LOCUS       dABE7.10                8727 bp ds-DNA     circular SYN 28-NOV-2018
DEFINITION  expresses ABE7.10 in mammalian cells.
ACCESSION
VERSION     .
KEYWORDS    pCMV-ABE7.10.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8727)
  AUTHORS   Gaudelli NM, Komor AC, Rees HA, Packer MS, Badran AH, Bryson DI and
            Liu DR
  TITLE     Programmable base editing of A*T to G*C in genomic DNA without DNA
            cleavage.
  JOURNAL   Nature. 2017 Nov 23;551(7681):464-471. doi: 10.1038/nature24644.
            Epub 2017 Oct 25.
  PUBMED    29160308
REFERENCE   2  (bases 1 to 8727)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8727
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        19..398
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        399..602
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     553..573
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        644..662
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     644..663
                     /note="T7 promoter, forward primer"
     CDS             1867..5967
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the
                      Streptococcus pyogenes Type II CRISPR/Cas system"
                     /note="generates RNA-guided single strand nicks in DNA due
                      to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEE
                     SFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHM
                     IKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKS
                     RRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDN
                     LLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLL
                     KALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKL
                     NREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYY
                     VGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLP
                     KHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKED
                     YFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTL
                     FEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFL
                     KSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTV
                     KVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEH
                     PVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVL
                     TRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAG
                     FIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFY
                     KVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGK
                     ATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSM
                     PQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVA
                     KVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFE
                     LENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHK
                     HYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAP
                     AAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD"
     misc_feature    4381..4382
                     /note="Geneious type: Editing History Replacement"
     misc_feature    complement(4381..4382)
     CDS             5980..6000
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian
                     virus 40) large T antigen"
                     /translation="PKKKRKV"
     CDS             6009..6026
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /translation="HHHHHH"
     primer_bind     complement(6049..6066)
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6055..6279
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6350..6366)
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6350..6366)
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6363..6385)
                     /note="In lacZ gene"
     protein_bind    6374..6390
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(6398..6428)
                     /note="promoter for the E. coli lac operon"
     protein_bind    6443..6464
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                      of cAMP."
     primer_bind     complement(6581..6598)
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6752..7340)
     primer_bind     complement(6832..6851)
                     /note="pBR322 origin, forward primer"
     CDS             complement(7511..8371)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     primer_bind     8134..8153
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8372..8476)
                     /gene="bla"
     primer_bind     complement(8555..8574)
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN      
        1 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
