LOCUS       sequence-147770-        5172 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expressing his-MBP tagged AsCpf1 (E.coli codon optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5172)
  AUTHORS   Hur JK, Kim K, Been KW, Baek G, Ye S, Hur JW, Ryu SM, Lee YS, Kim JS
  TITLE     Targeted mutagenesis in mice by electroporation of Cpf1 
            ribonucleoproteins.
  JOURNAL   Nat Biotechnol. 2016 Jun 6. doi: 10.1038/nbt.3596.
  PUBMED    27272385
REFERENCE   2  (bases 1 to 5172)
  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. 2016 Jun 6. doi: 10.1038/nbt.3596."
COMMENT     Sequence Label: pDEST-hisMBP-AsCpf1-EC
FEATURES             Location/Qualifiers
     source          1..5172
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             10..27
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      976..985
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1093..1116
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     protein_bind    1129..1153
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1168..1188
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1195..5115
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARND
                     HYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRN
                     AIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFD
                     KFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHF
                     ENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVL
                     NLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLL
                     RNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTG
                     KITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTL
                     KKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKA
                     RNYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYK
                     ALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEP
                     LEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSID
                     LSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKG
                     HHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLK
                     DQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFF
                     HVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQR
                     SLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAV
                     VVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQ
                     FTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYD
                     VKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIEN
                     HRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQM
                     RNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKES
                     KDLKLQNGISNQDWLAYIQELRN"
     CDS             5116..5136
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5143..5169
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             5143..5169
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 atgaaaatcc atcaccatca ccatcacgaa gaaggtaaac tggtaatctg gattaacggc
