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 //