       61 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      121 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      181 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      241 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      301 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      361 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      421 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      481 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      541 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      601 ctggtttagt gaaccgtcag atccgctaga gatccgcggc cgctaatacg actcactata
      661 gggagagccg ccaccatgtc cgaagtcgag ttttcccatg agtactggat gagacacgca
      721 ttgactctcg caaagagggc ttgggatgaa cgcgaggtgc ccgtgggggc agtactcgtg
      781 cataacaatc gcgtaatcgg cgaaggttgg aataggccga tcggacgcca cgaccccact
      841 gcacatgcgg aaatcatggc ccttcgacag ggagggcttg tgatgcagaa ttatcgactt
      901 atcgatgcga cgctgtacgt cacgcttgaa ccttgcgtaa tgtgcgcggg agctatgatt
      961 cactcccgca ttggacgagt tgtattcggt gcccgcgacg ccaagacggg tgccgcaggt
     1021 tcactgatgg acgtgctgca tcacccaggc atgaaccacc gggtagaaat cacagaaggc
     1081 atattggcgg acgaatgtgc ggcgctgttg tccgactttt ttcgcatgcg gaggcaggag
     1141 atcaaggccc agaaaaaagc acaatcctct actgactctg gtggttcttc tggtggttct
     1201 agcggcagcg agactcccgg gacctcagag tccgccacac ccgaaagttc tggtggttct
     1261 tctggtggtt cttccgaagt cgagttttcc catgagtact ggatgagaca cgcattgact
     1321 ctcgcaaaga gggctcgaga tgaacgcgag gtgcccgtgg gggcagtact cgtgctcaac
     1381 aatcgcgtaa tcggcgaagg ttggaatagg gcaatcggac tccacgaccc cactgcacat
     1441 gcggaaatca tggcccttcg acagggaggg cttgtgatgc agaattatcg acttatcgat
     1501 gcgacgctgt acgtcacgtt tgaaccttgc gtaatgtgcg cgggagctat gattcactcc
     1561 cgcattggac gagttgtatt cggtgttcgc aacgccaaga cgggtgccgc aggttcactg
     1621 atggacgtgc tgcattaccc aggcatgaac caccgggtag aaatcacaga aggcatattg
     1681 gcggacgaat gtgcggcgct gttgtgttac ttttttcgca tgcccaggca ggtctttaac
     1741 gcccagaaaa aagcacaatc ctctactgac tctggtggtt cttctggtgg ttctagcggc
     1801 agcgagactc ccgggacctc agagtccgcc acacccgaaa gttctggtgg ttcttctggt
     1861 ggttctgata aaaagtattc tattggttta gccatcggca ctaattccgt tggatgggct
     1921 gtcataaccg atgaatacaa agtaccttca aagaaattta aggtgttggg gaacacagac
     1981 cgtcattcga ttaaaaagaa tcttatcggt gccctcctat tcgatagtgg cgaaacggca
     2041 gaggcgactc gcctgaaacg aaccgctcgg agaaggtata cacgtcgcaa gaaccgaata
     2101 tgttacttac aagaaatttt tagcaatgag atggccaaag ttgacgattc tttctttcac
     2161 cgtttggaag agtccttcct tgtcgaagag gacaagaaac atgaacggca ccccatcttt
     2221 ggaaacatag tagatgaggt ggcatatcat gaaaagtacc caacgattta tcacctcaga
     2281 aaaaagctag ttgactcaac tgataaagcg gacctgaggt taatctactt ggctcttgcc
     2341 catatgataa agttccgtgg gcactttctc attgagggtg atctaaatcc ggacaactcg
     2401 gatgtcgaca aactgttcat ccagttagta caaacctata atcagttgtt tgaagagaac
     2461 cctataaatg caagtggcgt ggatgcgaag gctattctta gcgcccgcct ctctaaatcc
     2521 cgacggctag aaaacctgat cgcacaatta cccggagaga agaaaaatgg gttgttcggt
     2581 aaccttatag cgctctcact aggcctgaca ccaaatttta agtcgaactt cgacttagct
     2641 gaagatgcca aattgcagct tagtaaggac acgtacgatg acgatctcga caatctactg
     2701 gcacaaattg gagatcagta tgcggactta tttttggctg ccaaaaacct tagcgatgca
     2761 atcctcctat ctgacatact gagagttaat actgagatta ccaaggcgcc gttatccgct
     2821 tcaatgatca aaaggtacga tgaacatcac caagacttga cacttctcaa ggccctagtc
     2881 cgtcagcaac tgcctgagaa atataaggaa atattctttg atcagtcgaa aaacgggtac
     2941 gcaggttata ttgacggcgg agcgagtcaa gaggaattct acaagtttat caaacccata
     3001 ttagagaaga tggatgggac ggaagagttg cttgtaaaac tcaatcgcga agatctactg
     3061 cgaaagcagc ggactttcga caacggtagc attccacatc aaatccactt aggcgaattg
     3121 catgctatac ttagaaggca ggaggatttt tatccgttcc tcaaagacaa tcgtgaaaag
     3181 attgagaaaa tcctaacctt tcgcatacct tactatgtgg gacccctggc ccgagggaac
     3241 tctcggttcg catggatgac aagaaagtcc gaagaaacga ttactccatg gaattttgag
     3301 gaagttgtcg ataaaggtgc gtcagctcaa tcgttcatcg agaggatgac caactttgac
     3361 aagaatttac cgaacgaaaa agtattgcct aagcacagtt tactttacga gtatttcaca
     3421 gtgtacaatg aactcacgaa agttaagtat gtcactgagg gcatgcgtaa acccgccttt
     3481 ctaagcggag aacagaagaa agcaatagta gatctgttat tcaagaccaa ccgcaaagtg
     3541 acagttaagc aattgaaaga ggactacttt aagaaaattg aatgcttcga ttctgtcgag
     3601 atctccgggg tagaagatcg atttaatgcg tcacttggta cgtatcatga cctcctaaag
     3661 ataattaaag ataaggactt cctggataac gaagagaatg aagatatctt agaagatata
     3721 gtgttgactc ttaccctctt tgaagatcgg gaaatgattg aggaaagact aaaaacatac
     3781 gctcacctgt tcgacgataa ggttatgaaa cagttaaaga ggcgtcgcta tacgggctgg
     3841 ggacgattgt cgcggaaact tatcaacggg ataagagaca agcaaagtgg taaaactatt
     3901 ctcgattttc taaagagcga cggcttcgcc aataggaact ttatgcagct gatccatgat
     3961 gactctttaa ccttcaaaga ggatatacaa aaggcacagg tttccggaca aggggactca
     4021 ttgcacgaac atattgcgaa tcttgctggt tcgccagcca tcaaaaaggg catactccag
     4081 acagtcaaag tagtggatga gctagttaag gtcatgggac gtcacaaacc ggaaaacatt
     4141 gtaatcgaga tggcacgcga aaatcaaacg actcagaagg ggcaaaaaaa cagtcgagag
     4201 cggatgaaga gaatagaaga gggtattaaa gaactgggca gccagatctt aaaggagcat
