LOCUS Exported 6939 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION LbCpf1 Gateway entry plasmid. ACCESSION . VERSION . KEYWORDS pYPQ230 (LbCpf1) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6939) AUTHORS Tang X, Lowder LG, Zhang T, Malzahn AA, Zheng X, Voytas DF, Zhong Z, Chen Y, Ren Q, Li Q, Kirkland ER, Zhang Y, Qi Y TITLE A CRISPR-Cpf1 system for efficient genome editing and transcriptional repression in plants. JOURNAL Nat Plants. 2017 Feb 17;3:17018. doi: 10.1038/nplants.2017.18. PUBMED 28211909 REFERENCE 2 (bases 1 to 6939) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6939 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 133..152 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 386..403 /label=L4440 /note="L4440 vector, forward primer" terminator 562..589 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 681..767 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 816..838 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 830..847 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 831..847 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 863..962 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 987..1007 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1032..4712 /codon_start=1 /product="CRISPR-associated protein from Lachnospiraceae bacterium ND2006 (Zetsche et al., 2015)" /label=LbCpf1 /translation="SKLEKFTNCYSLSKTLRFKAIPVGKTQENIDNKRLLVEDEKRAED YKGVKKLLDRYYLSFINDVLHSIKLKNLNNYISLFRKKTRTEKENKELENLEINLRKEI AKAFKGNEGYKSLFKKDIIETILPEFLDDKDEIALVNSFNGFTTAFTGFFDNRENMFSE EAKSTSIAFRCINENLTRYISNMDIFEKVDAIFDKHEVQEIKEKILNSDYDVEDFFEGE FFNFVLTQEGIDVYNAIIGGFVTESGEKIKGLNEYINLYNQKTKQKLPKFKPLYKQVLS DRESLSFYGEGYTSDEEVLEVFRNTLNKNSEIFSSIKKLEKLFKNFDEYSSAGIFVKNG PAISTISKDIFGEWNVIRDKWNAEYDDIHLKKKAVVTEKYEDDRRKSFKKIGSFSLEQL QEYADADLSVVEKLKEIIIQKVDEIYKVYGSSEKLFDADFVLEKSLKKNDAVVAIMKDL LDSVKSFENYIKAFFGEGKETNRDESFYGDFVLAYDILLKVDHIYDAIRNYVTQKPYSK DKFKLYFQNPQFMGGWDKDKETDYRATILRYGSKYYLAIMDKKYAKCLQKIDKDDVNGN YEKINYKLLPGPNKMLPKVFFSKKWMAYYNPSEDIQKIYKNGTFKKGDMFNLNDCHKLI DFFKDSISRYPKWSNAYDFNFSETEKYKDIAGFYREVEEQGYKVSFESASKKEVDKLVE EGKLYMFQIYNKDFSDKSHGTPNLHTMYFKLLFDENNHGQIRLSGGAELFMRRASLKKE ELVVHPANSPIANKNPDNPKKTTTLSYDVYKDKRFSEDQYELHIPIAINKCPKNIFKIN TEVRVLLKHDDNPYVIGIDRGERNLLYIVVVDGKGNIVEQYSLNEIINNFNGIRIKTDY HSLLDKKEKERFEARQNWTSIENIKELKAGYISQVVHKICELVEKYDAVIALEDLNSGF KNSRVKVEKQVYQKFEKMLIDKLNYMVDKKSNPCATGGALKGYQITNKFESFKSMSTQN GFIFYIPAWLTSKIDPSTGFVNLLKTKYTSIADSKKFISSFDRIMYVPEEDLFEFALDY KNFSRTDADYIKKWKLYSYGNRIRIFRNPKKNNVFDWEEVCLTSAYKELFNKYGINYQQ GDIRALLCEQSDKAFYSSFMALMSLMLQMRNSITGRTDVDFLISPVKNSDGIFYDSRNY EAQENAILPKNADANGAYNIARKVLWAIGQFKKAEDEKLDKVKIAISNKEWLEYAQTSV KH" CDS 4713..4760 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" terminator 4791..5043 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" protein_bind 5088..5211 /gene="mutant version of attR" /label=attR5 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(5228..5247) /label=T7 /note="T7 promoter, forward primer" promoter complement(5229..5247) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5252..5268) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5252..5268) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 5699..6490 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" rep_origin 6583..232 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga 61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag 121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg 181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag 241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc 301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc 361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc 421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag 