LOCUS       40924_48248        6939 bp DNA     circular SYN 13-MAY-2021
DEFINITION  LbCpf1 Gateway entry plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
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)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6939)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat
            Plants."; date: "2017-02-17"; pages: "
            10.1038/nplants.2017.18"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(562..589)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(681..767)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     831..847
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    863..962
                     /label=attL1
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             987..1007
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             1032..4712
                     /codon_start=1
                     /label=LbCpf1
                     /note="CRISPR-associated protein from Lachnospiraceae
                     bacterium ND2006 (Zetsche et al., 2015)"
                     /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
                     /label=nucleoplasmin NLS
                     /note="bipartite nuclear localization signal from
                     nucleoplasmin"
                     /translation="KRPAATKKAGQAKKKK"
     terminator      4791..5043
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    5088..5211
                     /label=attR5
                     /note="recombination site for the Gateway(R) LR reaction"
     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"
     CDS             5699..6487
                     /codon_start=1
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      6583..6939
                     /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
//
