LOCUS       pFH92                   7597 bp    DNA     circular UNA 10-APR-2019
DEFINITION  Level0 nScCas9-ABE7.10 base changer, wheat/human codon optimized.
ACCESSION   urn.local...1bx-abnc61t
VERSION     urn.local...1bx-abnc61t
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..2733
                     /created_by="hahnf"
                     /label="ecTadA (wt), monomer 1 of 2"
     ligation        <2233..2235
                     /label="Ligation"
     Linker          2734..2829
                     /created_by="hahnf"
                     /label="Linker"
     misc_feature    2758..2805
                     /created_by="hahnf"
                     /label="XTEN Linker"
     CDS             2830..3327
                     /created_by="hahnf"
                     /modified_by="hahnf"
                     /label="ecTadA (evolved), monomer 2 of 2"
     ligation        2836..2839
                     /label="Ligation"
     Linker          3328..3423
                     /created_by="hahnf"
                     /label="Linker"
     misc_feature    3352..3399
                     /created_by="hahnf"
                     /label="XTEN linker"
     CDS             3424..7578
                     /created_by="hahnf"
                     /modified_by="hahnf"
                     /label="ScCas9 nickase"
     ligation        3439..3442
                     /label="Ligation"
     mutation        3448..3450
                     /created_by="hahnf"
                     /label="D10A"
     ligation        5116..5119
                     /label="Ligation"
     sig_peptide     7558..7578
                     /created_by="hahnf"
                     /modified_by="hahnf"
                     /label="SV40 NLS"
     Stop_codon      7579..7581
                     /created_by="hahnf"
                     /label="STOP"
     ligation        7582..7585
                     /label="Ligation"
     misc_feature    complement(7587..7592)
                     /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 aaatgtccga agtcgagttt tcccatgagt actggatgag acacgcattg
     2281 actctcgcaa agagggcttg ggatgaacgc gaggtgcccg tgggggcagt actcgtgcat
     2341 aacaatcgcg taatcggcga aggttggaat aggccgatcg gacgccacga ccccactgca
     2401 catgcggaaa tcatggccct tcgacaggga gggcttgtga tgcagaatta tcgacttatc
     2461 gatgcgacgc tgtacgtcac gcttgaacct tgcgtaatgt gcgcgggagc tatgattcac
     2521 tcccgcattg gacgagttgt attcggtgcc cgcgacgcca agacgggtgc cgcaggttca
     2581 ctgatggacg tgctgcatca cccaggcatg aaccaccggg tagaaatcac agaaggcata
     2641 ttggcggacg aatgtgcggc gctgttgtcc gacttttttc gcatgcggag gcaggagatc
     2701 aaggcccaga aaaaagcaca atcctctact gactctggtg gttcttctgg tggttctagc
     2761 ggcagcgaga ctcccgggac ctcagagtcc gccacacccg aaagttctgg tggttcttct
     2821 ggtggttctt ccgaagtcga gttttcccat gagtactgga tgagacacgc attgactctc
     2881 gcaaagaggg ctcgagatga acgcgaggtg cccgtggggg cagtactcgt gctcaacaat
