LOCUS       pFH46                   6199 bp    DNA     circular UNA 18-NOV-2019
DEFINITION  Level0 FnCas12a, human codon optimized.
ACCESSION   urn.local...1w-aesjuvk
VERSION     urn.local...1w-aesjuvk
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     CDS             215..1225
                     /label="Sm/Sp no DraIII"
     rep_origin      1385..1999
                     /modified_by="hahnf"
                     /label="rep (pMB1)"
     rep_origin      1984
                     /label="ORI"
     misc_feature    2225..2230
                     /Recognition_pattern="GGTCTC(1/5)"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
     ligation        2232..2235
                     /label="Ligation"
     CDS             2233..6180
                     /created_by="hahnf"
                     /modified_by="hahnf"
                     /label="FnCas12a (human optimized)"
     sig_peptide     6133..6180
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /translation="KRPAATKKAGQAKKKK"
                     /modified_by="hahnf"
                     /label="Nucleoplasmin NLS"
     Stop_codon      6181..6183
                     /created_by="hahnf"
                     /label="Stop"
     ligation        6184..6187
                     /label="Ligation"
     misc_feature    complement(6189..6194)
                     /Recognition_pattern="GGTCTC(1/5)"
                     /label="BsaI"
                     /note="Geneious type: restriction site"
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 tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg
      361 tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc gccctaaaac
      421 aaagttaaac atcatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt
      481 agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc
      541 cgcagtggat ggcggcctga agccacacag cgatattgat ttgctggtta cggtgaccgt
      601 aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc
      661 ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat
      721 cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga
      781 cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac
      841 aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc
      901 ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc
      961 gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag
     1021 cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct
     1081 gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga
     1141 agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccattacg taaaaggcga
     1201 gatcaccaag gtagtcggca aataactgtc agaccaagtt tactcatata tactttagat
     1261 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     1321 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     1381 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1441 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1501 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     1561 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1621 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1681 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1741 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     1801 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     1861 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     1921 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     1981 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2041 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2101 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2161 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatcact
     2221 ctgtggtctc aaatgagcat ctaccaggag ttcgtcaaca agtattcact gagtaagaca
     2281 ctgcggttcg agctgatccc acagggcaag acactggaga acatcaaggc ccgaggcctg
     2341 attctggacg atgagaagcg ggcaaaagac tataagaaag ccaagcagat cattgataaa
     2401 taccaccagt tctttatcga ggaaattctg agctccgtgt gcatcagtga ggatctgctg
     2461 cagaattact cagacgtgta cttcaagctg aagaagagcg acgatgacaa cctgcagaag
     2521 gacttcaagt ccgccaagga caccatcaag aaacagatta gcgagtacat caaggactcc
     2581 gaaaagttta aaaatctgtt caaccagaat ctgatcgatg ctaagaaagg ccaggagtcc
     2641 gacctgatcc tgtggctgaa acagtctaag gacaatggga ttgaactgtt caaggctaac
     2701 tccgatatca ctgatattga cgaggcactg gaaatcatca agagcttcaa gggatggacc