481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgcgagcca 541 ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga 601 tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg 661 ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag 721 ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt 781 ccctactctc gcgttaacgc ttgcatggat gttttcccag tcacgacgtt gtaaaacgac 841 ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag tgacctgttc 901 gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca aaaaagcagg 961 ctccgaattc gcccttcacc atggctccta agaagaagcg gaaggttggt attcacgggg 1021 tgcctgcggc ttcaaagctc gagaaattca ccaactgtta ttcgttgagc aaaacactgc 1081 ggtttaaagc gattccagtc ggcaagactc aagagaatat agacaataag cggctgttgg 1141 tggaagatga aaagcgcgcg gaagactaca aaggggtgaa gaagttgttg gacagatact 1201 acctctcttt tatcaatgat gtcttgcact caatcaaatt gaagaatctg aacaactaca 1261 tctccctctt cagaaagaaa acaaggacag aaaaggagaa taaggaactt gaaaatttgg 1321 agatcaatct gaggaaagag atcgcgaaag cctttaaagg caacgaagga tacaaaagtc 1381 tgttcaagaa ggatataatt gagacaattt tgccagagtt cctcgatgac aaggacgaga 1441 ttgcgctggt caattcgttc aacggattca caacagcatt cacaggcttc tttgataatc 1501 gggaaaatat gttctctgag gaggcaaagt ccacttctat tgcgttcagg tgtatcaatg 1561 agaatctcac taggtacatt tccaacatgg atatctttga gaaggttgac gcaatttttg 1621 acaagcacga agttcaggag attaaggaga agatcctcaa ttccgattat gacgttgagg 1681 acttcttcga aggtgagttt tttaatttcg tgctcactca agagggtatc gacgtgtata 1741 atgcgatcat cggtgggttc gtgactgagt ccggtgaaaa gattaaggga ttgaacgagt 1801 atatcaacct ttacaaccaa aagacgaaac agaagctgcc aaagttcaag cctctttaca 1861 aacaggttct ttcagaccgc gagtcactct cgttctatgg ggagggctac acttcggatg 1921 aggaagtcct ggaggtgttc aggaatactc tcaataagaa ttcggagatt ttctcttcta 1981 taaaaaaact ggaaaagttg tttaagaatt ttgacgaata ctctagcgcc ggcatatttg 2041 tgaaaaacgg cccggccata tcaacgataa gtaaagatat cttcggcgaa tggaacgtga 2101 tcagagacaa atggaacgcg gagtatgacg atattcacct gaagaagaag gctgtcgtaa 2161 cggagaagta cgaggatgat cgcaggaaaa gcttcaaaaa gatcggaagt ttcagcctgg 2221 aacagttgca ggagtatgct gacgccgatc ttagcgtcgt cgagaagttg aaggagataa 2281 tcatccaaaa ggtcgacgag atatataaag tctatggatc aagtgaaaaa ctgttcgacg 2341 ccgacttcgt tttggagaag tccctgaaga agaacgacgc tgttgttgcc attatgaagg 2401 atctgctcga cagcgtgaag agtttcgaga actatattaa ggcttttttc ggggagggga 2461 aggagactaa cagagatgag tccttctacg gagacttcgt cctcgcgtac gatatactcc 2521 ttaaggtaga ccacatctac gacgcaatca gaaattacgt gacacaaaag ccgtacagca 2581 aggacaagtt caaactctac ttccagaacc cccagttcat gggcggctgg gacaaggaca 2641 aggaaacgga ttacagggct acgatcctga ggtatggttc aaaatactac ttggcgatta 2701 tggacaagaa gtacgccaag tgtctccaga agattgacaa agacgatgtc aatggcaatt 2761 atgagaagat caactacaag ctgcttccgg gtccgaacaa gatgctccca aaggttttct 2821 tcagcaagaa atggatggcc tactataacc caagcgagga catccagaag atttataaga 2881 acggtacgtt caagaagggc gacatgttca atcttaacga ctgtcacaag ctgatcgact 2941 tcttcaaaga ctcaattagc cggtacccaa agtggtctaa cgcctatgac ttcaactttt 3001 cggaaaccga gaagtacaag gatatagccg gattttatag agaggtggaa gagcagggct 3061 acaaggtgtc attcgagtcc gccagcaaga aggaagtgga caagctcgtg gaagagggta 3121 agctctacat gttccagatt tataataaag actttagcga taagagccac gggacaccta 3181 atctccacac aatgtatttc aagctgctct tcgacgagaa taaccacggc caaatcaggt 3241 tgtcaggagg ggctgaactc ttcatgcggc gcgctagcct taagaaggag gagcttgtag 3301 tccaccctgc gaatagtcca attgcgaata agaacccgga caatcctaaa aagactacaa 3361 cattgagcta cgacgtgtac aaggataaga ggttttccga ggatcagtac gagctccaca 3421 tcccgattgc gatcaacaag tgcccaaaga atattttcaa gataaacaca gaggtgcgtg 3481 tactcctgaa gcatgacgac aatccttacg tcattgggat tgatcggggc gagaggaacc 3541 tcctctatat tgtggtggtg gacgggaagg ggaacatagt cgaacagtac tcccttaacg 3601 aaataattaa caatttcaac ggcatccgta tcaagaccga ctaccattcg ttgctggaca 3661 