LOCUS       sequence-226916-        8874 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A-to-G base editor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8874)
  AUTHORS   Huang TP, Zhao KT, Miller SM, Gaudelli NM, Oakes BL, Fellmann C, 
            Savage DF, Liu DR
  TITLE     Circularly permuted and PAM-modified Cas9 variants broaden the 
            targeting scope of base editors.
  JOURNAL   Nat Biotechnol. 2019 May 20. pii: 10.1038/s41587-019-0134-y. doi: 
            10.1038/s41587-019-0134-y.
  PUBMED    31110355
REFERENCE   2  (bases 1 to 8874)
  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: "Nat 
            Biotechnol. 2019 May 20. pii: 10.1038/s41587-019-0134-y. doi: 
            10.1038/s41587-019-0134-y."
COMMENT     Sequence Label: CP1028-ABEmax
FEATURES             Location/Qualifiers
     source          1..8874
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        132..335
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     286..306
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     377..396
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        377..395
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             445..465
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2737..4455
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
     CDS             5860..5880
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5889..5906
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5929..5946)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5935..6159
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6230..6246)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6230..6246)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6243..6265)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6254..6270
                     /label=lac operator
                     /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(join(6278..6284,6285..6302,6303..6308))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6323..6344
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6461..6478)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6632..7220)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6712..6731)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7391..8182,8183..8251))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8014..8033
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8252..8356)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8435..8454)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(8626..8874,1..131)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
       61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat gaaacggaca
      421 gccgacggaa gcgagttcga gtcaccaaag aagaagcgga aagtctctga agtcgagttt
      481 agccacgagt attggatgag gcacgcactg accctggcaa agcgagcatg ggatgaaaga
      541 gaagtccccg tgggcgccgt gctggtgcac aacaatagag tgatcggaga gggatggaac
      601 aggccaatcg gccgccacga ccctaccgca cacgcagaga tcatggcact gaggcaggga
      661 ggcctggtca tgcagaatta ccgcctgatc gatgccaccc tgtatgtgac actggagcca
      721 tgcgtgatgt gcgcaggagc aatgatccac agcaggatcg gaagagtggt gttcggagca
      781 cgggacgcca agaccggcgc agcaggctcc ctgatggatg tgctgcacca ccccggcatg
      841 aaccaccggg tggagatcac agagggaatc ctggcagacg agtgcgccgc cctgctgagc
      901 gatttcttta gaatgcggag acaggagatc aaggcccaga agaaggcaca gagctccacc
      961 gactctggag gatctagcgg aggatcctct ggaagcgaga caccaggcac aagcgagtcc
     1021 gccacaccag agagctccgg cggctcctcc ggaggatcct ctgaggtgga gttttcccac
     1081 gagtactgga tgagacatgc cctgaccctg gccaagaggg cacgcgatga gagggaggtg
     1141 cctgtgggag ccgtgctggt gctgaacaat agagtgatcg gcgagggctg gaacagagcc
     1201 atcggcctgc acgacccaac agcccatgcc gaaattatgg ccctgagaca gggcggcctg
     1261 gtcatgcaga actacagact gattgacgcc accctgtacg tgacattcga gccttgcgtg
     1321 atgtgcgccg gcgccatgat ccactctagg atcggccgcg tggtgtttgg cgtgaggaac
     1381 gcaaaaaccg gcgccgcagg ctccctgatg gacgtgctgc actaccccgg catgaatcac
     1441 cgcgtcgaaa ttaccgaggg aatcctggca gatgaatgtg ccgccctgct gtgctatttc
     1501 tttcggatgc ctagacaggt gttcaatgct cagaagaagg cccagagctc caccgactcc
     1561 ggaggatcta gcggaggctc ctctggctct gagacacctg gcacaagcga gagcgcaaca
     1621 cctgaaagca gcgggggcag cagcgggggg tcagaaatcg gcaaggctac cgccaagtac
     1681 ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag
     1741 atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag
     1801 ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa
     1861 aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc
     1921 gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc
     1981 cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa
     2041 ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag
     2101 aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc
     2161 aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct
     2221 gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg
     2281 tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag
     2341 ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc
     2401 tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag
     2461 caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc
     2521 aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac
     2581 accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac
     2641 gagacacgga tcgacctgtc tcagctggga ggcgacggcg gatcaggagg ctctggcggt
     2701 tcaggtggat caggcggtag cggaggttca ggtggtatgg acaagaagta cagcatcggc
     2761 ctggccatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc
     2821 agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc
     2881 ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc
     2941 agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac
     3001 gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa
     3061 gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac
     3121 cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag
     3181 gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc
     3241 ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg
     3301 gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc
     3361 aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag
     3421 ctgcccggcg agaagaagaa tggcctgttc ggaaacctga ttgccctgag cctgggcctg
     3481 acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag
     3541 gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac
     3601 ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg
     3661 aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac
     3721 caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa
     3781 gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc
     3841 caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa
     3901 ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc
     3961 agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat
     4021 ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc
     4081 ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag
     4141 agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc
     4201 cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg
     4261 cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa
     4321 tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc
     4381 gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac
     4441 ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac
     4501 gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac
     4561 aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac
     4621 agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg
     4681 aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac
     4741 ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc
     4801 gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc
     4861 cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc
     4921 ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg
     4981 aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag
     5041 accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc
     5101 aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac
     5161 gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg
     5221 gacatcaacc ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag
     5281 gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac
     5341 aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac
     5401 gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg
     5461 agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca
     5521 aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag
     5581 ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag
     5641 gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac
     5701 ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc
     5761 gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcagtct
     5821 ggcggctcaa aaagaaccgc cgacggcagc gaattcgagc ccaagaagaa gaggaaagtc
     5881 taaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc
     5941 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     6001 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     6061 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     6121 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     6181 gctggggctc gataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     6241 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6301 agtgtaaagc ctagggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6361 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6421 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     6481 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6541 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6601 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6661 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6721 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6781 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6841 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6901 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6961 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7021 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     7081 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7141 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7201 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7261 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7321 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7381 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7441 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7501 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7561 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7621 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7681 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7741 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7801 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7861 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7921 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7981 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     8041 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     8101 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     8161 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     8221 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8281 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8341 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     8401 atcgatctcc cgatccccta gggtcgactc tcagtacaat ctgctctgat gccgcatagt
     8461 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     8521 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
     8581 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga
     8641 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     8701 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     8761 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     8821 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatc
//
