LOCUS       sequence-274177-        7155 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FnCpf1 Gateway entry plasmid with N623R and K687R mutations.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7155)
  AUTHORS   Zhong Z, Zhang Y, You Q, Tang X, Ren Q, Liu S, Yang L, Wang Y, Liu 
            X, Liu B, Zhang T, Zheng X, Le Y, Zhang Y, Qi Y
  TITLE     Plant Genome Editing Using FnCpf1 and LbCpf1 Nucleases at Redefined 
            and Altered PAM Sites.
  JOURNAL   Mol Plant. 2018 Jul 2;11(7):999-1002. doi: 
            10.1016/j.molp.2018.03.008. Epub 2018 Mar 20.
  PUBMED    29567452
REFERENCE   2  (bases 1 to 7155)
  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: 
            "10.1016/j.molp.2018.03.008"; journalName: "Mol Plant"; date: 
            "2018-07-2- 2"; volume: "11"; issue: "7"; pages: "999-1002"
COMMENT     Sequence Label: pYPQ239-RR
FEATURES             Location/Qualifiers
     source          1..7155
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      18..270
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    315..438
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(455..474)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(456..474)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(479..495)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(479..495)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             926..1717
                     /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      1810..2398
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2299..2318
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2552..2569
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2728..2755)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2847..2933)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2982..3004
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2996..3013
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2997..3013
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3029..3128
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3153..3173
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7095..7142
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 ggcggccgcc gacgtccgat cgttcaaaca tttggcaata aagtttctta agattgaatc
       61 ctgttgccgg tcttgcgatg attatcatat aatttctgtt gaattacgtt aagcatgtaa
      121 taattaacat gtaatgcatg acgttattta tgagatgggt ttttatgatt agagtcccgc
      181 aattatacat ttaatacgcg atagaaaaca aaatatagcg cgcaaactag gataaattat
      241 cgcgcgcggt gtcatctatg ttactagatc gggaattgat cccccctcga cagcttccgg
      301 aaagggcgaa ttcgcaactt tgtatacaaa agttgaacga gaaacgtaaa atgatataaa
      361 tatcaatata ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca
      421 acatatccag tcactatgcc atccagctga tatcccctat agtgagtcgt attacatggt
      481 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
      541 agataaaaat atatcatcat gcctcctctg gaccagccag gacagaaatg cctcgacttc
      601 gctgctaccc aaggttgccg ggtgacgcac accgtggaaa cggatgaagg cacgaaccca
      661 gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa
      721 ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc
      781 ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg
      841 ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag
      901 tcgccctaaa acaaagttaa acattatgag ggaagcggtg atcgccgaag tatcgactca
      961 actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca
     1021 tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt
     1081 tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga
     1141 aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt
     1201 gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg
     1261 gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc
     1321 tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga
     1381 actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct
     1441 atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg
     1501 catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc
     1561 aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct
     1621 tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta
     1681 cgtgaaaggc gagatcacca aggtagtcgg caaataaccc tcgagccacc catgaccaaa
     1741 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1801 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1861 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1921 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     1981 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2041 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     2101 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     2161 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2221 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2281 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2341 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2401 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2461 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     2521 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     2581 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     2641 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
     2701 gagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
     2761 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
     2821 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
