LOCUS       sequence-251817-        6189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level0 FnCas12a, rice codon optimized.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6189)
  AUTHORS   Hahn F, Korolev A, Sanjurjo Loures L, Nekrasov V
  TITLE     A modular cloning toolkit for genome editing in plants.
  JOURNAL   BMC Plant Biol. 2020 Apr 23;20(1):179. doi: 
            10.1186/s12870-020-02388-2.
  PUBMED    32326901
REFERENCE   2  (bases 1 to 6189)
  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"; journalName: "BMC Plant 
            Biol."; date: "2020-04-23"; volume: "20(1):179. doi"; pages: " 
            10.1186/s12870-020-02388-2"
COMMENT     Sequence Label: pFH16
FEATURES             Location/Qualifiers
     source          1..6189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(24..42)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        110..214
                     /gene="bla"
                     /label=AmpR promoter
     CDS             436..1227
                     /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      1398..1986
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1887..1906
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2140..2157
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             6150..6170
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 gtggctcttc agtggacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
       61 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
      121 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
      181 ataaatgctt caataatatt gaaaaaggaa gagtatgcgc tcacgcaact ggtccagaac
      241 cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg
      301 tttttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg
      361 ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa
      421 acaaagttaa acatcatgag ggaagcggtg atcgccgaag tatcgactca actatcagag
      481 gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc
      541 tccgcagtgg atggcggcct gaagccacac agcgatattg atttgctggt tacggtgacc
      601 gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct
      661 tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac
      721 atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat
      781 gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg
      841 acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat
      901 ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg
      961 ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac
     1021 agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc
     1081 ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa
     1141 gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccatta cgtaaaaggc
     1201 gagatcacca aggtagtcgg caaataactg tcagaccaag tttactcata tatactttag
     1261 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     1321 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1381 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1441 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1501 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     1561 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1621 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1681 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     1741 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     1801 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     1861 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     1921 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     1981 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2041 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2101 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2161 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatca
     2221 ctctgtggtc tcaaatgtcc atctaccaag agttcgtgaa caagtactcc ctgtccaaga
     2281 ccctccgctt cgagcttatc ccacagggca agacgctcga gaacatcaag gccaggggcc
     2341 tcatcctcga cgacgagaag agagccaagg actacaagaa ggccaagcag atcatcgaca
     2401 agtaccacca gttctttatc gaggagatcc tctcctccgt ctgcatctcc gaggacctgc
     2461 tccagaacta ctccgacgtg tacttcaagc tcaagaagtc cgacgacgac aacctccaga
     2521 aggacttcaa gtccgccaag gacaccatca agaagcagat ctccgagtac attaaggact
     2581 ccgagaagtt caagaacctc ttcaaccaga acctcatcga cgccaagaag gggcaagagt
     2641 ccgacctcat cctgtggctc aagcagtcca aggacaacgg catcgagctg ttcaaggcca