     2941 cgcgtaatcg gcgaaggttg gaatagggca atcggactcc acgaccccac tgcacatgcg
     3001 gaaatcatgg cccttcgaca gggagggctt gtgatgcaga attatcgact tatcgatgcg
     3061 acgctgtacg tcacgtttga accttgcgta atgtgcgcgg gagctatgat tcactcccgc
     3121 attggacgag ttgtattcgg tgttcgcaac gccaagacgg gtgccgcagg ttcactgatg
     3181 gacgtgctgc attacccagg catgaaccac cgggtagaaa tcacagaagg catattggcg
     3241 gacgaatgtg cggcgctgtt gtgttacttt tttcgcatgc ccaggcaggt ctttaacgcc
     3301 cagaaaaaag cacaatcctc tactgactct ggtggttctt ctggtggttc tagcggcagc
     3361 gagactcccg ggacctcaga gtccgccaca cccgaaagtt ctggtggttc ttctggtggt
     3421 tctgagaaaa agtattctat tgggctggcc attggcacca attccgttgg atgggccgtg
     3481 atcactgatg attataaggt cccctccaag aagttcaagg ttctgggtaa taccaacagg
     3541 aagagcataa aaaagaatct tatgggcgct ctgctctttg atagcggaga aacggcggaa
     3601 gctacacggc tcaagcggac agcgaggcgc cgctatacca ggcggaagaa ccggattaga
     3661 taccttcagg aaatcttcgc gaacgagatg gccaagctcg atgactcatt cttccagcgc
     3721 cttgaggaat cgttcctcgt tgaggaggac aaaaagaacg agcgtcaccc catctttggt
     3781 aatctcgcgg acgaggttgc ataccacagg aattatccga ccatctacca cctgaggaag
     3841 aagttggcgg acagcccgga gaaggcagac ctgcggctca tctacctcgc gctggcgcac
     3901 atcatcaaat tccgggggca ttttttgatc gaagggaaac tcaatgctga gaactcggac
     3961 gtggcaaagc tcttctatca gcttatccaa acctataatc agctgttcga ggaatctccg
     4021 ttggatgaga tagaggttga cgctaagggt atcctctccg ctcgtctttc gaagtccaaa
     4081 cgcctggaga aactgatcgc cgttttcccg aacgagaaga agaacgggct cttcggaaac
     4141 ataatcgcgc ttgccctggg actgacgccc aacttcaagt ccaacttcga cttgacggag
     4201 gatgccaaac ttcagctttc aaaggacaca tatgacgatg atttggacga gttgctcggc
     4261 caaattggcg accaatacgc tgacctgttc tctgccgcaa aaaacctgtc cgatgctatt
     4321 ctgctgagcg atattctccg gagcaactcc gaggtgacta aggcaccgtt gtccgccagc
     4381 atggtgaaac gctacgacga gcaccaccag gacctggctc tcctcaagac cttggttagg
     4441 caacagtttc ccgagaagta cgccgaaatc tttaaggatg acaccaagaa cggctatgcc
     4501 ggttacgttg gcatcggaat caagcatagg aagaggacca ctaaactcgc cacacaggag
     4561 gaattctata aatttatcaa gcctattctg gagaagatgg acggagctga ggagctgctc
     4621 gccaagctga acagggacga tctcctgagg aagcagcgga cctttgataa tggctcgatt
     4681 cctcatcaaa tccatctgaa ggagcttcat gccatactgc gcaggcaaga agagttttac
     4741 ccatttctga aggagaatcg ggagaagatc gaaaaaatcc tgacattccg cataccctac
     4801 tacgtgggcc cgcttgcccg gggaaactct cgttttgctt ggctcacgag gaagagcgag
     4861 gaggcgataa cgccatggaa cttcgaggag gtcgtggata aaggcgccag cgcgcaatcg
     4921 tttattgagc gtatgacaaa cttcgacgag cagctcccaa acaagaaggt gttgccaaag
     4981 cacagcctcc tgtacgagta cttcacggtt tacaatgagc tcaccaaagt caagtacgtt
     5041 actgagcgta tgcgtaagcc ggagttcctc tctggggagc agaagaaggc aatcgttgac
     5101 ctgctcttca aaacgaaccg taaggtgacc gtcaagcagc tgaaggaaga ttacttcaaa
     5161 aagatcgaat gttttgactc cgtggagatt ataggagttg aggataggtt taatgcctca
     5221 ctcggcacct accatgactt gctcaagatc atcaaagaca aggatttcct ggacaatgaa
