LOCUS       sequence-304588-        7275 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  A-to-G base editor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
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: SaABE8e(TadA V106W)
FEATURES             Location/Qualifiers
     source          1..7275
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    520..541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(556..561,562..579,580..586)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    594..610
                     /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)."
     primer_bind     599..621
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     618..634
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     618..634
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    705..929
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     918..935
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(958..975)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(984..1004)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(4800..4820)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4869..4888)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4870..4888)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4930..5133)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4959..4979)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5134..5513
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5685..5704
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5783..5887
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5888..5956,5957..6748)
                     /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     complement(6106..6125)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
      541 ttaggcaccc taggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
      601 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc tctagctaga
      661 ggtcgacggt atcgagcccc agctggttct ttccgcctca gaagccatag agcccaccgc
      721 atccccagca tgcctgctat tgtcttccca atcctccccc ttgctgtcct gccccacccc
      781 accccccaga atagaatgac acctactcag acaatgcgat gcaatttcct cattttatta
      841 ggaaaggaca gtgggagtgg caccttccag ggtcaaggaa ggcacggggg aggggcaaac
      901 aacagatggc tggcaactag aaggcacagt cgaggctgat cagcgggttt aaactcaatg
      961 gtgatggtga tgatgaccgg ttagactttc ctcttcttct tgggctcgaa ttcgctgccg
     1021 tcggcggttc tttttgagcc gccagagcct ttcttgataa tctgaggatg tttcttgctc
     1081 ttgacttcat acaggttccc caggatgtct gtgctgtact tcttgatgct ctgtgtctta
     1141 gaggcgatgg tcttgatgat gcgagggggc ctcttatcat tcatgttctc cagatactcc
     1201 ctgtaggtga tgtcgatcat attcacttcg attctgttca gcagatcatt gttcacgccg
     1261 atcactctat acagctcgcc attgatcttg atcaggtcat tgttgtaaaa agaggcgatg
     1321 aactcggcct ggttgctgat cttcttcagc ttcttggcct cctcgtagca cttgctgttc
     1381 acctcatagt agttctcctt cttgatcaca tccagattct tcactgtcac aaacttatac
     1441 acgccgttgt ccaggtacac gtcgaaccgg tatggcttca ggctcagctt caccacctta
     1501 ttgcgagagt tagggtaatc gtcggtgatg tccaggtggg cattcagctt gttgccatag
     1561 tacttgatct tcttgatcac ggggccatta tccttcttgc tatactttgt caggtagttg
     1621 cctgtctcct catagtactt atacagtggg ttcttctcgt cgccgtactg ctccatgatc
     1681 agcttcagct tctgatatgt ctgaggatcg tggtggtaca tcagcagctt ctcgggagac
     1741 ttgttgatca gcttcttcag cttgtcatta tccttgtcgt acaggccgtt cagattgttc
     1801 acgatcaggg tattgccctt atcgtccttc cgtgtgctat acagggtgtc attgatcagc
     1861 tcgcggttgg gcttcttatc caccctgtga gaatacttgt agtccttgaa gtccttgatg
     1921 tgcttgatct ggtgaggtgt gatgaaaatc tccttgtact cctgctctgt ctcgatctcg
     1981 ggcatgctct cggcctgctt ctcctcgaac atctggttct ccatcacttt cttggccttg
     2041 tccagcttct tccactcctt aaagatgaag tcggcattgg cgatgatcag ggcatcctcg
     2101 gcgtggtgct tatagccctt gttgcgctcc ttcttaaact tccactttct ccgcagaaag
     2161 gaggtgaagc cgccgttgat gctcttcacc ttcacatcca gattgttcac tctgaaataa
     2221 gaccgcagca gattcatcag gccgcgtgtg gcgtatctgg tgtccaccag attccggttg
     2281 atgaagtcct tctgcacgga gaatctgttg atgtcccgct cctccagcag gtactccttc
     2341 ttggtcttgc tgatgcggcc cttgcccttg gccagattca ggatgtgctt cttgaatgtc
     2401 tcgtagctga tcttagaatc ggagctagac aggtactgga aaggggtcct attgcccttc
     2461 ttgctgttct cctcctgctt caccagcacc ttattgttaa aagaattgtc gaagctcacg
     2521 ctccggggga tgatgtgatc cacctcgtag ttgaatggat tgttcagcag gtcctccaga
     2581 gggatggcct ccagagaata cagacacttg ccctcctgca tatcgtgcag cttgatcttc
     2641 tcgatcaggt acttggcgtt ctccttgcct gtggttctga tgatctcctc gatccgctca
     2701 ttggtctggc ggttcctctt ctgcatctca ttgatcatct tctgggcgtc cttggagttc
     2761 ttctccctgg ccagctcgat gatgatatca ttgggcaggc cgtacttctt gatgatggcg
     2821 ttgatcactt tgatgctctg gatgaagctc cgcttcacca cgggagacag gatgaaatcg
     2881 tccaccagtg tggttgggat ctccttctgc tgggacaggt ccaccttctt tggcaccagc
     2941 ttcagccggt taaagatggc gatctgattg tcgtttgtgt gccacagctc atccaggatc
     3001 agattgatgg ccttcaggct caggttgtgt gtgccggtgt agcccttcag attgctgatc
