LOCUS       sequence-252779-        7599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level0 nScCas9-ABE7.10, wheat/human codon optimized.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7599)
  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 7599)
  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: pFH92
FEATURES             Location/Qualifiers
     source          1..7599
                     /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             join(2235..2735,2736..2831,2832..3329,3330..3425)
                     /codon_start=1
                     /product="adenine base editor comprising wild-type E. coli 
                     TadA fused to the evolved TadA*(7.10) mutant (Gaudelli et 
                     al., 2017)"
                     /label=ABE(7.10)
                     /translation="MSEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVHNNRVIGEGW
                     NRPIGRHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTLEPCVMCAGAMIHSRIGRVVF
                     GARDAKTGAAGSLMDVLHHPGMNHRVEITEGILADECAALLSDFFRMRRQEIKAQKKAQ
                     SSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGSSEVEFSHEYWMRHALTLAKRARD
                     EREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVT
                     FEPCVMCAGAMIHSRIGRVVFGVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECA
                     ALLCYFFRMPRQVFNAQKKAQSSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGS"
     CDS             7560..7580
                     /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 gaagtcgagt tttcccatga gtactggatg agacacgcat
     2281 tgactctcgc aaagagggct tgggatgaac gcgaggtgcc cgtgggggca gtactcgtgc
     2341 ataacaatcg cgtaatcggc gaaggttgga ataggccgat cggacgccac gaccccactg
     2401 cacatgcgga aatcatggcc cttcgacagg gagggcttgt gatgcagaat tatcgactta
     2461 tcgatgcgac gctgtacgtc acgcttgaac cttgcgtaat gtgcgcggga gctatgattc
     2521 actcccgcat tggacgagtt gtattcggtg cccgcgacgc caagacgggt gccgcaggtt
     2581 cactgatgga cgtgctgcat cacccaggca tgaaccaccg ggtagaaatc acagaaggca
     2641 tattggcgga cgaatgtgcg gcgctgttgt ccgacttttt tcgcatgcgg aggcaggaga
     2701 tcaaggccca gaaaaaagca caatcctcta ctgactctgg tggttcttct ggtggttcta
     2761 gcggcagcga gactcccggg acctcagagt ccgccacacc cgaaagttct ggtggttctt
     2821 ctggtggttc ttccgaagtc gagttttccc atgagtactg gatgagacac gcattgactc
     2881 tcgcaaagag ggctcgagat gaacgcgagg tgcccgtggg ggcagtactc gtgctcaaca
     2941 atcgcgtaat cggcgaaggt tggaataggg caatcggact ccacgacccc actgcacatg
     3001 cggaaatcat ggcccttcga cagggagggc ttgtgatgca gaattatcga cttatcgatg
     3061 cgacgctgta cgtcacgttt gaaccttgcg taatgtgcgc gggagctatg attcactccc
     3121 gcattggacg agttgtattc ggtgttcgca acgccaagac gggtgccgca ggttcactga
     3181 tggacgtgct gcattaccca ggcatgaacc accgggtaga aatcacagaa ggcatattgg
     3241 cggacgaatg tgcggcgctg ttgtgttact tttttcgcat gcccaggcag gtctttaacg
     3301 cccagaaaaa agcacaatcc tctactgact ctggtggttc ttctggtggt tctagcggca
     3361 gcgagactcc cgggacctca gagtccgcca cacccgaaag ttctggtggt tcttctggtg
     3421 gttctgagaa aaagtattct attgggctgg ccattggcac caattccgtt ggatgggccg
     3481 tgatcactga tgattataag gtcccctcca agaagttcaa ggttctgggt aataccaaca
     3541 ggaagagcat aaaaaagaat cttatgggcg ctctgctctt tgatagcgga gaaacggcgg
     3601 aagctacacg gctcaagcgg acagcgaggc gccgctatac caggcggaag aaccggatta
     3661 gataccttca ggaaatcttc gcgaacgaga tggccaagct cgatgactca ttcttccagc
     3721 gccttgagga atcgttcctc gttgaggagg acaaaaagaa cgagcgtcac cccatctttg
     3781 gtaatctcgc ggacgaggtt gcataccaca ggaattatcc gaccatctac cacctgagga
     3841 agaagttggc ggacagcccg gagaaggcag acctgcggct catctacctc gcgctggcgc
     3901 acatcatcaa attccggggg cattttttga tcgaagggaa actcaatgct gagaactcgg
     3961 acgtggcaaa gctcttctat cagcttatcc aaacctataa tcagctgttc gaggaatctc
     4021 cgttggatga gatagaggtt gacgctaagg gtatcctctc cgctcgtctt tcgaagtcca
     4081 aacgcctgga gaaactgatc gccgttttcc cgaacgagaa gaagaacggg ctcttcggaa
     4141 acataatcgc gcttgccctg ggactgacgc ccaacttcaa gtccaacttc gacttgacgg
     4201 aggatgccaa acttcagctt tcaaaggaca catatgacga tgatttggac gagttgctcg
     4261 gccaaattgg cgaccaatac gctgacctgt tctctgccgc aaaaaacctg tccgatgcta
