LOCUS       sequence-252768-        6201 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level0 FnCas12a, human codon optimized.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6201)
  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 6201)
  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: pFH46
FEATURES             Location/Qualifiers
     source          1..6201
                     /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             6135..6182
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
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 tcaaatgagc atctaccagg agttcgtcaa caagtattca ctgagtaaga
     2281 cactgcggtt cgagctgatc ccacagggca agacactgga gaacatcaag gcccgaggcc
     2341 tgattctgga cgatgagaag cgggcaaaag actataagaa agccaagcag atcattgata
     2401 aataccacca gttctttatc gaggaaattc tgagctccgt gtgcatcagt gaggatctgc
     2461 tgcagaatta ctcagacgtg tacttcaagc tgaagaagag cgacgatgac aacctgcaga
     2521 aggacttcaa gtccgccaag gacaccatca agaaacagat tagcgagtac atcaaggact
     2581 ccgaaaagtt taaaaatctg ttcaaccaga atctgatcga tgctaagaaa ggccaggagt
     2641 ccgacctgat cctgtggctg aaacagtcta aggacaatgg gattgaactg ttcaaggcta
     2701 actccgatat cactgatatt gacgaggcac tggaaatcat caagagcttc aagggatgga
     2761 ccacatactt taaaggcttc cacgagaacc gcaagaacgt gtactccagc aacgacattc
     2821 ctacctccat catctaccga atcgtcgatg acaatctgcc aaagttcctg gagaacaagg
     2881 ccaaatatga atctctgaag gacaaagctc ccgaggcaat taattacgaa cagatcaaga
     2941 aagatctggc tgaggaactg acattcgata tcgactataa gactagcgag gtgaaccaga
     3001 gggtcttttc cctggacgag gtgtttgaaa tcgccaattt caacaattac ctgaaccagt
     3061 ccggcattac taaattcaat accatcattg gcgggaagtt tgtgaacggg gagaatacca
     3121 agcgcaaggg aattaacgaa tacatcaatc tgtatagcca gcagatcaac gacaaaactc
     3181 tgaagaaata caagatgtct gtgctgttca aacagatcct gagtgatacc gagtccaagt
     3241 cttttgtcat tgataaactg gaagatgact cagacgtggt cactaccatg cagagctttt
     3301 atgagcagat cgccgctttc aagacagtgg aggaaaaatc tattaaggaa actctgagtc
     3361 tgctgttcga tgacctgaaa gcccagaagc tggacctgag taagatctac ttcaaaaacg
     3421 ataagagtct gacagacctg tcacagcagg tgtttgatga ctattccgtg attgggaccg
     3481 ccgtcctgga gtacattaca cagcagatcg ctccaaagaa cctggataat ccctctaaga
     3541 aagagcagga actgatcgct aagaaaaccg agaaggcaaa atatctgagt ctggaaacaa
     3601 ttaagctggc actggaggag ttcaacaagc acagggatat tgacaaacag tgccgctttg
     3661 aggaaatcct ggccaacttc gcagccatcc ccatgatttt tgatgagatc gcccagaaca
     3721 aagacaatct ggctcagatc agtattaagt accagaacca gggcaagaaa gacctgctgc
     3781 aggcttcagc agaagatgac gtgaaagcca tcaaggatct gctggaccag accaacaatc
     3841 tgctgcacaa gctgaaaatc ttccatatta gtcagtcaga ggataaggct aatatcctgg
     3901 ataaagacga acacttctac ctggtgttcg aggaatgtta cttcgagctg gcaaacattg
