LOCUS       sequence-300002-        3921 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Lentiviral vector expressing human codon-optimized _C-terminal 
            Myc-tagged _Acidaminococcus sp. BV3L6 (As)-Cas12a nuclease with N- 
            and C-terminal NLS and Neomycin/G418/Geneticin resistance from 
            EF-1alpha promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3921)
  AUTHORS   Gonatopoulos-Pournatzis T, Aregger M, Brown KR, Farhangmehr S, 
            Braunschweig U, Ward HN, Ha KCH, Weiss A, Billmann M, Durbic T, 
            Myers CL, Blencowe BJ, Moffat J
  TITLE     Genetic interaction mapping and exon-resolution functional genomics 
            with a hybrid Cas9-Cas12a platform.
  JOURNAL   Nat Biotechnol. 2020 May;38(5):638-648. doi: 
            10.1038/s41587-020-0437-z. Epub 2020 Mar 16.
  PUBMED    32249828
REFERENCE   2  (bases 1 to 3921)
  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"; doi: 
            "10.1038/s41587-020-0437-z"; journalName: "Nat Biotechnol"; date: 
            "2020-05"; volume: "38"; issue: "5"; pages: "638-648"
COMMENT     Sequence Label: plenti-As-Cas12a-2xNLS
FEATURES             Location/Qualifiers
     source          1..3921
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1..3921
                     /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"
     regulatory      3641..3650
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atgacacagt tcgagggctt taccaacctg tatcaggtga gcaagacact gcggtttgag
       61 ctgatcccac agggcaagac cctgaagcac atccaggagc agggcttcat cgaggaggac
      121 aaggcccgca atgatcacta caaggagctg aagcccatca tcgatcggat ctacaagacc
      181 tatgccgacc agtgcctgca gctggtgcag ctggattggg agaacctgag cgccgccatc
      241 gactcctata gaaaggagaa aaccgaggag acaaggaacg ccctgatcga ggagcaggcc
      301 acatatcgca atgccatcca cgactacttc atcggccgga cagacaacct gaccgatgcc
      361 atcaataaga gacacgccga gatctacaag ggcctgttca aggccgagct gtttaatggc
      421 aaggtgctga agcagctggg caccgtgacc acaaccgagc acgagaacgc cctgctgcgg
      481 agcttcgaca agtttacaac ctacttctcc ggcttttatg agaacaggaa gaacgtgttc
      541 agcgccgagg atatcagcac agccatccca caccgcatcg tgcaggacaa cttccccaag
      601 tttaaggaga attgtcacat cttcacacgc ctgatcaccg ccgtgcccag cctgcgggag
      661 cactttgaga acgtgaagaa ggccatcggc atcttcgtga gcacctccat cgaggaggtg
      721 ttttccttcc ctttttataa ccagctgctg acacagaccc agatcgacct gtataaccag
      781 ctgctgggag gaatctctcg ggaggcaggc accgagaaga tcaagggcct gaacgaggtg
      841 ctgaatctgg ccatccagaa gaatgatgag acagcccaca tcatcgcctc cctgccacac
      901 agattcatcc ccctgtttaa gcagatcctg tccgatagga acaccctgtc tttcatcctg
      961 gaggagttta agagcgacga ggaagtgatc cagtccttct gcaagtacaa gacactgctg
     1021 agaaacgaga acgtgctgga gacagccgag gccctgttta acgagctgaa cagcatcgac
     1081 ctgacacaca tcttcatcag ccacaagaag ctggagacaa tcagcagcgc cctgtgcgac
     1141 cactgggata cactgaggaa tgccctgtat gagcggagaa tctccgagct gacaggcaag
     1201 atcaccaagt ctgccaagga gaaggtgcag cgcagcctga agcacgagga tatcaacctg
     1261 caggagatca tctctgccgc aggcaaggag ctgagcgagg ccttcaagca gaaaaccagc
     1321 gagatcctgt cccacgcaca cgccgccctg gatcagccac tgcctacaac cctgaagaag