     5281 gagaacgagg atatccttga ggacatcgtg ctcacactca ccctcttcga ggaccgggag
     5341 atgatcgagg agcgcctcaa gacatacgcc cacctctttg acgacaaggt gatgaaacaa
     5401 ctcaagcgca ggcactacac gggttgggga cgcctctccc gcaagatgat taatggtatc
     5461 agggacaaac agtccggtaa gacgattctc gatttcttga agtccgatgg tttctctaat
     5521 cgcaacttca tgcagctgat ccacgacgac tcactcacct tcaaggagga gattgaaaag
     5581 gctcaagtgt ccgggcaggg agatagcctc cacgagcaaa tcgcagatct cgccggatcg
     5641 cctgcaatta agaagggcat tcttcaaact gtcaaaatcg tcgacgagct ggtcaaggtt
     5701 atgggccaca agccagaaaa catcgttatc gagatggcta gggagaacca gaccactacc
     5761 aagggactcc aacagtcccg ggagcggaag aagaggattg aggaggggat caaggaactc
     5821 gagtctcaaa ttctcaagga gaatccggtg gagaacactc aactccagaa cgagaagctc
     5881 tacctgtact acctgcagaa cggcagggat atgtatgtcg atcaggagct ggacatcaac
     5941 aggctctccg actacgacgt cgaccatatc gtcccgcaat ctttcattaa ggacgactca
     6001 atcgataaca aggtcctcac gcgctccgtc gagaacagag gtaagtcaga caacgtgccg
     6061 tcggaggagg tggtgaagaa gatgaaaaac tactggcgtc aactcctcaa cgctaaattg
     6121 attacgcaga gaaaattcga taacttgacc aaagcggaac gcggcggtct gtcggaggcc
     6181 gacaaagccg gcttcattaa gcggcaactc gtggagacac gccagatcac aaagcacgtc
     6241 gccaggattc ttgactcgcg gatgaatact aaacgcgaca agaacgataa gccaatcaga
     6301 gaagtcaagg tgattacact gaagagcaaa ctggtcagcg acttccgcaa ggatttccag
     6361 ctttataagg tgagggacat aaacaattat caccacgccc acgacgcgta cctcaacgcg
     6421 gtggttggca ccgcactcat taagaaatac ccgaagctcg agtcagagtt cgtctacggc
     6481 gactataaag tgtacgacgt tcggaaaatg atagcaaagt ccgagcaaga gatcggcaag
     6541 gccacggcca agagattctt ctactccaat attatgaatt tctttaagac ggaggttaag
     6601 ttggccaacg gagagatccg gaagcggcct ctcattgaaa ctaatggcga gacaggcgag
     6661 gtggtctgga acaaggaaaa ggactttgct accgtccgca aggtcctcgc gatgccgcaa
     6721 gtgaacattg tcaaaaagac ggaagttcag acaggcgggt tctcgaagga gtcaatcttg
     6781 tcgaagcgtg agtccgccaa gctcatccca cgcaagaagg ggtgggatac gcggaagtat
     6841 ggaggtttcg ggtcccccac tgttgcatac tcaattctcg tggtcgccaa ggtggagaag
     6901 ggaaaagcca agaagctcaa aagcgtgaag gtcctcgtgg gtataacgat aatggaaaag
     6961 ggtagctacg agaaggaccc aatcggtttc ctcgaagcca aggggtacaa agatatcaaa
     7021 aaggagctta tattcaaact gccgaagtac tcattgtttg agctggagaa cggcaggcgg
     7081 cggatgttgg cctcggcgac agaactgcaa aaggcgaacg agctcgtctt gcctcagcat
     7141 ctcgttaggc ttctctacta cacccaaaat atctctgcga ctacagggtc taacaacctc
     7201 ggttacatcg agcagcatag ggaggaattc aaggagatct ttgagaagat catcgacttc
     7261 tccgagaagt acatccttaa gaacaaagtg aactccaact tgaagtcgag cttcgacgag
     7321 cagttcgctg tttccgattc tattctcctg tcgaactcat tcgtgtcact tctgaagtac
     7381 acgtcgttcg gcgcatctgg cggctttacc ttcctcgacc tcgacgtgaa gcaaggccgg
     7441 ctccgctacc aaaccgtgac cgaggttctc gacgcgacat tgatctacca gtccattacc
     7501 ggcctgtacg aaacgaggac ggacctctca cagctcggag gggactctag agcggaccca
     7561 aagaagaagc gcaaggtgta ggctttgaga ccacgaa
//