     3961 tccccctgta taacaagatt aggaactaca tcacacagaa gccttactct gacgagaagt
     4021 ttaaactgaa cttcgaaaat agtaccctgg ccaacgggtg ggataagaac aaggagcctg
     4081 acaacacagc tatcctgttc atcaaggatg acaagtacta tctgggagtg atgaataaga
     4141 aaaacaataa gatcttcgat gacaaagcca ttaaggagaa caaaggggaa ggatacaaga
     4201 aaatcgtgta taagctgctg cccggcgcaa ataagatgct gcctaaggtg ttcttcagcg
     4261 ccaagagtat caaattctac aacccatccg aggacatcct gcggattaga aatcactcaa
     4321 cacatactaa gaacgggagc ccccagaagg gatatgagaa atttgagttc aacatcgagg
     4381 attgcaggaa gtttattgac ttctacaagc agagcatctc caaacaccct gaatggaagg
     4441 attttggctt ccggttttcc gacacacaga gatataactc tatcgacgag ttctaccgcg
     4501 aggtggaaaa tcaggggtat aagctgactt ttgagaacat ttctgaaagt tacatcgaca
     4561 gcgtggtcaa tcagggaaag ctgtacctgt tccagatcta taacaaagat ttttcagcat
     4621 acagcaaggg cagaccaaac ctgcatacac tgtactggaa ggccctgttc gatgagagga
     4681 atctgcagga cgtggtctat aaactgaacg gagaggccga actgttttac cggaagcagt
     4741 ctattcctaa gaaaatcact cacccagcta aggaggccat cgctaacaag aacaaggaca
     4801 atcctaagaa agagagcgtg ttcgaatacg atctgattaa ggacaagcgg ttcaccgaag
     4861 ataagttctt tttccattgt ccaatcacca ttaacttcaa gtcaagcggc gctaacaagt
     4921 tcaacgacga gatcaatctg ctgctgaagg aaaaagcaaa cgatgtgcac atcctgagca
     4981 ttgaccgagg agagcggcat ctggcctact ataccctggt ggatggcaaa gggaatatca
     5041 ttaagcagga tacattcaac atcattggca atgaccggat gaaaaccaac taccacgata
     5101 aactggctgc aatcgagaag gatagagact cagctaggaa ggactggaag aaaatcaaca
     5161 acattaagga gatgaaggaa ggctatctga gccaggtggt ccatgagatt gcaaagctgg
     5221 tcatcgaata caatgccatt gtggtgttcg aggatctgaa cttcggcttt aagagggggc
     5281 gctttaaggt ggaaaaacag gtctatcaga agctggagaa aatgctgatc gaaaagctga
     5341 attacctggt gtttaaagat aacgagttcg acaagaccgg aggcgtcctg agagcctacc
     5401 agctgacagc tccctttgaa actttcaaga aaatgggaaa acagacaggc atcatctact
     5461 atgtgccagc cggattcact tccaagatct gccccgtgac cggctttgtc aaccagctgt
     5521 accctaaata tgagtcagtg agcaagtccc aggaattttt cagcaagttc gataagatct
     5581 gttataatct ggacaagggg tacttcgagt tttccttcga ttacaagaac ttcggcgaca
     5641 aggccgctaa ggggaaatgg accattgcct ccttcggatc tcgcctgatc aactttcgaa
     5701 attccgataa aaaccacaat tgggacacta gggaggtgta cccaaccaag gagctggaaa
     5761 agctgctgaa agactactct atcgagtatg gacatggcga atgcatcaag gcagccatct
     5821 gtggcgagag tgataagaaa tttttcgcca agctgacctc agtgctgaat acaatcctgc
     5881 agatgcggaa ctcaaagacc gggacagaac tggactatct gattagcccc gtggctgatg
     5941 tcaacggaaa cttcttcgac agcagacagg cacccaaaaa tatgcctcag gatgcagacg
     6001 ccaacggggc ctaccacatc gggctgaagg gactgatgct gctgggccgg atcaagaaca
     6061 atcaggaggg gaagaagctg aacctggtca ttaagaacga ggaatacttc gagtttgtcc
     6121 agaatagaaa taacaaaagg ccggcggcca cgaaaaaggc cggccaggca aaaaagaaaa
     6181 agtgagcttt gagaccacga a
//
