LOCUS       sequence-231829-        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: SaABEmax
FEATURES             Location/Qualifiers
     source          1..7869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        118..497
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        498..701
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     652..672
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     743..762
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        743..761
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             811..831
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5221..5241
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5250..5267
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5290..5307)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5296..5520
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5591..5607)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5591..5607)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5604..5626)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5615..5631
                     /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(5639..5645,5646..5663,5664..5669))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5684..5705
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5822..5839)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5993..6581)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6073..6092)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6752..7543,7544..7612))
                     /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     7375..7394
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7613..7717)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7796..7815)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
       61 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      121 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      181 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      241 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      301 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      361 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      421 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      481 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      541 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      601 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      661 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
      721 tccgctagag atccgcggcc gctaatacga ctcactatag ggagagccgc caccatgaaa
      781 cggacagccg acggaagcga gttcgagtca ccaaagaaga agcggaaagt ctctgaagtc
      841 gagtttagcc acgagtattg gatgaggcac gcactgaccc tggcaaagcg agcatgggat
      901 gaaagagaag tccccgtggg cgccgtgctg gtgcacaaca atagagtgat cggagaggga
      961 tggaacaggc caatcggccg ccacgaccct accgcacacg cagagatcat ggcactgagg
     1021 cagggaggcc tggtcatgca gaattaccgc ctgatcgatg ccaccctgta tgtgacactg
     1081 gagccatgcg tgatgtgcgc aggagcaatg atccacagca ggatcggaag agtggtgttc
     1141 ggagcacggg acgccaagac cggcgcagca ggctccctga tggatgtgct gcaccacccc
     1201 ggcatgaacc accgggtgga gatcacagag ggaatcctgg cagacgagtg cgccgccctg
     1261 ctgagcgatt tctttagaat gcggagacag gagatcaagg cccagaagaa ggcacagagc
     1321 tccaccgact ctggaggatc tagcggagga tcctctggaa gcgagacacc aggcacaagc
     1381 gagtccgcca caccagagag ctccggcggc tcctccggag gatcctctga ggtggagttt
     1441 tcccacgagt actggatgag acatgccctg accctggcca agagggcacg cgatgagagg
     1501 gaggtgcctg tgggagccgt gctggtgctg aacaatagag tgatcggcga gggctggaac
     1561 agagccatcg gcctgcacga cccaacagcc catgccgaaa ttatggccct gagacagggc
     1621 ggcctggtca tgcagaacta cagactgatt gacgccaccc tgtacgtgac attcgagcct
     1681 tgcgtgatgt gcgccggcgc catgatccac tctaggatcg gccgcgtggt gtttggcgtg
     1741 aggaacgcaa aaaccggcgc cgcaggctcc ctgatggacg tgctgcacta ccccggcatg
     1801 aatcaccgcg tcgaaattac cgagggaatc ctggcagatg aatgtgccgc cctgctgtgc
     1861 tatttctttc ggatgcctag acaggtgttc aatgctcaga agaaggccca gagctccacc
     1921 gactccggag gatctagcgg aggctcctct ggctctgaga cacctggcac aagcgagagc
     1981 gcaacacctg aaagcagcgg gggcagcagc ggggggtcag ggaagcgaaa ttacattctg
     2041 gggctggcca ttggcattac atcagtgggc tatggcatca ttgactacga gacaagggac
     2101 gtgatcgacg ccggcgtgag actgttcaag gaggccaacg tggagaacaa tgagggccgg
     2161 agatccaaga ggggagcaag gcgcctgaag cggagaaggc gccacagaat ccagagagtg
     2221 aagaagctgc tgttcgatta caacctgctg accgaccact ccgagctgtc tggcatcaat
     2281 ccttatgagg ccagagtgaa gggcctgtcc cagaagctgt ctgaggagga gtttagcgcc
     2341 gccctgctgc acctggcaaa gaggagaggc gtgcacaacg tgaatgaggt ggaggaggac
     2401 accggcaacg agctgtccac aaaggagcag atcagccgca attccaaggc cctggaggag
     2461 aagtatgtgg ccgagctgca gctggagcgg ctgaagaagg atggcgaggt gaggggctcc
     2521 atcaatcgct tcaagacctc tgactacgtg aaggaggcca agcagctgct gaaggtgcag
     2581 aaggcctacc accagctgga tcagtccttt atcgatacat atatcgacct gctggagaca
     2641 aggcgcacat actatgaggg accaggagag ggctctccct tcggctggaa ggacatcaag
     2701 gagtggtacg agatgctgat gggccactgc acctattttc cagaggagct gagaagcgtg
     2761 aagtacgcct ataacgccga tctgtacaac gccctgaatg acctgaacaa cctggtcatc
     2821 accagggatg agaacgagaa gctggagtac tatgagaagt tccagatcat cgagaacgtg
     2881 ttcaagcaga agaagaagcc tacactgaag cagatcgcca aggagatcct ggtgaacgag
     2941 gaggacatca agggctaccg cgtgacctcc acaggcaagc cagagttcac caatctgaag
     3001 gtgtatcacg atatcaagga catcacagcc cggaaggaga tcatcgagaa cgccgagctg
