LOCUS       sequence-272933-        7275 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 7275)
  AUTHORS   Richter MF, Zhao KT, Eton E, Lapinaite A, Newby GA, Thuronyi BW, 
            Wilson C, Koblan LW, Zeng J, Bauer DE, Doudna JA, Liu DR
  TITLE     Phage-assisted evolution of an adenine base editor with improved Cas
            domain compatibility and activity.
  JOURNAL   Nat Biotechnol. 2020 Jul;38(7):883-891. doi: 
            10.1038/s41587-020-0453-z. Epub 2020 Mar 16.
  PUBMED    32433547
REFERENCE   2  (bases 1 to 7275)
  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"; doi: 
            "10.1038/s41587-020-0453-z"; journalName: "Nat Biotechnol"; date: 
            "2020-07"; volume: "38"; issue: "7"; pages: "883-891"
COMMENT     Sequence Label: SaKKH-ABE8e
FEATURES             Location/Qualifiers
     source          1..7275
                     /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             4261..4281
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4290..4307
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4330..4347)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4336..4560
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4631..4647)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4631..4647)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4644..4666)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4655..4671
                     /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(4679..4685,4686..4703,4704..4709))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4724..4745
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4862..4879)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5033..5621)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5113..5132)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5792..6583,6584..6652))
                     /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     6415..6434
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6653..6757)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6836..6855)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(7027..7275,1..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 ggtggagttt
      481 tcccacgagt actggatgag acatgccctg accctggcca agagggcacg ggatgagagg
      541 gaggtgcctg tgggagccgt gctggtgctg aacaatagag tgatcggcga gggctggaac
      601 agagccatcg gcctgcacga cccaacagcc catgccgaaa ttatggccct gagacagggc
      661 ggcctggtca tgcagaacta cagactgatt gacgccaccc tgtacgtgac attcgagcct
      721 tgcgtgatgt gcgccggcgc catgatccac tctaggatcg gccgcgtggt gtttggcgtg
      781 aggaactcaa aaagaggcgc cgcaggctcc ctgatgaacg tgctgaacta ccccggcatg
      841 aatcaccgcg tcgaaattac cgagggaatc ctggcagatg aatgtgccgc cctgctgtgc
      901 gatttctatc ggatgcctag acaggtgttc aatgctcaga agaaggccca gagctccatc
      961 aactccggag gatctagcgg aggctcctct ggctctgaga cacctggcac aagcgagagc
     1021 gcaacacctg aaagcagcgg gggcagcagc ggggggtcag ggaagcgaaa ttacattctg
     1081 gggctggcca ttggcattac atcagtgggc tatggcatca ttgactacga gacaagggac
     1141 gtgatcgacg ccggcgtgag actgttcaag gaggccaacg tggagaacaa tgagggccgg
     1201 agatccaaga ggggagcaag gcgcctgaag cggagaaggc gccacagaat ccagagagtg
     1261 aagaagctgc tgttcgatta caacctgctg accgaccact ccgagctgtc tggcatcaat
     1321 ccttatgagg ccagagtgaa gggcctgtcc cagaagctgt ctgaggagga gtttagcgcc
     1381 gccctgctgc acctggcaaa gaggagaggc gtgcacaacg tgaatgaggt ggaggaggac
     1441 accggcaacg agctgtccac aaaggagcag atcagccgca attccaaggc cctggaggag
     1501 aagtatgtgg ccgagctgca gctggagcgg ctgaagaagg atggcgaggt gaggggctcc
     1561 atcaatcgct tcaagacctc tgactacgtg aaggaggcca agcagctgct gaaggtgcag
     1621 aaggcctacc accagctgga tcagtccttt atcgatacat atatcgacct gctggagaca
     1681 aggcgcacat actatgaggg accaggagag ggctctccct tcggctggaa ggacatcaag
     1741 gagtggtacg agatgctgat gggccactgc acctattttc cagaggagct gagaagcgtg
     1801 aagtacgcct ataacgccga tctgtacaac gccctgaatg acctgaacaa cctggtcatc
     1861 accagggatg agaacgagaa gctggagtac tatgagaagt tccagatcat cgagaacgtg
     1921 ttcaagcaga agaagaagcc tacactgaag cagatcgcca aggagatcct ggtgaacgag
     1981 gaggacatca agggctaccg cgtgacctcc acaggcaagc cagagttcac caatctgaag
     2041 gtgtatcacg atatcaagga catcacagcc cggaaggaga tcatcgagaa cgccgagctg
     2101 ctggatcaga tcgccaagat cctgaccatc tatcagagct ccgaggacat ccaggaggag
     2161 ctgaccaacc tgaatagcga gctgacacag gaggagatcg agcagatcag caatctgaag
     2221 ggctacaccg gcacacacaa cctgagcctg aaggccatca atctgatcct ggatgagctg
     2281 tggcacacaa acgacaatca gatcgccatc tttaaccggc tgaagctggt gccaaagaag
     2341 gtggacctgt cccagcagaa ggagatccca accacactgg tggacgattt catcctgtct
     2401 cccgtggtga agcggagctt catccagagc atcaaagtga tcaacgccat catcaagaag
     2461 tacggcctgc ccaatgatat catcatcgag ctggccaggg agaagaactc caaggacgcc
     2521 cagaagatga tcaatgagat gcagaagagg aaccgccaga ccaatgagcg gatcgaggag
     2581 atcatcagaa ccacaggcaa ggagaacgcc aagtacctga tcgagaagat caagctgcac
     2641 gatatgcagg agggcaagtg tctgtattct ctggaggcca tccctctgga ggacctgctg
