LOCUS       sequence-274170-        4989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FnCpf1 Gateway crRNA entry plasmid using Zea mays Ubi promoter and 
            double ribozyme processing.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4989)
  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 4989)
  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: pYPQ141-ZmUbi-RZ-Fn
FEATURES             Location/Qualifiers
     source          1..4989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1192..1287)
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1302..1319)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1302..1318)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1311..1333)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      1382..1468
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1560..1587
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1746..1763)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1917..2505)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1997..2016)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2598..3389)
                     /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"
     primer_bind     3820..3836
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3820..3836
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3841..3859
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        3876..3894
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3912..4011
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     ncRNA           complement(4031..4098)
                     /product="hepatitis delta virus self-cleaving ribozyme 
                     (Ferre-D'Amare and Scott, 2010)"
                     /label=HDV ribozyme
                     /ncRNA_class="ribozyme"
     ncRNA           complement(4151..4187)
                     /product="self-cleaving hammerhead ribozyme (Ferre-D'Amare 
                     and Scott, 2010)"
                     /label=hammerhead ribozyme
                     /ncRNA_class="hammerhead_ribozyme"
ORIGIN
        1 agccatccca ggattcccca aagagaaaca ctggcaagtt agcaatcaga acgtgtctga
       61 cgtacaggtc gcatccgtgt acgaacgcta gcagcacgga tctaacacaa acacggatct
      121 aacacaaaca tgaacagaag tagaactacc gggccctaac catggaccgg aacgccgatc
      181 tagagaaggt agagaggggg gggggggagg acgagcggcg taccttgaag cggaggtgcc
      241 gacgggtgga tttgggggag atctggttgt gtgtgtgtgc gctccgaaca acacgaggtt
      301 ggggaaagag ggtgtggagg gggtgtctat ttattacggc gggcgaggaa gggaaagcga
      361 aggagcggtg ggaaaggaat cccccgtagc tgccggtgcc gtgagaggag gaggaggccg
      421 cctgccgtgc cggctcacgt ctgccgctcc gccacgcaat ttctggatgc cgacagcgga
      481 gcaagtccaa cggtggagcg gaactctcga gaggggtcca gaggcagcga cagagatgcc
      541 gtgccgtctg cttcgcttgg cccgacgcga cgctgctggt tcgctggttg gtgtccgtta
      601 gactcgtcga cggcgtttaa caggctggca ttatctactc gaaacaagaa aaatgtttcc
      661 ttagtttttt taatttctta aagggtattt gtttaatttt tagtcacttt attttattct
      721 attttatatc taaattatta aataaaaaaa ctaaaataga gttttagttt tcttaattta
      781 gaggctaaaa tagaataaaa tagatgtact aaaaaaatta gtctataaaa accattaacc
      841 ctaaacccta aatggatgta ctaataaaat ggatgaagta ttatataggt gaagctattt
      901 gcaaaaaaaa aggagaacac atgcacacta aaaagataaa actgtagagt cctgttgtca
      961 aaatactcaa ttgtccttta gaccatgtct aactgttcat ttatatgatt ctctaaaaca
     1021 ctgatattat tgtagtacta tagattatat tattcgtaga gtaaagttta aatatatgta
     1081 taaagataga taaactgcac ttcaaacaag tgtgacaaaa aaaatatgtg gtaatttttt
     1141 ataacttaga catgcaatgc tcattatctc tctagaggga acgccatggg gcaacttttg
     1201 tatacaaagt tggcattata aaaaagcatt gctcatcaat ttgttgcaac gaacaggtca
     1261 ctatcagtca aaataaaatc attattgggc ccgagcttaa gactggccgt cgttttacaa
     1321 cgtcgtgact gggaaaacat ccatgcaagc gttaacgcga gagtagggaa ctgccaggca
     1381 tcaaataaaa cgaaaggctc agtcggaaga ctgggccttt cgttttatct gttgtttgtc
     1441 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgtgaagca
     1501 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaactaagca
     1561 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttcctg gctcgcggta
