LOCUS       sequence-231809-        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: CP1041-ABEmax
FEATURES             Location/Qualifiers
     source          1..8871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        188..567
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        568..771
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     722..742
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     813..832
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        813..831
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             881..901
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3134..4849
                     /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             6293..6313
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6322..6339
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6362..6379)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6368..6592
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6663..6679)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6663..6679)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6676..6698)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6687..6703
                     /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(6711..6717,6718..6735,6736..6741))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6756..6777
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6894..6911)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7065..7653)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7145..7164)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7824..8615,8616..8684))
                     /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     8447..8466
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8685..8789)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(8868..8871,1..16))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
       61 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
      121 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
      181 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
      241 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      301 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      361 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      421 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      481 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      541 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      601 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      661 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      721 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
      781 aaccgtcaga tccgctagag atccgcggcc gctaatacga ctcactatag ggagagccgc
      841 caccatgaaa cggacagccg acggaagcga gttcgagtca ccaaagaaga agcggaaagt
      901 ctctgaagtc gagtttagcc acgagtattg gatgaggcac gcactgaccc tggcaaagcg
      961 agcatgggat gaaagagaag tccccgtggg cgccgtgctg gtgcacaaca atagagtgat
     1021 cggagaggga tggaacaggc caatcggccg ccacgaccct accgcacacg cagagatcat
     1081 ggcactgagg cagggaggcc tggtcatgca gaattaccgc ctgatcgatg ccaccctgta
     1141 tgtgacactg gagccatgcg tgatgtgcgc aggagcaatg atccacagca ggatcggaag
     1201 agtggtgttc ggagcacggg acgccaagac cggcgcagca ggctccctga tggatgtgct
     1261 gcaccacccc ggcatgaacc accgggtgga gatcacagag ggaatcctgg cagacgagtg
     1321 cgccgccctg ctgagcgatt tctttagaat gcggagacag gagatcaagg cccagaagaa
     1381 ggcacagagc tccaccgact ctggaggatc tagcggagga tcctctggaa gcgagacacc
     1441 aggcacaagc gagtccgcca caccagagag ctccggcggc tcctccggag gatcctctga
     1501 ggtggagttt tcccacgagt actggatgag acatgccctg accctggcca agagggcacg
     1561 cgatgagagg gaggtgcctg tgggagccgt gctggtgctg aacaatagag tgatcggcga
     1621 gggctggaac agagccatcg gcctgcacga cccaacagcc catgccgaaa ttatggccct
     1681 gagacagggc ggcctggtca tgcagaacta cagactgatt gacgccaccc tgtacgtgac
     1741 attcgagcct tgcgtgatgt gcgccggcgc catgatccac tctaggatcg gccgcgtggt
     1801 gtttggcgtg aggaacgcaa aaaccggcgc cgcaggctcc ctgatggacg tgctgcacta
     1861 ccccggcatg aatcaccgcg tcgaaattac cgagggaatc ctggcagatg aatgtgccgc
     1921 cctgctgtgc tatttctttc ggatgcctag acaggtgttc aatgctcaga agaaggccca
     1981 gagctccacc gactccggag gatctagcgg aggctcctct ggctctgaga cacctggcac
     2041 aagcgagagc gcaacacctg aaagcagcgg gggcagcagc ggggggtcaa acatcatgaa
     2101 ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga
     2161 gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg
     2221 gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg
     2281 cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa
     2341 ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct
     2401 ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct
     2461 ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc
     2521 caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt
     2581 cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa
     2641 cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa
     2701 gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca
     2761 ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga
     2821 cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga
     2881 gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt
     2941 caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga
     3001 cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca
     3061 gctgggaggc gacggcggat caggaggctc tggcggttca ggtggatcag gcggtagcgg
     3121 aggttcaggt ggtgacaaga agtacagcat cggcctggcc atcggcacca actctgtggg
     3181 ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa
     3241 caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga
     3301 aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa
     3361 ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt
     3421 cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc
     3481 catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca
     3541 cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc
     3601 cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga
     3661 caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga
     3721 ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag
     3781 caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct
     3841 gttcggaaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga
     3901 cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa
     3961 cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc
     4021 cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct
     4081 gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc
     4141 tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa
     4201 cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa
     4261 gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga
     4321 cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg
     4381 agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg
     4441 ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag
     4501 gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa
     4561 cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa
     4621 cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta
     4681 cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc
     4741 cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg
     4801 gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc
     4861 cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct
     4921 gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga
     4981 agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa
     5041 aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac
     5101 cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa
     5161 gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat
     5221 ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg
     5281 cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat
     5341 cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga
     5401 gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag
     5461 ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa
     5521 agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca
     5581 gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga
     5641 tgtggaccat atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct
     5701 gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa
     5761 gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt
     5821 cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat
     5881 caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc
     5941 ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac
     6001 cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga
     6061 gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct
     6121 gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga
     6181 cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt
     6241 cttctacagc tctggcggct caaaaagaac cgccgacggc agcgaattcg agcccaagaa
     6301 gaagaggaaa gtctaaccgg tcatcatcac catcaccatt gagtttaaac ccgctgatca
     6361 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     6421 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     6481 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     6541 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     6601 gcggaaataa ccagctgggg ctcgataccg tcgacctcta gctagagctt ggcgtaatca
     6661 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     6721 gccggaagca taaagtgtaa agcctagggt gcctaatgag tgagctaact cacattaatt
     6781 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     6841 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6901 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6961 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     7021 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     7081 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     7141 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     7201 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     7261 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     7321 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     7381 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     7441 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7501 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7561 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7621 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7681 tctgacactc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7741 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7801 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     7861 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     7921 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7981 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     8041 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     8101 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     8161 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     8221 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     8281 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     8341 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     8401 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     8461 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     8521 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     8581 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     8641 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     8701 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     8761 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     8821 gacggatcgg gagatcgatc tcccgatccc ctagggtcga ctctcagtac a
//
