LOCUS       sequence-231817-        8871 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 8871)
  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 8871)
  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: CP1300-ABEmax
FEATURES             Location/Qualifiers
     source          1..8871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        174..553
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        554..757
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     708..728
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     799..818
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        799..817
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             867..887
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2343..4058
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
     CDS             6279..6299
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6308..6325
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6348..6365)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6354..6578
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6649..6665)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6649..6665)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6662..6684)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6673..6689
                     /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(6697..6703,6704..6721,6722..6727))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6742..6763
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6880..6897)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7051..7639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7131..7150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7810..8601,8602..8670))
                     /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     8433..8452
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8671..8775)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(8854..8871,1..2))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
       61 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
      121 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
      181 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
      241 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
      301 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      361 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      421 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      481 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      541 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      601 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      661 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      721 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg
      781 ctagagatcc gcggccgcta atacgactca ctatagggag agccgccacc atgaaacgga
      841 cagccgacgg aagcgagttc gagtcaccaa agaagaagcg gaaagtctct gaagtcgagt
      901 ttagccacga gtattggatg aggcacgcac tgaccctggc aaagcgagca tgggatgaaa
      961 gagaagtccc cgtgggcgcc gtgctggtgc acaacaatag agtgatcgga gagggatgga
     1021 acaggccaat cggccgccac gaccctaccg cacacgcaga gatcatggca ctgaggcagg
     1081 gaggcctggt catgcagaat taccgcctga tcgatgccac cctgtatgtg acactggagc
     1141 catgcgtgat gtgcgcagga gcaatgatcc acagcaggat cggaagagtg gtgttcggag
     1201 cacgggacgc caagaccggc gcagcaggct ccctgatgga tgtgctgcac caccccggca
     1261 tgaaccaccg ggtggagatc acagagggaa tcctggcaga cgagtgcgcc gccctgctga
     1321 gcgatttctt tagaatgcgg agacaggaga tcaaggccca gaagaaggca cagagctcca
     1381 ccgactctgg aggatctagc ggaggatcct ctggaagcga gacaccaggc acaagcgagt
     1441 ccgccacacc agagagctcc ggcggctcct ccggaggatc ctctgaggtg gagttttccc
     1501 acgagtactg gatgagacat gccctgaccc tggccaagag ggcacgcgat gagagggagg
     1561 tgcctgtggg agccgtgctg gtgctgaaca atagagtgat cggcgagggc tggaacagag
     1621 ccatcggcct gcacgaccca acagcccatg ccgaaattat ggccctgaga cagggcggcc
     1681 tggtcatgca gaactacaga ctgattgacg ccaccctgta cgtgacattc gagccttgcg
     1741 tgatgtgcgc cggcgccatg atccactcta ggatcggccg cgtggtgttt ggcgtgagga
     1801 acgcaaaaac cggcgccgca ggctccctga tggacgtgct gcactacccc ggcatgaatc
     1861 accgcgtcga aattaccgag ggaatcctgg cagatgaatg tgccgccctg ctgtgctatt
     1921 tctttcggat gcctagacag gtgttcaatg ctcagaagaa ggcccagagc tccaccgact
     1981 ccggaggatc tagcggaggc tcctctggct ctgagacacc tggcacaagc gagagcgcaa
     2041 cacctgaaag cagcgggggc agcagcgggg ggtcaaagcc catcagagag caggccgaga
     2101 atatcatcca cctgtttacc ctgaccaatc tgggagcccc tgccgccttc aagtactttg
     2161 acaccaccat cgaccggaag aggtacacca gcaccaaaga ggtgctggac gccaccctga
     2221 tccaccagag catcaccggc ctgtacgaga cacggatcga cctgtctcag ctgggaggcg
     2281 acggcggatc aggaggctct ggcggttcag gtggatcagg cggtagcgga ggttcaggtg
     2341 gtgacaagaa gtacagcatc ggcctggcca tcggcaccaa ctctgtgggc tgggccgtga
     2401 tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc
     2461 acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg
     2521 ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct
     2581 atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac
     2641 tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca
     2701 acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga
     2761 aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca
     2821 tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg
     2881 tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca
     2941 tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac
     3001 ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc
     3061 tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg
     3121 atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc
     3181 agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc
     3241 tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta
     3301 tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc
     3361 agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg
     3421 gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg
     3481 aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga
     3541 agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg
     3601 ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg
     3661 agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca
     3721 gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag
     3781 tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga
     3841 acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt
     3901 ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga
     3961 gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg
     4021 tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct
     4081 ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta
     4141 tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc
     4201 tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc
     4261 acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca
     4321 ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg
     4381 atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca
     4441 gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc
     4501 acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag
     4561 tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga
     4621 tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa
     4681 tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg
     4741 tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg
     4801 atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata
     4861 tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg
     4921 acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga
     4981 actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga
     5041 ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc
     5101 tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca
     5161 ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca
     5221 agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact
     5281 accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt
     5341 accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga
     5401 tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca
     5461 acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc
     5521 ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg
     5581 ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc
     5641 agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg
     5701 ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct
     5761 attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga
     5821 aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact
     5881 ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt
     5941 actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc
     6001 agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc
     6061 actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac
     6121 agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga
     6181 tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggattctg
     6241 gcggctcaaa aagaaccgcc gacggcagcg aattcgagcc caagaagaag aggaaagtct
     6301 aaccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc
     6361 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     6421 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     6481 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     6541 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     6601 ctggggctcg ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     6661 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6721 gtgtaaagcc tagggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6781 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6841 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6901 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6961 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     7021 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     7081 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     7141 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     7201 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     7261 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     7321 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     7381 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7441 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     7501 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7561 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7621 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acactcagtg
     7681 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7741 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7801 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7861 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7921 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7981 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     8041 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     8101 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     8161 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     8221 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     8281 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     8341 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     8401 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8461 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8521 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8581 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8641 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8701 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8761 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     8821 tcgatctccc gatcccctag ggtcgactct cagtacaatc tgctctgatg c
//