     1621 cgcgtattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     1681 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     1741 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     1801 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     1861 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1921 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1981 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2041 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2101 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2161 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2221 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2281 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2341 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2401 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2461 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     2521 ttttctacgg ggtctgacgc tcagtggaac gacgcgtaac tcacgttaag ggattttggt
     2581 catgggtggc tcgagggtta tttgccgact accttggtga tctcgccttt cacgtagtgg
     2641 acaaattctt ccaactgatc tgcgcgcgag gccaagcgat cttcttcttg tccaagataa
     2701 gcctgtctag cttcaagtat gacgggctga tactgggccg gcaggcgctc cattgcccag
     2761 tcggcagcga catccttcgg cgcgattttg ccggttactg cgctgtacca aatgcgggac
     2821 aacgtaagca ctacatttcg ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt
     2881 aaggtttcat ttagcgcctc aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc
     2941 gccgctggac ctaccaaggc aacgctatgt tctcttgctt ttgtcagcaa gatagccaga
     3001 tcaatgtcga tcgtggctgg ctcgaagata cctgcaagaa tgtcattgcg ctgccattct
     3061 ccaaattgca gttcgcgctt agctggataa cgccacggaa tgatgtcgtc gtgcacaaca
     3121 atggtgactt ctacagcgcg gagaatctcg ctctctccag gggaagccga agtttccaaa
     3181 aggtcgttga tcaaagctcg ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc
     3241 aaatcaatat cactgtgtgg cttcaggccg ccatccactg cggagccgta caaatgtacg
     3301 gccagcaacg tcggttcgag atggcgctcg atgacgccaa ctacctctga tagttgagtc
     3361 gatacttcgg cgatcaccgc ttccctcata atgtttaact ttgttttagg gcgactgccc
     3421 tgctgcgtaa catcgttgct gctccataac atcaaacatc gacccacggc gtaacgcgct
     3481 tgctgcttgg atgcccgagg catagactgt accccaaaaa aacagtcata acaagccatg
     3541 aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt
     3601 gagcgcatac gctacttgca ttacagctta cgaaccgaac aggcttatgt ccactgggtt
     3661 cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa ccttgggtag cagcgaagtc
     3721 gaggcatttc tgtcctggct ggtccagagg aggcatgatg atatattttt atcttgtgca
     3781 atgtaacatc agagattttg agacacgggc cagagctgcc aggaaacagc tatgaccatg
     3841 taatacgact cactataggg gatgctatga ccatgtaata cgactcacta taggggatat
     3901 cagctggatg gcaaataatg attttatttt gactgatagt gacctgttcg ttgcaacaaa
     3961 ttgatgagca attatttttt ataatgccaa ctttgtacaa gaaagctggg tcgaattccg
     4021 gccgggtacc gtcccattcg ccatgccgaa gcatgttgcc cagccggcgc cagcgaggag
     4081 gctgggacca tgccggccag agacgccgcg gtcagactga gctccgtctc atctacaaca
     4141 gtagaaatta gacgagctta ctcgtttcgt cctcacggac tcatcagtaa tttggatccc
     4201 tgcagaagta acaccaaaca acagggtgag catcgacaaa agaaacagta ccaagcaaat
     4261 aaatagcgta tgaaggcagg gctaaaaaaa tccacatata gctgctgcat atgccatcat
     4321 ccaagtatat caagatcaaa ataattataa aacatacttg tttattataa tagataggta
     4381 ctcaaggtta gagcatatga atagatgctg catatgccat catgtatatg catcagtaaa
     4441 acccacatca acatgtatac ctatcctaga tcgatatttc catccatctt aaactcgtaa
     4501 ctatgaagat gtatgacaca cacatacagt tccaaaatta ataaatacac caggtagttt
     4561 gaaacagtat tctactccga tctagaacga atgaacgacc gcccaaccac accacatcat
     4621 cacaaccaag cgaacaaaaa gcatctctgt atatgcatca gtaaaacccg catcaacatg
     4681 tatacctatc ctagatcgat atttccatcc atcatcttca attcgtaact atgaatatgt
     4741 atggcacaca catacagatc caaaattaat aaatccacca ggtagtttga aacagaatta
     4801 attctactcc gatctagaac gaccgcccaa ccagaccaca tcatcacaac caagacaaaa
     4861 aaaagcatga aaagatgacc cgacaaacaa gtgcacggca tatattgaaa taaaggaaaa
     4921 gggcaaacca aaccctatgc aacgaaacaa aaaaaatcat gaaatcgatc ccgtctgcgg
     4981 aacggctag
//