     2701 actccgacat caccgacatc gacgaggccc tcgagatcat taagtccttc aagggctgga
     2761 ccacctactt caaggggttc cacgagaacc gcaagaacgt gtactcctcc aacgacatcc
     2821 cgacctccat catctacagg atcgtggacg ataacctgcc gaagttcctt gagaacaagg
     2881 ccaagtacga gtccctcaag gacaaggccc cagaggccat caactacgag cagatcaaga
     2941 aggacctcgc cgaggagctg accttcgaca tcgattacaa gacctccgag gtgaaccagc
     3001 gcgtgttctc actcgacgag gtgttcgaga tcgccaactt caacaactac ctcaaccagt
     3061 ccggcatcac caagttcaac accatcatcg gcggcaagtt cgtcaacggc gagaacacaa
     3121 agcgcaaggg catcaacgag tacatcaacc tctactccca gcagattaac gacaagaccc
     3181 tgaagaagta caagatgtcc gtcctcttca agcaaatcct ctccgacacc gagtccaagt
     3241 ccttcgtgat cgacaagctc gaggacgaca gcgacgtggt gaccaccatg cagtccttct
     3301 acgagcaaat cgccgccttc aagaccgtcg aggagaagtc catcaaggag actctcagcc
     3361 tcctgttcga cgacctcaag gcccagaagc tcgacctctc caagatctat ttcaagaacg
     3421 ataagtccct caccgacctc agccagcagg ttttcgacga ctactccgtg atcggcacag
     3481 ccgtgctcga gtatatcacg cagcagatcg ccccgaagaa cctcgacaac ccgagcaaga
     3541 aggagcaaga gctgatcgcc aaaaagaccg agaaggcgaa gtacctcagc ctcgagacaa
     3601 tcaagctcgc cctcgaggag ttcaacaagc accgcgacat tgacaagcag tgccgctttg
     3661 aggagattct cgcgaacttc gccgcgatcc cgatgatctt cgatgagatc gcccagaaca
     3721 aggataacct cgcccagatc tccatcaagt accagaacca ggggaagaag gatcttctcc
     3781 aggcctccgc cgaggacgat gtgaaggcta ttaaggacct cctcgaccag accaacaacc
     3841 tcctccacaa gcttaagatc ttccacatct cccagtccga ggataaggcc aacatcctgg
     3901 acaaggacga gcacttctac ctcgtgttcg aggagtgcta cttcgagctg gcgaacatcg
     3961 tgccgctcta caacaagatc cgcaactaca tcacccagaa gccgtactcg gacgagaagt
     4021 ttaagctcaa cttcgagaac tccaccctcg ccaacggctg ggacaagaac aaggagccag
     4081 acaacaccgc catcctcttc atcaaggatg acaagtacta cctcggcgtg atgaacaaga
     4141 agaacaacaa gattttcgac gataaggcga tcaaggagaa caagggcgag ggctacaaga
     4201 agatcgtcta caagctcctc ccaggcgcca acaagatgct cccgaaggtg ttcttcagcg
     4261 ccaagagcat caagttctac aacccgtccg aggacatcct ccgcatccgc aaccattcca
     4321 cccacaccaa gaatggctcc ccgcagaagg gctacgagaa gttcgagttc aacatcgagg
     4381 actgccgcaa gttcatcgac ttctacaagc agagcatctc caagcaccca gagtggaagg
     4441 atttcggctt ccgcttcagc gacacccagc gctacaacag catcgatgag ttctaccgcg
     4501 aggtcgagaa tcagggctat aagctgacgt tcgagaatat ctccgagagc tacatcgact
     4561 ccgtggtcaa ccagggcaag ctctacctgt tccagatcta taacaaggat ttctccgcct
     4621 acagcaaggg caggccgaac ctccataccc tctactggaa ggccctcttc gacgagagga
     4681 acctccagga cgtggtgtac aagctcaatg gcgaggcgga gctgttctac cggaagcagt
     4741 ctatcccgaa gaagatcacc cacccggcca aggaggccat tgccaacaag aacaaggaca
     4801 acccgaagaa ggagtccgtt ttcgagtacg acctgatcaa ggataagcgc tttaccgagg
     4861 acaagttctt cttccactgc ccgatcacca tcaacttcaa gagcagcggc gcgaacaagt
     4921 tcaatgacga gatcaacctc ctgctgaagg agaaggctaa cgacgtccac atcctctcca
     4981 tcgacagggg cgagagacac ctcgcctact acacactcgt ggacggcaag ggcaacatca
     5041 tcaagcagga caccttcaat atcatcggca acgaccgcat gaaaaccaac taccacgata
     5101 agctggccgc catcgagaag gacagggatt ccgctaggaa ggactggaag aagattaaca
     5161 atattaagga gatgaaggag ggctacctct cccaggtggt gcacgagatt gccaagctcg
     5221 tcatcgagta caacgccatc gtggttttcg aggaccttaa cttcggcttc aagcgcggca
     5281 ggttcaaggt tgagaagcag gtctaccaaa agctcgagaa gatgctcatt gagaagctca
     5341 attacctggt gtttaaggat aacgagttcg acaagacagg cggcgtgctc agagcctacc
     5401 aactcacagc tccgtttgag acattcaaga agatgggcaa gcagaccggc attatctact
     5461 acgtgccagc cggcttcacc agcaagatct gcccagtgac cggcttcgtc aaccagctct
     5521 acccgaagta cgagagcgtg tccaagtcgc aagagttctt tagcaagttc gataagatct
     5581 gctacaacct cgataagggc tactttgagt tcagcttcga ctataagaat ttcggcgaca
     5641 aggccgcgaa gggcaagtgg acaattgctt ccttcggctc ccgcctgatc aacttccgca
     5701 acagcgacaa gaaccacaac tgggacaccc gcgaggtgta ccccacaaag gagttggaga
     5761 agctcctgaa ggactactcc attgagtacg gccacggcga gtgcatcaag gctgccattt
     5821 gcggcgagtc cgataagaag ttcttcgcca agctcacctc cgtgctcaac acgatcctcc
     5881 agatgcgcaa ctccaagacc ggcaccgagc tggactacct catttcccca gtggccgacg
     5941 tgaacggcaa cttcttcgat tctaggcagg ccccaaagaa catgccgcag gatgccgatg
     6001 ccaacggcgc ctatcatatt ggcctcaagg gcctcatgct cctgggccgc atcaagaaca
     6061 atcaagaggg caagaagctc aacctcgtca tcaagaacga ggagtacttc gagttcgtgc
     6121 agaaccggaa caacgactcc agggccgacc caaagaagaa gaggaaggtc tgagctttga
     6181 gaccacgaa
//