     1381 caggaggaga aggagatcct gaagtctcag ctggacagcc tgctgggcct gtaccacctg
     1441 ctggactggt ttgccgtgga tgagtccaac gaggtggacc ccgagttctc tgcccggctg
     1501 accggcatca agctggagat ggagccttct ctgagcttct acaacaaggc cagaaattat
     1561 gccaccaaga agccctactc cgtggagaag ttcaagctga actttcagat gcctacactg
     1621 gcctctggct gggacgtgaa taaggagaag aacaatggcg ccatcctgtt tgtgaagaac
     1681 ggcctgtact atctgggcat catgccaaag cagaagggca ggtataaggc cctgagcttc
     1741 gagcccacag agaaaaccag cgagggcttt gataagatgt actatgacta cttccctgat
     1801 gccgccaaga tgatcccaaa gtgcagcacc cagctgaagg ccgtgacagc ccactttcag
     1861 acccacacaa cccccatcct gctgtccaac aatttcatcg agcctctgga gatcacaaag
     1921 gagatctacg acctgaacaa tcctgagaag gagccaaaga agtttcagac agcctacgcc
     1981 aagaaaaccg gcgaccagaa gggctacaga gaggccctgt gcaagtggat cgacttcaca
     2041 agggattttc tgtccaagta taccaagaca acctctatcg atctgtctag cctgcggcca
     2101 tcctctcagt ataaggacct gggcgagtac tatgccgagc tgaatcccct gctgtaccac
     2161 atcagcttcc agagaatcgc cgagaaggag atcatggatg ccgtggagac aggcaagctg
     2221 tacctgttcc agatctataa caaggacttt gccaagggcc accacggcaa gcctaatctg
     2281 cacacactgt attggaccgg cctgttttct ccagagaacc tggccaagac aagcatcaag
     2341 ctgaatggcc aggccgagct gttctaccgc cctaagtcca ggatgaagag gatggcacac
     2401 cggctgggag agaagatgct gaacaagaag ctgaaggatc agaaaacccc aatccccgac
     2461 accctgtacc aggagctgta cgactatgtg aatcacagac tgtcccacga cctgtctgat
     2521 gaggccaggg ccctgctgcc caacgtgatc accaaggagg tgtctcacga gatcatcaag
     2581 gataggcgct ttaccagcga caagttcttt ttccacgtgc ctatcacact gaactatcag
     2641 gccgccaatt ccccatctaa gttcaaccag agggtgaatg cctacctgaa ggagcacccc
     2701 gagacaccta tcatcggcat cgatcggggc gagagaaacc tgatctatat cacagtgatc
     2761 gactccaccg gcaagatcct ggagcagcgg agcctgaaca ccatccagca gtttgattac
     2821 cagaagaagc tggacaacag ggagaaggag agggtggcag caaggcaggc ctggtctgtg
     2881 gtgggcacaa tcaaggatct gaagcagggc tatctgagcc aggtcatcca cgagatcgtg
     2941 gacctgatga tccactacca ggccgtggtg gtgctggaga acctgaattt cggctttaag
     3001 agcaagagga ccggcatcgc cgagaaggcc gtgtaccagc agttcgagaa gatgctgatc
     3061 gataagctga attgcctggt gctgaaggac tatccagcag agaaagtggg aggcgtgctg
     3121 aacccatacc agctgacaga ccagttcacc tcctttgcca agatgggcac ccagtctggc
     3181 ttcctgtttt acgtgcctgc cccatataca tctaagatcg atcccctgac cggcttcgtg
     3241 gaccccttcg tgtggaaaac catcaagaat cacgagagcc gcaagcactt cctggagggc
     3301 ttcgactttc tgcactacga cgtgaaaacc ggcgacttca tcctgcactt taagatgaac
     3361 agaaatctgt ccttccagag gggcctgccc ggctttatgc ctgcatggga tatcgtgttc
     3421 gagaagaacg agacacagtt tgacgccaag ggcacccctt tcatcgccgg caagagaatc
     3481 gtgccagtga tcgagaatca cagattcacc ggcagatacc gggacctgta tcctgccaac
     3541 gagctgatcg ccctgctgga ggagaagggc atcgtgttca gggatggctc caacatcctg
     3601 ccaaagctgc tggagaatga cgattctcac gccatcgaca ccatggtggc cctgatccgc
     3661 agcgtgctgc agatgcggaa ctccaatgcc gccacaggcg aggactatat caacagcccc
     3721 gtgcgcgatc tgaatggcgt gtgcttcgac tcccggtttc agaacccaga gtggcccatg
     3781 gacgccgatg ccaatggcgc ctaccacatc gccctgaagg gccagctgct gctgaatcac
     3841 ctgaaggaga gcaaggatct gaagctgcag aacggcatct ccaatcagga ctggctggcc
     3901 tacatccagg agctgcgcaa c
//
