LOCUS       sequence-231806-        8871 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 8871)
  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 8871)
  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: CP1012-ABEmax
FEATURES             Location/Qualifiers
     source          1..8871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        113..492
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        493..696
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     647..667
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     738..757
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        738..756
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             806..826
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3146..4861
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
     CDS             6218..6238
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6247..6264
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6287..6304)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6293..6517
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6588..6604)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6588..6604)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6601..6623)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6612..6628
                     /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(6636..6642,6643..6660,6661..6666))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6681..6702
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6819..6836)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6990..7578)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7070..7089)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7749..8540,8541..8609))
                     /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     8372..8391
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8610..8714)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8793..8812)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
       61 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
      121 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
      181 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      241 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      301 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      361 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      421 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      481 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      541 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      601 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      661 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
      721 tagagatccg cggccgctaa tacgactcac tatagggaga gccgccacca tgaaacggac
      781 agccgacgga agcgagttcg agtcaccaaa gaagaagcgg aaagtctctg aagtcgagtt
      841 tagccacgag tattggatga ggcacgcact gaccctggca aagcgagcat gggatgaaag
      901 agaagtcccc gtgggcgccg tgctggtgca caacaataga gtgatcggag agggatggaa
      961 caggccaatc ggccgccacg accctaccgc acacgcagag atcatggcac tgaggcaggg
     1021 aggcctggtc atgcagaatt accgcctgat cgatgccacc ctgtatgtga cactggagcc
     1081 atgcgtgatg tgcgcaggag caatgatcca cagcaggatc ggaagagtgg tgttcggagc
     1141 acgggacgcc aagaccggcg cagcaggctc cctgatggat gtgctgcacc accccggcat
     1201 gaaccaccgg gtggagatca cagagggaat cctggcagac gagtgcgccg ccctgctgag
     1261 cgatttcttt agaatgcgga gacaggagat caaggcccag aagaaggcac agagctccac
     1321 cgactctgga ggatctagcg gaggatcctc tggaagcgag acaccaggca caagcgagtc
     1381 cgccacacca gagagctccg gcggctcctc cggaggatcc tctgaggtgg agttttccca
     1441 cgagtactgg atgagacatg ccctgaccct ggccaagagg gcacgcgatg agagggaggt
     1501 gcctgtggga gccgtgctgg tgctgaacaa tagagtgatc ggcgagggct ggaacagagc
     1561 catcggcctg cacgacccaa cagcccatgc cgaaattatg gccctgagac agggcggcct
     1621 ggtcatgcag aactacagac tgattgacgc caccctgtac gtgacattcg agccttgcgt
     1681 gatgtgcgcc ggcgccatga tccactctag gatcggccgc gtggtgtttg gcgtgaggaa
     1741 cgcaaaaacc ggcgccgcag gctccctgat ggacgtgctg cactaccccg gcatgaatca
     1801 ccgcgtcgaa attaccgagg gaatcctggc agatgaatgt gccgccctgc tgtgctattt
     1861 ctttcggatg cctagacagg tgttcaatgc tcagaagaag gcccagagct ccaccgactc
     1921 cggaggatct agcggaggct cctctggctc tgagacacct ggcacaagcg agagcgcaac
     1981 acctgaaagc agcgggggca gcagcggggg gtcagactac aaggtgtacg acgtgcggaa
     2041 gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag
     2101 caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg
     2161 gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt
     2221 tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt
     2281 gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat
     2341 cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc
     2401 ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt
     2461 gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga
     2521 ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa
     2581 gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact
     2641 gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag
     2701 ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga
     2761 acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt
     2821 gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa
     2881 gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc
     2941 ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa
     3001 agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat
     3061 cgacctgtct cagctgggag gcgacggcgg atcaggaggc tctggcggtt caggtggatc
     3121 aggcggtagc ggaggttcag gtggtgacaa gaagtacagc atcggcctgg ccatcggcac
     3181 caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa
     3241 ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt
     3301 cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac
     3361 cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt
     3421 ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca
     3481 cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc
     3541 caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct
     3601 gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga
     3661 cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa
     3721 ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc
     3781 tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa
     3841 gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa
     3901 gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga
     3961 cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc
     4021 caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac
     4081 caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac
     4141 cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga
     4201 ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta
     4261 caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct
     4321 gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca
     4381 gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct
     4441 gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg
     4501 ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat
     4561 caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga
     4621 gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct
     4681 gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg
     4741 aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt
     4801 caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga
     4861 gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac
     4921 ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga
     4981 ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga
     5041 ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg
     5101 gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa
     5161 gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt
     5221 catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt
     5281 gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat
     5341 taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg
     5401 gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg
     5461 acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag
     5521 ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct
     5581 gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct
     5641 gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg actccatcga
     5701 caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga
     5761 agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac
     5821 ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa
     5881 ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca
     5941 gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt
     6001 gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta
     6061 caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt
     6121 gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggctctgg
     6181 cggctcaaaa agaaccgccg acggcagcga attcgagccc aagaagaaga ggaaagtcta
     6241 accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct
     6301 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     6361 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     6421 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     6481 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     6541 tggggctcga taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     6601 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     6661 tgtaaagcct agggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     6721 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6781 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6841 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6901 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6961 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     7021 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     7081 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     7141 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     7201 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     7261 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     7321 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     7381 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     7441 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     7501 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7561 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cactcagtgg
     7621 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7681 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7741 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     7801 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     7861 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7921 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7981 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     8041 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     8101 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     8161 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     8221 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     8281 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     8341 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     8401 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     8461 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     8521 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     8581 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     8641 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     8701 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     8761 cgatctcccg atcccctagg gtcgactctc agtacaatct gctctgatgc cgcatagtta
     8821 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc g
//
