LOCUS       sequence-256905-        7869 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 7869)
  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 7869)
  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: SaKKH-ABEmax
FEATURES             Location/Qualifiers
     source          1..7869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        103..482
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        483..686
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     637..657
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     728..747
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        728..746
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             796..816
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5206..5226
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5235..5252
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5275..5292)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5281..5505
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5576..5592)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5576..5592)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5589..5611)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5600..5616
                     /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(5624..5630,5631..5648,5649..5654))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5669..5690
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5807..5824)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5978..6566)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6058..6077)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6737..7528,7529..7597))
                     /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     7360..7379
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7598..7702)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7781..7800)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
       61 gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta
      121 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      181 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      241 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      301 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      361 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      421 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      481 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      541 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      601 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      661 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagagatccg
      721 cggccgctaa tacgactcac tatagggaga gccgccacca tgaaacggac agccgacgga
      781 agcgagttcg agtcaccaaa gaagaagcgg aaagtctctg aagtcgagtt tagccacgag
      841 tattggatga ggcacgcact gaccctggca aagcgagcat gggatgaaag agaagtcccc
      901 gtgggcgccg tgctggtgca caacaataga gtgatcggag agggatggaa caggccaatc
      961 ggccgccacg accctaccgc acacgcagag atcatggcac tgaggcaggg aggcctggtc
     1021 atgcagaatt accgcctgat cgatgccacc ctgtatgtga cactggagcc atgcgtgatg
     1081 tgcgcaggag caatgatcca cagcaggatc ggaagagtgg tgttcggagc acgggacgcc
     1141 aagaccggcg cagcaggctc cctgatggat gtgctgcacc accccggcat gaaccaccgg
     1201 gtggagatca cagagggaat cctggcagac gagtgcgccg ccctgctgag cgatttcttt
     1261 agaatgcgga gacaggagat caaggcccag aagaaggcac agagctccac cgactctgga
     1321 ggatctagcg gaggatcctc tggaagcgag acaccaggca caagcgagtc cgccacacca
     1381 gagagctccg gcggctcctc cggaggatcc tctgaggtgg agttttccca cgagtactgg
     1441 atgagacatg ccctgaccct ggccaagagg gcacgcgatg agagggaggt gcctgtggga
     1501 gccgtgctgg tgctgaacaa tagagtgatc ggcgagggct ggaacagagc catcggcctg
     1561 cacgacccaa cagcccatgc cgaaattatg gccctgagac agggcggcct ggtcatgcag
     1621 aactacagac tgattgacgc caccctgtac gtgacattcg agccttgcgt gatgtgcgcc
     1681 ggcgccatga tccactctag gatcggccgc gtggtgtttg gcgtgaggaa cgcaaaaacc
     1741 ggcgccgcag gctccctgat ggacgtgctg cactaccccg gcatgaatca ccgcgtcgaa
     1801 attaccgagg gaatcctggc agatgaatgt gccgccctgc tgtgctattt ctttcggatg
     1861 cctagacagg tgttcaatgc tcagaagaag gcccagagct ccaccgactc cggaggatct
     1921 agcggaggct cctctggctc tgagacacct ggcacaagcg agagcgcaac acctgaaagc
     1981 agcgggggca gcagcggggg gtcagggaag cgaaattaca ttctggggct ggccattggc
     2041 attacatcag tgggctatgg catcattgac tacgagacaa gggacgtgat cgacgccggc
     2101 gtgagactgt tcaaggaggc caacgtggag aacaatgagg gccggagatc caagagggga
     2161 gcaaggcgcc tgaagcggag aaggcgccac agaatccaga gagtgaagaa gctgctgttc
     2221 gattacaacc tgctgaccga ccactccgag ctgtctggca tcaatcctta tgaggccaga
     2281 gtgaagggcc tgtcccagaa gctgtctgag gaggagttta gcgccgccct gctgcacctg
     2341 gcaaagagga gaggcgtgca caacgtgaat gaggtggagg aggacaccgg caacgagctg
     2401 tccacaaagg agcagatcag ccgcaattcc aaggccctgg aggagaagta tgtggccgag
     2461 ctgcagctgg agcggctgaa gaaggatggc gaggtgaggg gctccatcaa tcgcttcaag
     2521 acctctgact acgtgaagga ggccaagcag ctgctgaagg tgcagaaggc ctaccaccag
     2581 ctggatcagt cctttatcga tacatatatc gacctgctgg agacaaggcg cacatactat
     2641 gagggaccag gagagggctc tcccttcggc tggaaggaca tcaaggagtg gtacgagatg
     2701 ctgatgggcc actgcaccta ttttccagag gagctgagaa gcgtgaagta cgcctataac
     2761 gccgatctgt acaacgccct gaatgacctg aacaacctgg tcatcaccag ggatgagaac
     2821 gagaagctgg agtactatga gaagttccag atcatcgaga acgtgttcaa gcagaagaag
     2881 aagcctacac tgaagcagat cgccaaggag atcctggtga acgaggagga catcaagggc
     2941 taccgcgtga cctccacagg caagccagag ttcaccaatc tgaaggtgta tcacgatatc