     3061 tgctcgatct cctcctgtgt cagctcgcta ttcaggttgg tcagctcctc ctggatgtcc
     3121 tcggagctct gatagatggt caggatcttg gcgatctgat ccagcagctc ggcgttctcg
     3181 atgatctcct tccgggctgt gatgtccttg atatcgtgat acaccttcag attggtgaac
     3241 tctggcttgc ctgtggaggt cacgcggtag cccttgatgt cctcctcgtt caccaggatc
     3301 tccttggcga tctgcttcag tgtaggcttc ttcttctgct tgaacacgtt ctcgatgatc
     3361 tggaacttct catagtactc cagcttctcg ttctcatccc tggtgatgac caggttgttc
     3421 aggtcattca gggcgttgta cagatcggcg ttataggcgt acttcacgct tctcagctcc
     3481 tctggaaaat aggtgcagtg gcccatcagc atctcgtacc actccttgat gtccttccag
     3541 ccgaagggag agccctctcc tggtccctca tagtatgtgc gccttgtctc cagcaggtcg
     3601 atatatgtat cgataaagga ctgatccagc tggtggtagg ccttctgcac cttcagcagc
     3661 tgcttggcct ccttcacgta gtcagaggtc ttgaagcgat tgatggagcc cctcacctcg
     3721 ccatccttct tcagccgctc cagctgcagc tcggccacat acttctcctc cagggccttg
     3781 gaattgcggc tgatctgctc ctttgtggac agctcgttgc cggtgtcctc ctccacctca
     3841 ttcacgttgt gcacgcctct cctctttgcc aggtgcagca gggcggcgct aaactcctcc
     3901 tcagacagct tctgggacag gcccttcact ctggcctcat aaggattgat gccagacagc
     3961 tcggagtggt cggtcagcag gttgtaatcg aacagcagct tcttcactct ctggattctg
     4021 tggcgccttc tccgcttcag gcgccttgct cccctcttgg atctccggcc ctcattgttc
     4081 tccacgttgg cctccttgaa cagtctcacg ccggcgtcga tcacgtccct tgtctcgtag
     4141 tcaatgatgc catagcccac tgatgtaatg ccaatggcca gccccagaat gtaatttcgc
     4201 ttccctgacc ccccgctgct gcccccgctg ctttcaggtg ttgcgctctc gcttgtgcca
     4261 ggtgtctcag agccagagga gcctccgcta gatcctccgg agttgatgga gctctgggcc
     4321 ttcttctgag cattgaacac ctgtctaggc atccgataga aatcgcacag cagggcggca
     4381 cattcatctg ccaggattcc ctcggtaatt tcgacgcggt gattcatgcc ggggtagttc
     4441 agcacgttca tcagggagcc tgcggcgcct cttttagaat ttctccatcc aaacaccacg
     4501 cggccgatcc tagagtggat catggcgccg gcgcacatca cgcaaggctc gaatgtcacg
     4561 tacagggtgg cgtcaatcag tctgtagttc tgcatgacca ggccgccctg tctcagggcc
     4621 ataatttcgg catgggctgt tgggtcgtgc aggccgatgg ctctgttcca gccctcgccg
     4681 atcactctat tgttcagcac cagcacggct cccacaggca cctccctctc atcccgtgcc
     4741 ctcttggcca gggtcagggc atgtctcatc cagtactcgt gggaaaactc cacctcagag
     4801 actttccgct tcttctttgg tgactcgaac tcgcttccgt cggctgtccg tttcatggtg
     4861 gcggctctcc ctatagtgag tcgtattagc ggccgcggat ctctagcgga tctgacggtt
     4921 cactaaacca gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt
     4981 caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa
     5041 actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc
     5101 acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta
     5161 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     5221 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     5281 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     5341 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     5401 tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactcaatat atgggctatg
     5461 aactaatgac cccgtaattg attactatta ataactagtc aataatcaat gtcaacgcgt
     5521 atatctggcc cgtacatcgc gaagcagcgc aaaacgccta accctaagca gattcttcat
     5581 gcaattgtcg gtcaagcctt gccttgttgt agcttaaatt ttgctcgcgc actactcagc
     5641 gacctccaac acacaagcag ggagcagata ctggcttaac tatgcggcat cagagcagat
     5701 tgtactgaga gtcgacccta ggggatcggg agatcgatct cccgatccgt cgacgtcagg
     5761 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5821 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5881 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5941 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     6001 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     6061 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     6121 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     6181 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     6241 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     6301 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     6361 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     6421 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     6481 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6541 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6601 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6661 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6721 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6781 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6841 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagtgtcag accccgtaga
     6901 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6961 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     7021 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     7081 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7141 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7201 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     7261 cagcttggag cgaac
//
