2701 actaaagctg agaggggtgg cttgtctgaa cttgacaagg ccggatttat taaacgtcag
     2761 ctcgtggaaa cccgcgccat cacaaagcat gttgcgcaga tactagattc ccgaatgaat
     2821 acgaaatacg acgagaacga taagctgatt cgggaagtca aagtaatcac tttaaagtca
     2881 aaattggtgt cggacttcag aaaggatttt caattctata aagttaggga gataaataac
     2941 taccaccatg cgcacgacgc ttatcttaat gccgtcgtag ggaccgcact cattaagaaa
     3001 tacccgaagc tagaaagtga gtttgtgtat ggtgattaca aagtttatga cgtccgtaag
     3061 atgatcgcga aaagcgaaca ggagataggc aaggctacag ccaaatactt cttttattct
     3121 aacattatga atttctttaa gacggaaatc actctggcaa acggagagat acgcaaacga
     3181 cctttaattg aaaccaatgg ggagacaggt gaaatcgtat gggataaggg ccgggacttc
     3241 gcgacggtga gaaaagtttt gtccatgccc caagtcaaca tagtaaagaa aactgaggtg
     3301 cagaccggag ggttttcaaa ggaatcgatt cttccaaaaa ggaatagtga taagctcatc
     3361 gctcgtaaaa aggactggga tccaaaaaaa tatggtggtt ttgatagtcc aacggtagct
     3421 tattcagtcc tagtggttgc taaggtggaa aaagggaaat cgaagaagtt aaaatccgtt
     3481 aaagagttac tagggatcac aattatggaa agaagttcct ttgaaaaaaa tccgattgac
     3541 tttttagaag ctaaaggata taaggaagtt aaaaaagact taatcattaa actacctaaa
     3601 tatagtcttt ttgagttaga aaacggtcgt aaacggatgc tggctagtgc cggagaatta
     3661 caaaaaggaa atgagctggc tctgccaagc aaatatgtga attttttata tttagctagt
     3721 cattatgaaa agttgaaggg tagtccagaa gataacgaac aaaaacaatt gtttgtggag
     3781 cagcataagc attatttaga tgagattatt gagcaaatca gtgaattttc taagcgtgtt
     3841 attttagcag atgccaattt agataaagtt cttagtgcat ataacaaaca tagagacaaa
     3901 ccaatacgtg aacaagcaga aaatattatt catttattta cgttgacgaa tcttggagct
     3961 cccgctgctt ttaaatattt tgatacaaca attgatcgta aacgatatac gtctacaaaa
     4021 gaagttttag atgccactct tatccatcaa tccatcactg gtctttatga aacacgcatt
     4081 gatttgagtc agctaggagg tgacggttct cccaagaaga agaggaaagt ctcgagcgac
     4141 tacaaagacc atgacggtga ttataaagat catgacatcg attacaagga tgacgatgac
     4201 aaggctgcag gaggcggtgg aagcgggcgc gccgacgcgc tggacgattt cgatctcgac
     4261 atgctgggtt ctgatgccct cgatgacttt gacctggata tgttgggaag cgacgcattg
     4321 gatgactttg atctggacat gctcggctcc gatgctctgg acgatttcga tctcgatatg
     4381 ttacatcacc accaccatca ctaa
//