agaaggagaa ggagagattt gaggcgcggc aaaattggac aagtatcgag aacatcaagg 3721 aactcaaagc aggttatatc tctcaagttg tgcataagat atgcgagctg gttgagaagt 3781 atgacgcagt gatcgctctt gaggacctca actcgggctt taagaattct agagttaaag 3841 tggagaagca ggtctatcaa aagttcgaga agatgcttat agataagctc aactacatgg 3901 tcgataagaa atcgaaccca tgtgccaccg gcggcgcact caaaggttac caaataacaa 3961 acaaattcga gtccttcaaa tcgatgagta ctcagaatgg gttcatattt tatataccgg 4021 cgtggcttac gtctaagatc gacccgtcaa ctggttttgt caacctgttg aagacgaaat 4081 acacgtccat tgccgattcg aaaaagttca tatctagttt tgatcgtatt atgtacgtcc 4141 cagaggaaga tcttttcgag tttgctctcg actacaaaaa cttttcgcgg accgatgcgg 4201 attacattaa aaaatggaaa ctctattcgt acggcaacag aatcaggatt tttcgcaacc 4261 ctaagaagaa taacgtcttt gattgggagg aagtttgctt gactagcgcg tacaaggagc 4321 tctttaataa gtatggcatt aactaccaac agggtgatat cagagcactg ctttgcgaac 4381 aatctgacaa ggctttctac tcatccttca tggctttgat gagcctgatg ctccagatga 4441 gaaattcaat tacaggcaga accgacgtgg atttcttgat ctccccggtt aaaaattctg 4501 atggcatctt ttacgatagc aggaactatg aagcgcaaga gaatgcgatt ctgccaaaaa 4561 atgcagacgc caacggtgcc tataacatcg ccaggaaagt cctgtgggcg atcggccagt 4621 tcaaaaaggc cgaagacgaa aaattggaca aggtcaaaat cgctatcagc aacaaagagt 4681 ggctggagta tgctcagaca tccgtaaagc ataagcgtcc tgctgccacc aaaaaggccg 4741 gacaggctaa gaaaaagaag tgagacgact agtggcggcc gccgacgtcc gatcgttcaa 4801 acatttggca ataaagtttc ttaagattga atcctgttgc cggtcttgcg atgattatca 4861 tataatttct gttgaattac gttaagcatg taataattaa catgtaatgc atgacgttat 4921 ttatgagatg ggtttttatg attagagtcc cgcaattata catttaatac gcgatagaaa 4981 acaaaatata gcgcgcaaac taggataaat tatcgcgcgc ggtgtcatct atgttactag 5041 atcgggaatt gatcccccct cgacagcttc cggaaagggc gaattcgcaa ctttgtatac 5101 aaaagttgaa cgagaaacgt aaaatgatat aaatatcaat atattaaatt agattttgca 5161 taaaaaacag actacataat actgtaaaac acaacatatc cagtcactat gccatccagc 5221 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc 5281 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgcctcct 5341 ctggaccagc caggacagaa atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg 5401 cacaccgtgg aaacggatga aggcacgaac ccagtggaca taagcctgtt cggttcgtaa 5461 gctgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag 5521 cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttggggtaca 5581 gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt 5641 atggagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacattat 5701 gagggaagcg gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga 5761 gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg 5821 cctgaagcca cacagtgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac 5881 aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga 5941 gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta 6001 tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat 6061 cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca 6121 tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga 6181 tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg 6241 cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa 6301 aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca 6361 gcccgtcata cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc 6421 gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt 6481 cggcaaataa ccctcgagcc acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt 6541 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct 6601 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc 6661 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc 6721 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc 6781 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc 6841 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg 6901 aacggggggt tcgtgcacac agcccagctt ggagcgaac //