     2761 acatacttta aaggcttcca cgagaaccgc aagaacgtgt actccagcaa cgacattcct
     2821 acctccatca tctaccgaat cgtcgatgac aatctgccaa agttcctgga gaacaaggcc
     2881 aaatatgaat ctctgaagga caaagctccc gaggcaatta attacgaaca gatcaagaaa
     2941 gatctggctg aggaactgac attcgatatc gactataaga ctagcgaggt gaaccagagg
     3001 gtcttttccc tggacgaggt gtttgaaatc gccaatttca acaattacct gaaccagtcc
     3061 ggcattacta aattcaatac catcattggc gggaagtttg tgaacgggga gaataccaag
     3121 cgcaagggaa ttaacgaata catcaatctg tatagccagc agatcaacga caaaactctg
     3181 aagaaataca agatgtctgt gctgttcaaa cagatcctga gtgataccga gtccaagtct
     3241 tttgtcattg ataaactgga agatgactca gacgtggtca ctaccatgca gagcttttat
     3301 gagcagatcg ccgctttcaa gacagtggag gaaaaatcta ttaaggaaac tctgagtctg
     3361 ctgttcgatg acctgaaagc ccagaagctg gacctgagta agatctactt caaaaacgat
     3421 aagagtctga cagacctgtc acagcaggtg tttgatgact attccgtgat tgggaccgcc
     3481 gtcctggagt acattacaca gcagatcgct ccaaagaacc tggataatcc ctctaagaaa
     3541 gagcaggaac tgatcgctaa gaaaaccgag aaggcaaaat atctgagtct ggaaacaatt
     3601 aagctggcac tggaggagtt caacaagcac agggatattg acaaacagtg ccgctttgag
     3661 gaaatcctgg ccaacttcgc agccatcccc atgatttttg atgagatcgc ccagaacaaa
     3721 gacaatctgg ctcagatcag tattaagtac cagaaccagg gcaagaaaga cctgctgcag
     3781 gcttcagcag aagatgacgt gaaagccatc aaggatctgc tggaccagac caacaatctg
     3841 ctgcacaagc tgaaaatctt ccatattagt cagtcagagg ataaggctaa tatcctggat
     3901 aaagacgaac acttctacct ggtgttcgag gaatgttact tcgagctggc aaacattgtc
     3961 cccctgtata acaagattag gaactacatc acacagaagc cttactctga cgagaagttt
     4021 aaactgaact tcgaaaatag taccctggcc aacgggtggg ataagaacaa ggagcctgac
     4081 aacacagcta tcctgttcat caaggatgac aagtactatc tgggagtgat gaataagaaa
     4141 aacaataaga tcttcgatga caaagccatt aaggagaaca aaggggaagg atacaagaaa
     4201 atcgtgtata agctgctgcc cggcgcaaat aagatgctgc ctaaggtgtt cttcagcgcc
     4261 aagagtatca aattctacaa cccatccgag gacatcctgc ggattagaaa tcactcaaca
     4321 catactaaga acgggagccc ccagaaggga tatgagaaat ttgagttcaa catcgaggat
     4381 tgcaggaagt ttattgactt ctacaagcag agcatctcca aacaccctga atggaaggat
     4441 tttggcttcc ggttttccga cacacagaga tataactcta tcgacgagtt ctaccgcgag
     4501 gtggaaaatc aggggtataa gctgactttt gagaacattt ctgaaagtta catcgacagc
     4561 gtggtcaatc agggaaagct gtacctgttc cagatctata acaaagattt ttcagcatac
     4621 agcaagggca gaccaaacct gcatacactg tactggaagg ccctgttcga tgagaggaat
     4681 ctgcaggacg tggtctataa actgaacgga gaggccgaac tgttttaccg gaagcagtct
     4741 attcctaaga aaatcactca cccagctaag gaggccatcg ctaacaagaa caaggacaat
     4801 cctaagaaag agagcgtgtt cgaatacgat ctgattaagg acaagcggtt caccgaagat
     4861 aagttctttt tccattgtcc aatcaccatt aacttcaagt caagcggcgc taacaagttc
     4921 aacgacgaga tcaatctgct gctgaaggaa aaagcaaacg atgtgcacat cctgagcatt
     4981 gaccgaggag agcggcatct ggcctactat accctggtgg atggcaaagg gaatatcatt
     5041 aagcaggata cattcaacat cattggcaat gaccggatga aaaccaacta ccacgataaa
     5101 ctggctgcaa tcgagaagga tagagactca gctaggaagg actggaagaa aatcaacaac
     5161 attaaggaga tgaaggaagg ctatctgagc caggtggtcc atgagattgc aaagctggtc
     5221 atcgaataca atgccattgt ggtgttcgag gatctgaact tcggctttaa gagggggcgc
     5281 tttaaggtgg aaaaacaggt ctatcagaag ctggagaaaa tgctgatcga aaagctgaat
     5341 tacctggtgt ttaaagataa cgagttcgac aagaccggag gcgtcctgag agcctaccag
     5401 ctgacagctc cctttgaaac tttcaagaaa atgggaaaac agacaggcat catctactat
     5461 gtgccagccg gattcacttc caagatctgc cccgtgaccg gctttgtcaa ccagctgtac
     5521 cctaaatatg agtcagtgag caagtcccag gaatttttca gcaagttcga taagatctgt
     5581 tataatctgg acaaggggta cttcgagttt tccttcgatt acaagaactt cggcgacaag
     5641 gccgctaagg ggaaatggac cattgcctcc ttcggatctc gcctgatcaa ctttcgaaat
     5701 tccgataaaa accacaattg ggacactagg gaggtgtacc caaccaagga gctggaaaag
     5761 ctgctgaaag actactctat cgagtatgga catggcgaat gcatcaaggc agccatctgt
     5821 ggcgagagtg ataagaaatt tttcgccaag ctgacctcag tgctgaatac aatcctgcag
     5881 atgcggaact caaagaccgg gacagaactg gactatctga ttagccccgt ggctgatgtc
     5941 aacggaaact tcttcgacag cagacaggca cccaaaaata tgcctcagga tgcagacgcc
     6001 aacggggcct accacatcgg gctgaaggga ctgatgctgc tgggccggat caagaacaat
     6061 caggagggga agaagctgaa cctggtcatt aagaacgagg aatacttcga gtttgtccag
     6121 aatagaaata acaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag
     6181 tgagctttga gaccacgaa
//