     2881 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
     2941 gcagttccct actctcgcgt taacgcttgc atggatgttt tcccagtcac gacgttgtaa
     3001 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
     3061 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
     3121 agcaggctcc gaattcgccc ttcaccatgg ctcctaagaa gaagcggaag gttggtattc
     3181 acggggtgcc tgcggcttcg atctatcaag aattcgttaa taagtattcg ctttctaaaa
     3241 cactgagatt cgaactcata cctcaaggaa agacactcga gaacataaag gctaggggcc
     3301 tcatattgga tgatgaaaag agagccaagg actataaaaa agccaaacag ataatcgaca
     3361 agtaccacca attcttcata gaagaaatcc tttccagcgt ctgcattagc gaggatttgt
     3421 tgcaaaatta cagcgatgtc tacttcaagc ttaagaagag cgatgacgac aatctccaga
     3481 aagacttcaa atcagcgaag gacacaatta agaagcagat cagcgagtat atcaaagata
     3541 gtgaaaaatt caagaacctt ttcaatcaga acctgataga cgcaaaaaaa ggacaagaaa
     3601 gcgacctcat tctttggttg aagcagtcta aggacaacgg gattgagctt tttaaggcga
     3661 atagcgatat aaccgacatc gacgaggcgc ttgagatcat aaagtcgttt aagggatgga
     3721 caacatactt caaaggcttc catgagaatc gcaaaaacgt ctactccagc aacgacattc
     3781 caacgtcgat tatatacaga attgttgatg ataacctccc taaattcctc gaaaataagg
     3841 caaaatatga aagccttaaa gataaagcgc ctgaagcaat caattatgaa caaatcaaaa
     3901 aggatcttgc tgaagaattg acgtttgata tagactacaa gacgtcagaa gttaaccaga
     3961 gggtgttttc actcgacgag gtgtttgaga ttgctaattt caacaactac ctcaatcaga
     4021 gtgggatcac gaagttcaac actattatag gtggtaagtt cgtgaatggt gagaacacta
     4081 aaagaaaagg gattaacgaa tatataaacc tttatagtca acagatcaac gacaagactt
     4141 tgaagaaata taagatgagc gtcctcttca agcagatact cagtgacacg gaatccaaga
     4201 gctttgtgat cgacaagctc gaagatgact cggatgtggt cactaccatg caatccttct
     4261 acgagcaaat tgccgctttc aaaactgtgg aggagaagag tataaaggag acactgtctc
     4321 tgctttttga cgatcttaag gcccagaaat tggatctttc caaaatatat ttcaagaatg
     4381 ataagtcact tacggacctt tcccaacaag tttttgacga ttattcagtt attggtacgg
     4441 cggttcttga gtacattacg cagcagatag cccccaagaa tctggacaac ccctctaaga
     4501 aagaacagga gttgatagcg aagaagacag agaaggcgaa atacctctcg ctggagacca
     4561 taaaattggc actggaagaa tttaacaagc atcgcgacat agataaacag tgccgcttcg
     4621 aggaaatttt ggcaaatttt gcagccattc caatgatttt cgacgagata gcgcaaaaca
     4681 aggataattt ggcacaaatt tcaataaaat atcaaaacca gggcaagaag gacctcttgc
     4741 aggcttcggc agaggatgat gttaaggcta ttaaagactt gttggaccaa acgaacaatc
     4801 tgttgcacaa gttgaaaatt ttccacatta gtcaaagcga ggataaagca aacatattgg
     4861 acaaagacga gcacttttat ctggtgtttg aggagtgcta cttcgagctt gctaatattg
     4921 tcccacttta taataaaata agaaactaca ttacgcaaaa gccatattca gatgaaaagt
     4981 ttaagctcaa tttcgaaaat agtactcttg cccgcggctg ggacaagaat aaggagccag
     5041 ataataccgc catacttttt atcaaagatg ataaatatta tcttggggtg atgaataaga
     5101 agaataataa gatcttcgat gataaagcga taaaggaaaa taagggtgaa ggctataaaa
     5161 aaattgttta caaactgttg ccgggagcaa ataaaatgct ccccagggtt tttttttcgg
     5221 caaagagcat taaattttac aatccttcag aagacattct gcgcataaga aatcattcga
     5281 cacacactaa aaatggttcg ccacaaaagg gctacgagaa atttgaattc aacattgagg
     5341 actgtcggaa gttcattgat ttctacaagc agtccatctc caagcacccg gagtggaaag
     5401 attttgggtt tcggttttcc gacacgcaga gatacaacag cattgatgaa ttttatagag
     5461 aggtcgagaa tcaaggttat aagcttacct ttgaaaacat ttctgaatca tacattgatt
     5521 cagtggtcaa tcagggcaaa ctctatcttt ttcaaatata caacaaggac tttagtgctt
     5581 atagtaaagg gcggcccaat ttgcatactc tctattggaa agcgctgttt gatgagcgga
     5641 accttcaaga cgtcgtgtat aagctcaacg gggaagccga gctcttttac cgcaagcagt
     5701 ccataccgaa aaaaataaca caccctgcca aagaagccat cgccaacaag aataaagaca
     5761 atcctaaaaa agagtccgtc ttcgaatatg atcttattaa ggacaagagg tttacagaag
     5821 ataaattttt cttccattgt cccataacta tcaatttcaa aagctctggc gcgaacaaat
     5881 ttaacgacga aatcaatctc ttgttgaaag aaaaagccaa cgatgtgcac attctgtcga
     5941 tcgacagggg agagcgccac ttggcatact acacccttgt tgatgggaaa ggaaatatta
     6001 ttaaacagga cacatttaat atcatcggca acgatcgcat gaagaccaac tatcatgaca
     6061 aactggcagc aattgaaaag gaccgcgact cagcgagaaa agactggaag aagatcaata
     6121 atatcaaaga aatgaaagag ggttatttgt ctcaagtggt ccatgagatc gcgaagttgg
     6181 tcattgaata taatgccata gtggtcttcg aagatctgaa ttttggattt aagcgcggca
     6241 ggttcaaagt cgaaaaacag gtctaccaaa agttggaaaa gatgctcatc gaaaagctga
     6301 attaccttgt cttcaaagat aacgaattcg ataaaaccgg gggggtcttg agggcctacc
     6361 aactgactgc accctttgag acttttaaaa agatgggtaa acagacagga ataatttact
     6421 atgttcctgc cggtttcact agcaagattt gccccgttac cggattcgtg aatcaactct
     6481 atcccaaata cgaatccgtg agcaagagtc aggaattctt ctccaaattt gataaaatat
     6541 gctataatct cgacaaaggt tatttcgagt tctcgttcga ctataagaac ttcggggata
     6601 aggctgccaa gggaaagtgg actatagcaa gctttggtag tcgccttata aattttagga
     6661 acagcgacaa gaatcacaac tgggacactc gggaagtcta cccaacaaaa gaactggaga
     6721 aactcttgaa ggattatagt atcgagtatg ggcatgggga gtgtatcaag gcagcgattt
     6781 gtggagagtc cgacaaaaag ttttttgcta aactcacctc ggtgctcaac actatcctcc
     6841 agatgagaaa ttcaaaaaca gggacagagc tcgattacct cattagcccc gttgccgacg
     6901 tcaatggaaa ctttttcgac tcaagacagg ctccaaaaaa catgccgcaa gatgcggacg
     6961 cgaatggggc ctatcacata ggcctgaaag ggcttatgct ccttgggaga attaaaaata
     7021 accaagaagg caaaaaactc aacctcgtca ttaagaacga agaatacttc gaatttgttc
     7081 agaacaggaa taacaagcgt cctgctgcca ccaaaaaggc cggacaggct aagaaaaaga
     7141 agtgagacga ctagt
//