     3001 aaggacatca cagcccggaa ggagatcatc gagaacgccg agctgctgga tcagatcgcc
     3061 aagatcctga ccatctatca gagctccgag gacatccagg aggagctgac caacctgaat
     3121 agcgagctga cacaggagga gatcgagcag atcagcaatc tgaagggcta caccggcaca
     3181 cacaacctga gcctgaaggc catcaatctg atcctggatg agctgtggca cacaaacgac
     3241 aatcagatcg ccatctttaa ccggctgaag ctggtgccaa agaaggtgga cctgtcccag
     3301 cagaaggaga tcccaaccac actggtggac gatttcatcc tgtctcccgt ggtgaagcgg
     3361 agcttcatcc agagcatcaa agtgatcaac gccatcatca agaagtacgg cctgcccaat
     3421 gatatcatca tcgagctggc cagggagaag aactccaagg acgcccagaa gatgatcaat
     3481 gagatgcaga agaggaaccg ccagaccaat gagcggatcg aggagatcat cagaaccaca
     3541 ggcaaggaga acgccaagta cctgatcgag aagatcaagc tgcacgatat gcaggagggc
     3601 aagtgtctgt attctctgga ggccatccct ctggaggacc tgctgaacaa tccattcaac
     3661 tacgaggtgg atcacatcat cccccggagc gtgagcttcg acaattcttt taacaataag
     3721 gtgctggtga agcaggagga gaacagcaag aagggcaata ggaccccttt ccagtacctg
     3781 tctagctccg attctaagat cagctacgag acattcaaga agcacatcct gaatctggcc
     3841 aagggcaagg gccgcatcag caagaccaag aaggagtacc tgctggagga gcgggacatc
     3901 aacagattct ccgtgcagaa ggacttcatc aaccggaatc tggtggacac cagatacgcc
     3961 acacgcggcc tgatgaatct gctgcggtct tatttcagag tgaacaatct ggatgtgaag
     4021 gtgaagagca tcaacggcgg cttcacctcc tttctgcgga gaaagtggaa gtttaagaag
     4081 gagcgcaaca agggctataa gcaccacgcc gaggatgccc tgatcatcgc caatgccgac
     4141 ttcatcttta aggagtggaa gaagctggac aaggccaaga aagtgatgga gaaccagatg
     4201 ttcgaggaga agcaggccga gagcatgccc gagatcgaga cagagcagga gtacaaggag
     4261 attttcatca cacctcacca gatcaagcac atcaaggact tcaaggacta caagtattct
     4321 cacagggtgg ataagaagcc caaccgcaag ctgatcaatg acaccctgta tagcacacgg
     4381 aaggacgata agggcaatac cctgatcgtg aacaatctga acggcctgta cgacaaggat
     4441 aatgacaagc tgaagaagct gatcaacaag tctcccgaga agctgctgat gtaccaccac
     4501 gatcctcaga catatcagaa gctgaagctg atcatggagc agtacggcga cgagaagaac
     4561 ccactgtata agtactatga ggagacaggc aactacctga caaagtatag caagaaggat
     4621 aatggccccg tgatcaagaa gatcaagtac tatggcaaca agctgaatgc ccacctggac
     4681 atcaccgacg attaccctaa ctctcgcaat aaggtggtga agctgagcct gaagccatac
     4741 cggttcgacg tgtacctgga caacggcgtg tataagtttg tgacagtgaa gaatctggat
     4801 gtgatcaaga aggagaacta ctatgaggtg aacagcaagt gctacgagga ggccaagaag
     4861 ctgaagaaga tcagcaacca ggccgagttc atcgcctctt tttacaagaa tgacctgatc
     4921 aagatcaatg gcgagctgta tagagtgatc ggcgtgaaca atgatctgct gaacagaatc
     4981 gaagtgaata tgatcgacat cacctacagg gagtatctgg agaacatgaa tgataagagg
     5041 ccccctcata tcatcaagac catcgcctct aagacacaga gcatcaagaa gtacagcaca
     5101 gacatcctgg ggaacctgta tgaagtcaag agcaagaaac atcctcagat tatcaagaaa
     5161 ggctctggcg gctcaaaaag aaccgccgac ggcagcgaat tcgagcccaa gaagaagagg
     5221 aaagtctaac cggtcatcat caccatcacc attgagttta aacccgctga tcagcctcga
     5281 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     5341 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     5401 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     5461 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     5521 gaaccagctg gggctcgata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     5581 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5641 gcataaagtg taaagcctag ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5701 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5761 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5821 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5881 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5941 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     6001 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6061 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6121 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6181 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6241 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6301 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6361 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6421 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6481 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6541 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgaca
     6601 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6661 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6721 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6781 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6841 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6901 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6961 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     7021 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7081 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7141 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7201 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7261 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7321 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7381 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7441 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7501 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7561 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7621 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7681 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     7741 cgggagatcg atctcccgat cccctagggt cgactctcag tacaatctgc tctgatgccg
     7801 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     7861 gcaaaattt
//