     2701 aacaatccat tcaactacga ggtggatcac atcatccccc ggagcgtgag cttcgacaat
     2761 tcttttaaca ataaggtgct ggtgaagcag gaggagaaca gcaagaaggg caataggacc
     2821 cctttccagt acctgtctag ctccgattct aagatcagct acgagacatt caagaagcac
     2881 atcctgaatc tggccaaggg caagggccgc atcagcaaga ccaagaagga gtacctgctg
     2941 gaggagcggg acatcaacag attctccgtg cagaaggact tcatcaaccg gaatctggtg
     3001 gacaccagat acgccacacg cggcctgatg aatctgctgc ggtcttattt cagagtgaac
     3061 aatctggatg tgaaggtgaa gagcatcaac ggcggcttca cctcctttct gcggagaaag
     3121 tggaagttta agaaggagcg caacaagggc tataagcacc acgccgagga tgccctgatc
     3181 atcgccaatg ccgacttcat ctttaaggag tggaagaagc tggacaaggc caagaaagtg
     3241 atggagaacc agatgttcga ggagaagcag gccgagagca tgcccgagat cgagacagag
     3301 caggagtaca aggagatttt catcacacct caccagatca agcacatcaa ggacttcaag
     3361 gactacaagt attctcacag ggtggataag aagcccaacc gcaagctgat caatgacacc
     3421 ctgtatagca cacggaagga cgataagggc aataccctga tcgtgaacaa tctgaacggc
     3481 ctgtacgaca aggataatga caagctgaag aagctgatca acaagtctcc cgagaagctg
     3541 ctgatgtacc accacgatcc tcagacatat cagaagctga agctgatcat ggagcagtac
     3601 ggcgacgaga agaacccact gtataagtac tatgaggaga caggcaacta cctgacaaag
     3661 tatagcaaga aggataatgg ccccgtgatc aagaagatca agtactatgg caacaagctg
     3721 aatgcccacc tggacatcac cgacgattac cctaactctc gcaataaggt ggtgaagctg
     3781 agcctgaagc cataccggtt cgacgtgtac ctggacaacg gcgtgtataa gtttgtgaca
     3841 gtgaagaatc tggatgtgat caagaaggag aactactatg aggtgaacag caagtgctac
     3901 gaggaggcca agaagctgaa gaagatcagc aaccaggccg agttcatcgc ctctttttac
     3961 aagaatgacc tgatcaagat caatggcgag ctgtatagag tgatcggcgt gaacaatgat
     4021 ctgctgaaca gaatcgaagt gaatatgatc gacatcacct acagggagta tctggagaac
     4081 atgaatgata agaggccccc tcatatcatc aagaccatcg cctctaagac acagagcatc
     4141 aagaagtaca gcacagacat cctggggaac ctgtatgaag tcaagagcaa gaaacatcct
     4201 cagattatca agaaaggctc tggcggctca aaaagaaccg ccgacggcag cgaattcgag
     4261 cccaagaaga agaggaaagt ctaaccggtc atcatcacca tcaccattga gtttaaaccc
     4321 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     4381 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     4441 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     4501 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     4561 cttctgaggc ggaaagaacc agctggggct cgataccgtc gacctctagc tagagcttgg
     4621 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4681 acatacgagc cggaagcata aagtgtaaag cctagggtgc ctaatgagtg agctaactca
     4741 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4801 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4861 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4921 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4981 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5041 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5101 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5161 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5221 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5281 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5341 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5401 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5461 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5521 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5581 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5641 ctacggggtc tgacactcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5701 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5761 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5821 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5881 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5941 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6001 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6061 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6121 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6181 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6241 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6301 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6361 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6421 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6481 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6541 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6601 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6661 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6721 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6781 ctgacgtcga cggatcggga gatcgatctc ccgatcccct agggtcgact ctcagtacaa
     6841 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     6901 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     6961 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
     7021 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     7081 atagcccata tattgagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     7141 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     7201 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     7261 tacatcaagt gtatc
//