     4321 ttctgctgag cgatattctc cggagcaact ccgaggtgac taaggcaccg ttgtccgcca
     4381 gcatggtgaa acgctacgac gagcaccacc aggacctggc tctcctcaag accttggtta
     4441 ggcaacagtt tcccgagaag tacgccgaaa tctttaagga tgacaccaag aacggctatg
     4501 ccggttacgt tggcatcgga atcaagcata ggaagaggac cactaaactc gccacacagg
     4561 aggaattcta taaatttatc aagcctattc tggagaagat ggacggagct gaggagctgc
     4621 tcgccaagct gaacagggac gatctcctga ggaagcagcg gacctttgat aatggctcga
     4681 ttcctcatca aatccatctg aaggagcttc atgccatact gcgcaggcaa gaagagtttt
     4741 acccatttct gaaggagaat cgggagaaga tcgaaaaaat cctgacattc cgcataccct
     4801 actacgtggg cccgcttgcc cggggaaact ctcgttttgc ttggctcacg aggaagagcg
     4861 aggaggcgat aacgccatgg aacttcgagg aggtcgtgga taaaggcgcc agcgcgcaat
     4921 cgtttattga gcgtatgaca aacttcgacg agcagctccc aaacaagaag gtgttgccaa
     4981 agcacagcct cctgtacgag tacttcacgg tttacaatga gctcaccaaa gtcaagtacg
     5041 ttactgagcg tatgcgtaag ccggagttcc tctctgggga gcagaagaag gcaatcgttg
     5101 acctgctctt caaaacgaac cgtaaggtga ccgtcaagca gctgaaggaa gattacttca
     5161 aaaagatcga atgttttgac tccgtggaga ttataggagt tgaggatagg tttaatgcct
     5221 cactcggcac ctaccatgac ttgctcaaga tcatcaaaga caaggatttc ctggacaatg
     5281 aagagaacga ggatatcctt gaggacatcg tgctcacact caccctcttc gaggaccggg
     5341 agatgatcga ggagcgcctc aagacatacg cccacctctt tgacgacaag gtgatgaaac
     5401 aactcaagcg caggcactac acgggttggg gacgcctctc ccgcaagatg attaatggta
     5461 tcagggacaa acagtccggt aagacgattc tcgatttctt gaagtccgat ggtttctcta
     5521 atcgcaactt catgcagctg atccacgacg actcactcac cttcaaggag gagattgaaa
     5581 aggctcaagt gtccgggcag ggagatagcc tccacgagca aatcgcagat ctcgccggat
     5641 cgcctgcaat taagaagggc attcttcaaa ctgtcaaaat cgtcgacgag ctggtcaagg
     5701 ttatgggcca caagccagaa aacatcgtta tcgagatggc tagggagaac cagaccacta
     5761 ccaagggact ccaacagtcc cgggagcgga agaagaggat tgaggagggg atcaaggaac
     5821 tcgagtctca aattctcaag gagaatccgg tggagaacac tcaactccag aacgagaagc
     5881 tctacctgta ctacctgcag aacggcaggg atatgtatgt cgatcaggag ctggacatca
     5941 acaggctctc cgactacgac gtcgaccata tcgtcccgca atctttcatt aaggacgact
     6001 caatcgataa caaggtcctc acgcgctccg tcgagaacag aggtaagtca gacaacgtgc
     6061 cgtcggagga ggtggtgaag aagatgaaaa actactggcg tcaactcctc aacgctaaat
     6121 tgattacgca gagaaaattc gataacttga ccaaagcgga acgcggcggt ctgtcggagg
     6181 ccgacaaagc cggcttcatt aagcggcaac tcgtggagac acgccagatc acaaagcacg
     6241 tcgccaggat tcttgactcg cggatgaata ctaaacgcga caagaacgat aagccaatca
     6301 gagaagtcaa ggtgattaca ctgaagagca aactggtcag cgacttccgc aaggatttcc
     6361 agctttataa ggtgagggac ataaacaatt atcaccacgc ccacgacgcg tacctcaacg
     6421 cggtggttgg caccgcactc attaagaaat acccgaagct cgagtcagag ttcgtctacg
     6481 gcgactataa agtgtacgac gttcggaaaa tgatagcaaa gtccgagcaa gagatcggca
     6541 aggccacggc caagagattc ttctactcca atattatgaa tttctttaag acggaggtta
     6601 agttggccaa cggagagatc cggaagcggc ctctcattga aactaatggc gagacaggcg
     6661 aggtggtctg gaacaaggaa aaggactttg ctaccgtccg caaggtcctc gcgatgccgc
     6721 aagtgaacat tgtcaaaaag acggaagttc agacaggcgg gttctcgaag gagtcaatct
     6781 tgtcgaagcg tgagtccgcc aagctcatcc cacgcaagaa ggggtgggat acgcggaagt
     6841 atggaggttt cgggtccccc actgttgcat actcaattct cgtggtcgcc aaggtggaga
     6901 agggaaaagc caagaagctc aaaagcgtga aggtcctcgt gggtataacg ataatggaaa
     6961 agggtagcta cgagaaggac ccaatcggtt tcctcgaagc caaggggtac aaagatatca
     7021 aaaaggagct tatattcaaa ctgccgaagt actcattgtt tgagctggag aacggcaggc
     7081 ggcggatgtt ggcctcggcg acagaactgc aaaaggcgaa cgagctcgtc ttgcctcagc
     7141 atctcgttag gcttctctac tacacccaaa atatctctgc gactacaggg tctaacaacc
     7201 tcggttacat cgagcagcat agggaggaat tcaaggagat ctttgagaag atcatcgact
     7261 tctccgagaa gtacatcctt aagaacaaag tgaactccaa cttgaagtcg agcttcgacg
     7321 agcagttcgc tgtttccgat tctattctcc tgtcgaactc attcgtgtca cttctgaagt
     7381 acacgtcgtt cggcgcatct ggcggcttta ccttcctcga cctcgacgtg aagcaaggcc
     7441 ggctccgcta ccaaaccgtg accgaggttc tcgacgcgac attgatctac cagtccatta
     7501 ccggcctgta cgaaacgagg acggacctct cacagctcgg aggggactct agagcggacc
     7561 caaagaagaa gcgcaaggtg taggctttga gaccacgaa
//
