LOCUS       sequence-274167-        7155 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FnCpf1 Gateway entry plasmid with N623R, K629R and N633R 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-RVR
FEATURES             Location/Qualifiers
     source          1..7155
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             157..177
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4099..4146
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      4177..4429
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    4474..4597
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4614..4633)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4615..4633)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4638..4654)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4638..4654)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5085..5876
                     /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      5969..6557
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6458..6477
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6711..6728
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6887..6914)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(7006..7092)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(7141..7155,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(7155,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagcagg ctccgaattc gcccttcacc atggctccta agaagaagcg gaaggttggt
      181 attcacgggg tgcctgcggc ttcgatctat caagaattcg ttaataagta ttcgctttct
      241 aaaacactga gattcgaact catacctcaa ggaaagacac tcgagaacat aaaggctagg
      301 ggcctcatat tggatgatga aaagagagcc aaggactata aaaaagccaa acagataatc
      361 gacaagtacc accaattctt catagaagaa atcctttcca gcgtctgcat tagcgaggat
      421 ttgttgcaaa attacagcga tgtctacttc aagcttaaga agagcgatga cgacaatctc
      481 cagaaagact tcaaatcagc gaaggacaca attaagaagc agatcagcga gtatatcaaa
      541 gatagtgaaa aattcaagaa ccttttcaat cagaacctga tagacgcaaa aaaaggacaa
      601 gaaagcgacc tcattctttg gttgaagcag tctaaggaca acgggattga gctttttaag
      661 gcgaatagcg atataaccga catcgacgag gcgcttgaga tcataaagtc gtttaaggga
      721 tggacaacat acttcaaagg cttccatgag aatcgcaaaa acgtctactc cagcaacgac
      781 attccaacgt cgattatata cagaattgtt gatgataacc tccctaaatt cctcgaaaat
      841 aaggcaaaat atgaaagcct taaagataaa gcgcctgaag caatcaatta tgaacaaatc
      901 aaaaaggatc ttgctgaaga attgacgttt gatatagact acaagacgtc agaagttaac
      961 cagagggtgt tttcactcga cgaggtgttt gagattgcta atttcaacaa ctacctcaat
     1021 cagagtggga tcacgaagtt caacactatt ataggtggta agttcgtgaa tggtgagaac
     1081 actaaaagaa aagggattaa cgaatatata aacctttata gtcaacagat caacgacaag
     1141 actttgaaga aatataagat gagcgtcctc ttcaagcaga tactcagtga cacggaatcc
     1201 aagagctttg tgatcgacaa gctcgaagat gactcggatg tggtcactac catgcaatcc
     1261 ttctacgagc aaattgccgc tttcaaaact gtggaggaga agagtataaa ggagacactg
     1321 tctctgcttt ttgacgatct taaggcccag aaattggatc tttccaaaat atatttcaag
     1381 aatgataagt cacttacgga cctttcccaa caagtttttg acgattattc agttattggt
     1441 acggcggttc ttgagtacat tacgcagcag atagccccca agaatctgga caacccctct
     1501 aagaaagaac aggagttgat agcgaagaag acagagaagg cgaaatacct ctcgctggag
     1561 accataaaat tggcactgga agaatttaac aagcatcgcg acatagataa acagtgccgc
     1621 ttcgaggaaa ttttggcaaa ttttgcagcc attccaatga ttttcgacga gatagcgcaa
     1681 aacaaggata atttggcaca aatttcaata aaatatcaaa accagggcaa gaaggacctc
     1741 ttgcaggctt cggcagagga tgatgttaag gctattaaag acttgttgga ccaaacgaac
     1801 aatctgttgc acaagttgaa aattttccac attagtcaaa gcgaggataa agcaaacata
     1861 ttggacaaag acgagcactt ttatctggtg tttgaggagt gctacttcga gcttgctaat
     1921 attgtcccac tttataataa aataagaaac tacattacgc aaaagccata ttcagatgaa
     1981 aagtttaagc tcaatttcga aaatagtact cttgcccgcg gctgggacaa gaatgtggag
     2041 ccagatagga ccgccatact ttttatcaaa gatgataaat attatcttgg ggtgatgaat
     2101 aagaagaata ataagatctt cgatgataaa gcgataaagg aaaataaggg tgaaggctat
     2161 aaaaaaattg tttacaaact gttgccggga gcaaataaaa tgctccccaa ggtttttttt
     2221 tcggcaaaga gcattaaatt ttacaatcct tcagaagaca ttctgcgcat aagaaatcat
     2281 tcgacacaca ctaaaaatgg ttcgccacaa aagggctacg agaaatttga attcaacatt
     2341 gaggactgtc ggaagttcat tgatttctac aagcagtcca tctccaagca cccggagtgg
     2401 aaagattttg ggtttcggtt ttccgacacg cagagataca acagcattga tgaattttat
     2461 agagaggtcg agaatcaagg ttataagctt acctttgaaa acatttctga atcatacatt
     2521 gattcagtgg tcaatcaggg caaactctat ctttttcaaa tatacaacaa ggactttagt
     2581 gcttatagta aagggcggcc caatttgcat actctctatt ggaaagcgct gtttgatgag
     2641 cggaaccttc aagacgtcgt gtataagctc aacggggaag ccgagctctt ttaccgcaag
     2701 cagtccatac cgaaaaaaat aacacaccct gccaaagaag ccatcgccaa caagaataaa
     2761 gacaatccta aaaaagagtc cgtcttcgaa tatgatctta ttaaggacaa gaggtttaca
     2821 gaagataaat ttttcttcca ttgtcccata actatcaatt tcaaaagctc tggcgcgaac