       61 gataaaggct ataacggtct cgctgaagtc ggtaagaaat tcgagaaaga taccggaatt
      121 aaagtcaccg ttgagcatcc ggataaactg gaagagaaat tcccacaggt tgcggcaact
      181 ggcgatggcc ctgacattat cttctgggca cacgaccgct ttggtggcta cgctcaatct
      241 ggcctgttgg ctgaaatcac cccggacaaa gcgttccagg acaagctgta tccgtttacc
      301 tgggatgccg tacgttacaa cggcaagctg attgcttacc cgatcgctgt tgaagcgtta
      361 tcgctgattt ataacaaaga tctgctgccg aacccgccaa aaacctggga agagatcccg
      421 gcgctggata aagaactgaa agcgaaaggt aagagcgcgc tgatgttcaa cctgcaagaa
      481 ccgtacttca cctggccgct gattgctgct gacgggggtt atgcgttcaa gtatgaaaac
      541 ggcaagtacg acattaaaga cgtgggcgtg gataacgctg gcgcgaaagc gggtctgacc
      601 ttcctggttg acctgattaa aaacaaacac atgaatgcag acaccgatta ctccatcgca
      661 gaagctgcct ttaataaagg cgaaacagcg atgaccatca acggcccgtg ggcatggtcc
      721 aacatcgaca ccagcaaagt gaattatggt gtaacggtac tgccgacctt caagggtcaa
      781 ccatccaaac cgttcgttgg cgtgctgagc gcaggtatta acgccgccag tccgaacaaa
      841 gagctggcaa aagagttcct cgaaaactat ctgctgactg atgaaggtct ggaagcggtt
      901 aataaagaca aaccgctggg tgccgtagcg ctgaagtctt acgaggaaga gttggcgaaa
      961 gatccacgta ttgccgccac catggaaaac gcccagaaag gtgaaatcat gccgaacatc
     1021 ccgcagatgt ccgctttctg gtatgccgtg cgtactgcgg tgatcaacgc cgccagcggt
     1081 cgtcagactg tcgatgaagc cctgaaagac gcgcagacta attcgatcac aagtttgtac
     1141 aaaaaagcag gcttcatggg tgctcctcca aaaaagaaga gaaaggtagg tggtatgaca
     1201 cagtttgaag gcttcaccaa tctctaccag gtcagcaaga cgctacgttt tgagcttatc
     1261 ccgcagggaa aaaccctgaa acacattcag gaacaggggt tcatagagga agataaggcg
     1321 cgtaacgacc attataaaga actgaagcct ataatcgacc gtatttataa aacgtacgcg
     1381 gatcagtgcc tgcagctggt tcagctggat tgggagaatc tgtccgcggc tattgatagc
     1441 tatcgcaaag agaagaccga ggaaacccgt aacgcactga ttgaagagca ggcgacctat
     1501 cggaatgcga tccatgatta cttcatcggc cgcaccgaca acctgaccga tgcaattaac
     1561 aaacgtcacg cagagattta caaaggtctg tttaaagcag agttattcaa tggcaaggtt
     1621 ctgaaacagc tgggtacggt caccaccacc gaacacgaaa acgcactgct gaggagcttt
     1681 gataaattta ccacatattt cagcggtttc tatgaaaatc gtaagaatgt atttagcgcc
     1741 gaagatattt ccaccgcaat tcctcatcgt attgtgcagg ataattttcc gaagtttaaa
     1801 gaaaattgtc atatttttac ccgtctgatc accgcggtac cgagcctgcg agagcatttt
     1861 gaaaacgtta agaaagccat tggaattttt gtcagtacca gcattgaaga agtgttttcg
     1921 ttcccgttct ataaccaact gctgacccag acccagattg atctgtacaa tcagctgctg
     1981 gggggcataa gccgcgaggc aggtaccgaa aagataaagg gactcaatga ggtgctgaat
     2041 ctggcaattc agaagaatga tgaaacggct catatcattg ctagcctgcc gcatcgtttc
     2101 attcccctgt ttaagcaaat cctgagcgat cgcaatacac tgagctttat cctcgaagag
     2161 tttaaatcgg acgaagaagt tatccagagc ttttgcaaat acaaaaccct gctgcggaac
     2221 gaaaatgtgc tggagaccgc tgaagcactg tttaatgaac tgaactcgat cgacctcacc
     2281 catattttta tatcccacaa aaaactggaa accataagca gcgctctgtg tgaccattgg
     2341 gataccctgc gcaacgccct gtatgaacgg cgtatcagcg agctgaccgg gaaaatcacc
     2401 aaatccgcaa aggaaaaagt tcagcgtagt ctgaaacacg aggacatcaa cctgcaagaa
     2461 attattagcg cagcaggtaa agagctgagc gaagcattca aacagaaaac cagcgaaatc
     2521 ctgagccatg cccatgctgc actggatcag ccgctgccga ccaccctgaa aaaacaggag
     2581 gaaaaggaga ttctgaaaag ccaactggac agcctgctgg gcctgtatca cctgctggac