     3061 ctggatcaga tcgccaagat cctgaccatc tatcagagct ccgaggacat ccaggaggag
     3121 ctgaccaacc tgaatagcga gctgacacag gaggagatcg agcagatcag caatctgaag
     3181 ggctacaccg gcacacacaa cctgagcctg aaggccatca atctgatcct ggatgagctg
     3241 tggcacacaa acgacaatca gatcgccatc tttaaccggc tgaagctggt gccaaagaag
     3301 gtggacctgt cccagcagaa ggagatccca accacactgg tggacgattt catcctgtct
     3361 cccgtggtga agcggagctt catccagagc atcaaagtga tcaacgccat catcaagaag
     3421 tacggcctgc ccaatgatat catcatcgag ctggccaggg agaagaactc caaggacgcc
     3481 cagaagatga tcaatgagat gcagaagagg aaccgccaga ccaatgagcg gatcgaggag
     3541 atcatcagaa ccacaggcaa ggagaacgcc aagtacctga tcgagaagat caagctgcac
     3601 gatatgcagg agggcaagtg tctgtattct ctggaggcca tccctctgga ggacctgctg
     3661 aacaatccat tcaactacga ggtggatcac atcatccccc ggagcgtgag cttcgacaat
     3721 tcttttaaca ataaggtgct ggtgaagcag gaggagaaca gcaagaaggg caataggacc
     3781 cctttccagt acctgtctag ctccgattct aagatcagct acgagacatt caagaagcac
     3841 atcctgaatc tggccaaggg caagggccgc atcagcaaga ccaagaagga gtacctgctg
     3901 gaggagcggg acatcaacag attctccgtg cagaaggact tcatcaaccg gaatctggtg
     3961 gacaccagat acgccacacg cggcctgatg aatctgctgc ggtcttattt cagagtgaac
     4021 aatctggatg tgaaggtgaa gagcatcaac ggcggcttca cctcctttct gcggagaaag
     4081 tggaagttta agaaggagcg caacaagggc tataagcacc acgccgagga tgccctgatc
     4141 atcgccaatg ccgacttcat ctttaaggag tggaagaagc tggacaaggc caagaaagtg
     4201 atggagaacc agatgttcga ggagaagcag gccgagagca tgcccgagat cgagacagag
     4261 caggagtaca aggagatttt catcacacct caccagatca agcacatcaa ggacttcaag
     4321 gactacaagt attctcacag ggtggataag aagcccaacc gcgagctgat caatgacacc
     4381 ctgtatagca cacggaagga cgataagggc aataccctga tcgtgaacaa tctgaacggc
     4441 ctgtacgaca aggataatga caagctgaag aagctgatca acaagtctcc cgagaagctg
     4501 ctgatgtacc accacgatcc tcagacatat cagaagctga agctgatcat ggagcagtac
     4561 ggcgacgaga agaacccact gtataagtac tatgaggaga caggcaacta cctgacaaag
     4621 tatagcaaga aggataatgg ccccgtgatc aagaagatca agtactatgg caacaagctg
     4681 aatgcccacc tggacatcac cgacgattac cctaactctc gcaataaggt ggtgaagctg
     4741 agcctgaagc cataccggtt cgacgtgtac ctggacaacg gcgtgtataa gtttgtgaca
     4801 gtgaagaatc tggatgtgat caagaaggag aactactatg aggtgaacag caagtgctac
     4861 gaggaggcca agaagctgaa gaagatcagc aaccaggccg agttcatcgc ctctttttac
     4921 aacaatgacc tgatcaagat caatggcgag ctgtatagag tgatcggcgt gaacaatgat
     4981 ctgctgaaca gaatcgaagt gaatatgatc gacatcacct acagggagta tctggagaac
     5041 atgaatgata agaggccccc tcgcatcatc aagaccatcg cctctaagac acagagcatc
     5101 aagaagtaca gcacagacat cctggggaac ctgtatgaag tcaagagcaa gaaacatcct
     5161 cagattatca agaaaggctc tggcggctca aaaagaaccg ccgacggcag cgaattcgag
     5221 cccaagaaga agaggaaagt ctaaccggtc atcatcacca tcaccattga gtttaaaccc
     5281 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     5341 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     5401 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     5461 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     5521 cttctgaggc ggaaagaacc agctggggct cgataccgtc gacctctagc tagagcttgg
     5581 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     5641 acatacgagc cggaagcata aagtgtaaag cctagggtgc ctaatgagtg agctaactca
     5701 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5761 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5821 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5881 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5941 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6001 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6061 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6121 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6181 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6241 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6301 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6361 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6421 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6481 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6541 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6601 ctacggggtc tgacactcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6661 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     6721 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6781 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6841 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6901 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6961 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7021 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7081 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7141 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7201 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7261 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7321 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7381 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7441 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7501 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7561 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     7621 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7681 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     7741 ctgacgtcga cggatcggga gatcgatctc ccgatcccct agggtcgact ctcagtacaa
     7801 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     7861 ctgagtagt
//