     4261 cctgtggaaa atacccaatt gcagaacgag aaactttacc tctattacct acaaaatgga
     4321 agggacatgt atgttgatca ggaactggac ataaaccgtt tatctgatta cgacgtcgat
     4381 gccattgtac cccaatcctt tttgaaggac gattcaatcg acaataaagt gcttacacgc
     4441 tcggataaga accgagggaa aagtgacaat gttccaagcg aggaagtcgt aaagaaaatg
     4501 aagaactatt ggcggcagct cctaaatgcg aaactgataa cgcaaagaaa gttcgataac
     4561 ttaactaaag ctgagagggg tggcttgtct gaacttgaca aggccggatt tattaaacgt
     4621 cagctcgtgg aaacccgcca aatcacaaag catgttgcac agatactaga ttcccgaatg
     4681 aatacgaaat acgacgagaa cgataagctg attcgggaag tcaaagtaat cactttaaag
     4741 tcaaaattgg tgtcggactt cagaaaggat tttcaattct ataaagttag ggagataaat
     4801 aactaccacc atgcgcacga cgcttatctt aatgccgtcg tagggaccgc actcattaag
     4861 aaatacccga agctagaaag tgagtttgtg tatggtgatt acaaagttta tgacgtccgt
     4921 aagatgatcg cgaaaagcga acaggagata ggcaaggcta cagccaaata cttcttttat
     4981 tctaacatta tgaatttctt taagacggaa atcactctgg caaacggaga gatacgcaaa
     5041 cgacctttaa ttgaaaccaa tggggagaca ggtgaaatcg tatgggataa gggccgggac
     5101 ttcgcgacgg tgagaaaagt tttgtccatg ccccaagtca acatagtaaa gaaaactgag
     5161 gtgcagaccg gagggttttc aaaggaatcg attcttccaa aaaggaatag tgataagctc
     5221 atcgctcgta aaaaggactg ggacccgaaa aagtacggtg gcttcgatag ccctacagtt
     5281 gcctattctg tcctagtagt ggcaaaagtt gagaagggaa aatccaagaa actgaagtca
     5341 gtcaaagaat tattggggat aacgattatg gagcgctcgt cttttgaaaa gaaccccatc
     5401 gacttccttg aggcgaaagg ttacaaggaa gtaaaaaagg atctcataat taaactacca
     5461 aagtatagtc tgtttgagtt agaaaatggc cgaaaacgga tgttggctag cgccggagag
     5521 cttcaaaagg ggaacgaact cgcactaccg tctaaatacg tgaatttcct gtatttagcg
     5581 tcccattacg agaagttgaa aggttcacct gaagataacg aacagaagca actttttgtt
     5641 gagcagcaca aacattatct cgacgaaatc atagagcaaa tttcggaatt cagtaagaga
     5701 gtcatcctag ctgatgccaa tctggacaaa gtattaagcg catacaacaa gcacagggat
     5761 aaacccatac gtgagcaggc ggaaaatatt atccatttgt ttactcttac caacctcggc
     5821 gctccagccg cattcaagta ttttgacaca acgatagatc gcaaacgata cacttctacc
     5881 aaggaggtgc tagacgcgac actgattcac caatccatca cgggattata tgaaactcgg
     5941 atagatttgt cacagcttgg gggtgactct ggtggttctc ccaagaagaa gaggaaagtc
     6001 taaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc
     6061 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     6121 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     6181 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     6241 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     6301 gctggggctc gataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     6361 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6421 agtgtaaagc ctagggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6481 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6541 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     6601 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6661 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6721 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6781 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6841 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6901 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6961 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     7021 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7081 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7141 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     7201 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7261 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7321 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7381 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7441 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7501 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7561 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7621 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7681 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7741 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7801 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7861 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7921 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7981 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     8041 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     8101 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8161 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8221 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8281 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8341 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8401 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8461 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     8521 atcgatctcc cgatccccta gggtcgactc tcagtacaat ctgctctgat gccgcatagt
     8581 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     8641 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
     8701 ggcgttttgc gctgcttcgc gatgtac
//