     2641 tggtttgcag tcgatgagag caacgaggtt gatcctgagt tctccgctcg tctgaccgga
     2701 atcaagctgg agatggaacc gagtctgtcg ttttacaata aagcgcgtaa ttacgcgacc
     2761 aagaaaccgt atagcgtgga aaaattcaaa ctgaactttc agatgccgac ccttgcaagc
     2821 ggatgggacg ttaacaaaga aaaaaacaat ggggcaattc tgtttgtgaa aaatggcctc
     2881 tattatctgg gtatcatgcc gaaacagaaa gggcgctaca aagccctgtc atttgagccg
     2941 accgagaaaa cctcagaggg tttcgacaag atgtactacg attatttccc ggatgcggca
     3001 aaaatgatac ccaaatgtag cacccaactg aaggcagtta cagcccactt tcagacccat
     3061 accaccccga tcctgctgtc gaacaatttt atagagccgc tggaaattac caaagagatt
     3121 tatgatctga ataatccgga aaaggagccc aagaaatttc agacggcgta tgcaaaaaag
     3181 accggggatc agaaaggtta tcgtgaagcg ctgtgcaaat ggattgactt tacccgtgac
     3241 tttctgtcaa aatataccaa aacgacgagc attgatctga gcagcctacg tccgagcagc
     3301 caatataagg atctgggcga atattacgcc gaactgaatc cgctgctcta ccatatttcc
     3361 ttccaacgaa tcgctgaaaa agaaataatg gacgccgttg aaaccggcaa actgtatctg
     3421 tttcaaatct acaacaaaga tttcgccaaa ggccatcacg gtaagccgaa cctgcatacc
     3481 ctgtattgga ccggtctgtt tagcccggag aatctggcca aaaccagcat caagctgaac
     3541 ggacaggcag aactgtttta ccgccccaaa agccgtatga aaaggatggc acaccgcctg
     3601 ggcgaaaaaa tgctgaataa gaaactcaaa gatcagaaaa cgccgatacc ggataccctt
     3661 tatcaggagc tgtatgatta tgttaaccac cggctgagcc atgacctgag cgacgaagcg
     3721 cgtgcactgc tgccgaacgt gattaccaag gaagtctcgc atgaaattat taaagatcgg
     3781 cgcttcacca gtgataaatt tttcttccat gtaccgatca ccctgaatta tcaagccgca
     3841 aatagccctt ccaaatttaa tcaacgcgtg aatgcgtacc tgaaagagca tccggagacc
     3901 ccaattattg gcatagaccg aggagaacgc aatctcattt atatcaccgt cattgatagc
     3961 accggtaaga tcctggaaca gcgtagcctg aataccattc agcagtttga ctaccagaaa
     4021 aagctggaca acagagaaaa ggaacgtgta gccgcccggc aggcttggag tgtggtgggt
     4081 actatcaagg atctgaagca ggggtatctc tcccaagtta tccatgaaat tgtcgatcta
     4141 atgattcact atcaagcagt agtggtactg gaaaatctga atttcggttt caaaagcaaa
     4201 cgtacaggga tcgctgaaaa agccgtttat cagcagttcg agaaaatgct gatagacaag
     4261 ctgaattgcc tggttctgaa agattatccg gcagagaagg tgggcggtgt gctgaacccg
     4321 taccagctga ctgatcaatt tacgagcttt gcaaaaatgg gaacgcagag cggtttcctg
     4381 ttctatgttc cggcgccata taccagcaag atagacccgc tgacaggttt cgtagatccg
     4441 tttgtctgga aaaccattaa aaatcatgaa agtcgcaaac attttctgga gggctttgat
     4501 tttctgcact atgacgtgaa aaccggcgac ttcattctgc attttaaaat gaaccgtaat
     4561 ctgtcctttc agcgcggcct gcctggcttt atgccggcgt gggacattgt ttttgaaaag
     4621 aatgagacac agtttgatgc caaaggtacc ccctttattg cggggaaacg cattgtgccc
     4681 gttatagaaa atcaccgctt caccggacgg tatagggact tgtacccggc aaatgaattg
     4741 atagcgctgc tggaggagaa aggtattgtc tttcgggatg gatcaaacat cctgccgaag
     4801 ctgctggaga acgatgacag ccacgcaata gacaccatgg tagcgctgat ccgaagcgtg
     4861 ctgcagatgc gtaacagtaa tgcggctacg ggggaagact acattaatag cccggtccgt
     4921 gatctgaacg gcgtttgttt cgatagcaga tttcaaaatc cggagtggcc gatggatgcc
     4981 gatgccaatg gagcttacca tatcgctctc aaaggtcagc tcctactgaa ccatttgaaa
     5041 gaatcaaaag atctgaaact gcagaacggc atctcgaatc aggactggct ggcctacatt
     5101 caagaactga gaaacccaaa gaaaaagaga aaagtaggtg gttaccccta cgacgtgccc
     5161 gactacgcct aa
//
