LOCUS       sequence-231807-        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"
     enhancer        182..561
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        562..765
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     716..736
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     807..826
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        807..825
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             875..895
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3167..4885
                     /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             6290..6310
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6319..6336
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6359..6376)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6365..6589
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6660..6676)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6660..6676)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6673..6695)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6684..6700
                     /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(6708..6714,6715..6732,6733..6738))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6753..6774
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6891..6908)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7062..7650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7142..7161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7821..8612,8613..8681))
                     /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     8444..8463
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8682..8786)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(8865..8874,1..10))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
       61 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
      121 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
      181 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
      241 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      301 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      361 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      421 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      481 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      541 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      601 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      661 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      721 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
      781 cagatccgct agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat
      841 gaaacggaca gccgacggaa gcgagttcga gtcaccaaag aagaagcgga aagtctctga
      901 agtcgagttt agccacgagt attggatgag gcacgcactg accctggcaa agcgagcatg
      961 ggatgaaaga gaagtccccg tgggcgccgt gctggtgcac aacaatagag tgatcggaga
     1021 gggatggaac aggccaatcg gccgccacga ccctaccgca cacgcagaga tcatggcact
     1081 gaggcaggga ggcctggtca tgcagaatta ccgcctgatc gatgccaccc tgtatgtgac
     1141 actggagcca tgcgtgatgt gcgcaggagc aatgatccac agcaggatcg gaagagtggt
     1201 gttcggagca cgggacgcca agaccggcgc agcaggctcc ctgatggatg tgctgcacca
     1261 ccccggcatg aaccaccggg tggagatcac agagggaatc ctggcagacg agtgcgccgc
     1321 cctgctgagc gatttcttta gaatgcggag acaggagatc aaggcccaga agaaggcaca
     1381 gagctccacc gactctggag gatctagcgg aggatcctct ggaagcgaga caccaggcac
     1441 aagcgagtcc gccacaccag agagctccgg cggctcctcc ggaggatcct ctgaggtgga
     1501 gttttcccac gagtactgga tgagacatgc cctgaccctg gccaagaggg cacgcgatga
     1561 gagggaggtg cctgtgggag ccgtgctggt gctgaacaat agagtgatcg gcgagggctg
     1621 gaacagagcc atcggcctgc acgacccaac agcccatgcc gaaattatgg ccctgagaca
     1681 gggcggcctg gtcatgcaga actacagact gattgacgcc accctgtacg tgacattcga
     1741 gccttgcgtg atgtgcgccg gcgccatgat ccactctagg atcggccgcg tggtgtttgg
     1801 cgtgaggaac gcaaaaaccg gcgccgcagg ctccctgatg gacgtgctgc actaccccgg
     1861 catgaatcac cgcgtcgaaa ttaccgaggg aatcctggca gatgaatgtg ccgccctgct
     1921 gtgctatttc tttcggatgc ctagacaggt gttcaatgct cagaagaagg cccagagctc
     1981 caccgactcc ggaggatcta gcggaggctc ctctggctct gagacacctg gcacaagcga
     2041 gagcgcaaca cctgaaagca gcgggggcag cagcgggggg tcagaaatcg gcaaggctac
     2101 cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc
     2161 caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt
     2221 gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa
     2281 tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa
     2341 gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg
     2401 cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa
     2461 gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag
     2521 cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga
     2581 cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat
     2641 gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt
     2701 gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga
     2761 gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat
     2821 cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc
     2881 ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt
     2941 taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg
     3001 gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac
     3061 cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacggcg gatcaggagg
     3121 ctctggcggt tcaggtggat caggcggtag cggaggttca ggtggtatgg acaagaagta
     3181 cagcatcggc ctggccatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta
     3241 caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa
     3301 gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa
     3361 gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat
     3421 cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt
     3481 cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga
     3541 ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag
     3601 caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg
     3661 gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt
     3721 catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg
     3781 cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct
     3841 gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggaaacctga ttgccctgag
     3901 cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca
     3961 gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca
     4021 gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat
     4081 cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata
     4141 cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga
     4201 gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg
     4261 cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg
     4321 caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt
     4381 cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg
     4441 gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac
     4501 cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat
     4561 gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg
     4621 cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga
     4681 gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac
     4741 caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa
     4801 aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa
     4861 agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga
     4921 tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga
     4981 cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact
     5041 gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga
     5101 caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa
     5161 gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc
     5221 cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa
     5281 agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc
     5341 caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga
     5401 cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag
     5461 agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga
     5521 agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca
     5581 gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga
     5641 ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gaccatatcg tgcctcagag
     5701 ctttctgaag gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg
     5761 caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca
     5821 gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag
     5881 aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg
     5941 gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga
     6001 gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga
     6061 tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca
     6121 cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga
     6181 aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag
     6241 cgagcagtct ggcggctcaa aaagaaccgc cgacggcagc gaattcgagc ccaagaagaa
     6301 gaggaaagtc taaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc
     6361 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     6421 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     6481 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     6541 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     6601 gaaagaacca gctggggctc gataccgtcg acctctagct agagcttggc gtaatcatgg
     6661 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     6721 ggaagcataa agtgtaaagc ctagggtgcc taatgagtga gctaactcac attaattgcg
     6781 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     6841 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     6901 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6961 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     7021 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     7081 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     7141 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     7201 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     7261 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     7321 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     7381 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     7441 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     7501 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     7561 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7621 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     7681 gacactcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7741 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7801 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7861 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7921 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7981 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     8041 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     8101 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     8161 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     8221 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     8281 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     8341 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     8401 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     8461 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     8521 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     8581 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     8641 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     8701 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     8761 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     8821 ggatcgggag atcgatctcc cgatccccta gggtcgattc tcagtacaat ctgc
//
