LOCUS       sequence-369345-        8271 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 8271)
  AUTHORS   Cao X, Guo J, Huang S, Yu W, Li G, An L, Li X, Tao W, Liu Q, Huang 
            X, Jin X, Ma X
  TITLE     Engineering of near-PAMless adenine base editor with enhanced 
            editing activity and reduced off-target.
  JOURNAL   Mol Ther Nucleic Acids. 2022 May 4;28:732-742. doi: 
            10.1016/j.omtn.2022.04.032. eCollection 2022 Jun 14.
  PUBMED    35664696
REFERENCE   2  (bases 1 to 8271)
  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: "Mol Ther 
            Nucleic Acids."; date: "2022-05-4"; volume: "28:732-742. doi"; 
            pages: " 10.1016/j.omtn.2022.04.032. eCollection 2022 Jun 14"
COMMENT     Sequence Label: CE-8e-SpRY
FEATURES             Location/Qualifiers
     source          1..8271
                     /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             472..2187
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAERTRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
     CDS             5257..5277
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5286..5303
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5326..5343)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5332..5556
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5627..5643)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5627..5643)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5640..5662)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5651..5667
                     /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(5675..5681,5682..5699,5700..5705))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5720..5741
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5858..5875)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6029..6617)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6109..6128)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6788..7579,7580..7648))
                     /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     7411..7430
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7649..7753)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7832..7851)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(8023..8271,1..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 aagtcagcag tgacaagaag
      481 tacagcatcg gcctggccat cggcaccaac tctgtgggct gggccgtgat caccgacgag
      541 tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca cagcatcaag
      601 aagaacctga tcggagccct gctgttcgac agcggcgaaa cagccgagag aacccggctg
      661 aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta tctgcaagag
      721 atcttcagca acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc
      781 ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac
      841 gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac
      901 agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc
      961 cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg
     1021 ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc
     1081 ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat
     1141 ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggaaacct gattgccctg
     1201 agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg
     1261 cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac
     1321 cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac
     1381 atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga
     1441 tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct
     1501 gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacattgac
     1561 ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac
     1621 ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc
     1681 ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg
     1741 cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg
     1801 accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg
     1861 atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag
     1921 ggcgcttccg cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac
     1981 gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta taacgagctg
     2041 accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag
     2101 aaaaaggcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg
     2161 aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa
     2221 gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat caaggacaag
     2281 gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct gaccctgaca
     2341 ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca cctgttcgac
     2401 gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag gctgagccgg
     2461 aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga tttcctgaag
     2521 tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag cctgaccttt
     2581 aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca cgagcacatt
     2641 gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt gaaggtggtg
     2701 gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat cgaaatggcc
     2761 agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat gaagcggatc
     2821 gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt ggaaaacacc
     2881 cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga tatgtacgtg
     2941 gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggaccatat cgtgcctcag
     3001 agctttctga aggacgactc catcgacaac aaggtgctga ccagaagcga caagaaccgg
     3061 ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg
     3121 cagctgctga acgccaagct gattacccag agaaagttcg acaatctgac caaggccgag
     3181 agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct ggtggaaacc
     3241 cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac taagtacgac
     3301 gagaatgaca agctgatccg ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc
     3361 gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc
     3421 cacgacgcct acctaaacgc cgtcgtggga accgccctga tcaaaaagta ccctaagctg
     3481 gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat gatcgccaag
     3541 agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa catcatgaac
     3601 tttttcaagt ctggaggatc tagcggtggt tcctctggaa gcgagacacc aggcacaagc
     3661 gagtccgcca caccagagag ctccggcggc tcctccggag gatcctctga ggtggagttt
     3721 tcccacgagt actggatgag acatgccctg accctggcca agagggcacg ggatgagagg
     3781 gaggtgcctg tgggagccgt gctggtgctg aacaatagag tgatcggcga gggctggaac
     3841 agagccatcg gcctgcacga cccaacagcc catgccgaaa ttatggccct gagacagggc
     3901 ggcctggtca tgcagaacta cagactgatt gacgccaccc tgtacgtgac attcgagcct
     3961 tgcgtgatgt gcgccggcgc catgatccac tctaggatcg gccgcgtggt gtttggcgtg
     4021 aggaactcaa aaagaggcgc cgcaggctcc ctgatgaacg tgctgaacta ccccggcatg
     4081 aatcaccgcg tcgaaattac cgagggaatc ctggcagatg aatgtgccgc cctgctgtgc
     4141 gatttctatc ggatgcctag acaggtgttc aatgctcaga agaaggccca gagctccatc
     4201 aactccggag gatctagcgg aggctcctct ggctctgaga cacctggcac aagcgagagc
     4261 gcaacacctg aaagcagcgg gggcagcagc ggggggtcag agacaaacgg cgaaaccggg
     4321 gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc
     4381 caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc
     4441 agacccaaga ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag
     4501 tacggcggct tcctgtggcc caccgtggcc tattctgtgc tggtggtggc caaagtggaa
     4561 aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa
     4621 agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg
     4681 aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg
     4741 aagagaatgc tggcctctgc caagcagctg cagaagggaa acgaactggc cctgccctcc
     4801 aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag
     4861 gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc
     4921 gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg
     4981 ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc
     5041 cacctgttta ccctgaccag actgggagcc cctagagcct tcaagtactt tgacaccacc
     5101 atcgacccca agcagtacag aagcaccaaa gaggtgctgg acgccaccct gatccaccag
     5161 agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg tgactctggc
     5221 ggctcaaaaa gaaccgccga cggcagcgaa ttcgagccca agaagaagag gaaagtctaa
     5281 ccggtcatca tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt
     5341 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     5401 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     5461 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     5521 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     5581 ggggctcgat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     5641 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5701 gtaaagccta gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5761 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5821 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5881 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5941 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6001 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6061 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6121 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6181 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6241 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6301 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6361 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6421 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6481 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6541 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6601 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac actcagtgga
     6661 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6721 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6781 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6841 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6901 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6961 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7021 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7081 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7141 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7201 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7261 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7321 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7381 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7441 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7501 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7561 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7621 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7681 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7741 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     7801 gatctcccga tcccctaggg tcgactctca gtacaatctg ctctgatgcc gcatagttaa
     7861 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     7921 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
     7981 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
     8041 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     8101 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     8161 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     8221 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat c
//