     2881 aaatttaacg acgaaatcaa tctcttgttg aaagaaaaag ccaacgatgt gcacattctg
     2941 tcgatcgaca ggggagagcg ccacttggca tactacaccc ttgttgatgg gaaaggaaat
     3001 attattaaac aggacacatt taatatcatc ggcaacgatc gcatgaagac caactatcat
     3061 gacaaactgg cagcaattga aaaggaccgc gactcagcga gaaaagactg gaagaagatc
     3121 aataatatca aagaaatgaa agagggttat ttgtctcaag tggtccatga gatcgcgaag
     3181 ttggtcattg aatataatgc catagtggtc ttcgaagatc tgaattttgg atttaagcgc
     3241 ggcaggttca aagtcgaaaa acaggtctac caaaagttgg aaaagatgct catcgaaaag
     3301 ctgaattacc ttgtcttcaa agataacgaa ttcgataaaa ccgggggggt cttgagggcc
     3361 taccaactga ctgcaccctt tgagactttt aaaaagatgg gtaaacagac aggaataatt
     3421 tactatgttc ctgccggttt cactagcaag atttgccccg ttaccggatt cgtgaatcaa
     3481 ctctatccca aatacgaatc cgtgagcaag agtcaggaat tcttctccaa atttgataaa
     3541 atatgctata atctcgacaa aggttatttc gagttctcgt tcgactataa gaacttcggg
     3601 gataaggctg ccaagggaaa gtggactata gcaagctttg gtagtcgcct tataaatttt
     3661 aggaacagcg acaagaatca caactgggac actcgggaag tctacccaac aaaagaactg
     3721 gagaaactct tgaaggatta tagtatcgag tatgggcatg gggagtgtat caaggcagcg
     3781 atttgtggag agtccgacaa aaagtttttt gctaaactca cctcggtgct caacactatc
     3841 ctccagatga gaaattcaaa aacagggaca gagctcgatt acctcattag ccccgttgcc
     3901 gacgtcaatg gaaacttttt cgactcaaga caggctccaa aaaacatgcc gcaagatgcg
     3961 gacgcgaatg gggcctatca cataggcctg aaagggctta tgctccttgg gagaattaaa
     4021 aataaccaag aaggcaaaaa actcaacctc gtcattaaga acgaagaata cttcgaattt
     4081 gttcagaaca ggaataacaa gcgtcctgct gccaccaaaa aggccggaca ggctaagaaa
     4141 aagaagtgag acgactagtg gcggccgccg acgtccgatc gttcaaacat ttggcaataa
     4201 agtttcttaa gattgaatcc tgttgccggt cttgcgatga ttatcatata atttctgttg
     4261 aattacgtta agcatgtaat aattaacatg taatgcatga cgttatttat gagatgggtt
     4321 tttatgatta gagtcccgca attatacatt taatacgcga tagaaaacaa aatatagcgc
     4381 gcaaactagg ataaattatc gcgcgcggtg tcatctatgt tactagatcg ggaattgatc
     4441 ccccctcgac agcttccgga aagggcgaat tcgcaacttt gtatacaaaa gttgaacgag
     4501 aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta
     4561 cataatactg taaaacacaa catatccagt cactatgcca tccagctgat atcccctata
     4621 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc
     4681 tctgatgtta cattgcacaa gataaaaata tatcatcatg cctcctctgg accagccagg
     4741 acagaaatgc ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac
     4801 ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt
     4861 agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg
     4921 cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg
     4981 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg
     5041 atgttacgca gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga
     5101 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac
     5161 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca
     5221 gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt
     5281 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg
     5341 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg
     5401 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca
     5461 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg
     5521 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc
     5581 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg
     5641 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg
     5701 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg
     5761 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
     5821 tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct
     5881 cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
     5941 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6001 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6061 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6121 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6181 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6241 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6301 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6361 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6421 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6481 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6541 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6601 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6661 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     6721 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     6781 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     6841 caattaatac gcgtaccgcg agccaggaag agtttgtaga aacgcaaaaa ggccatccgt
     6901 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca
     6961 ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag
     7021 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt
     7081 tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgcttgca tggatgtttt
     7141 cccagtcacg acgtt
//
