LOCUS       sequence-369022-        8841 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ABEmax-SpCas9-NGX-loop.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8841)
  AUTHORS   Xu L, Zhang C, Li H, Wang P, Gao Y, Mokadam NA, Ma J, Arnold WD, Han
            R
  TITLE     Efficient precise in vivo base editing in adult dystrophic mice.
  JOURNAL   Nat Commun. 2021 Jun 17;12(1):3719. doi: 10.1038/s41467-021-23996-y.
  PUBMED    34140489
REFERENCE   2  (bases 1 to 8841)
  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 
            Commun."; date: "2021-06-17"; volume: "12(1):3719. doi"; pages: " 
            10.1038/s41467-021-23996-y"
COMMENT     Sequence Label: pCMV‐ABEmaxNGX‐loop
FEATURES             Location/Qualifiers
     source          1..8841
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(562..1353,1354..1422))
                     /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     1185..1204
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1423..1527)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1606..1625)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1797..2176
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2177..2380
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2331..2351
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2422..2441
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2422..2440
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2490..2510
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5331..7829
                     /codon_start=1
                     /product="C-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(C)
                     /translation="KPAFLSGDQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEI
                     SGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTY
                     AHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFIQLIH
                     DDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE
                     NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYL
                     QNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEV
                     VKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQI
                     LDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVV
                     GTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITL
                     ANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESIR
                     PKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIME
                     RSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARFLQKGNELALP
                     SKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLD
                     KVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPRAFKYFDTTIDRKVYRSTKEVLDATL
                     IHQSITGLYETRIDLSQLGGD"
     CDS             7872..7892
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7901..7918
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7941..7958)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7947..8171
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8242..8258)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8242..8258)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8255..8277)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8266..8282
                     /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(8290..8296,8297..8314,8315..8320))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8335..8356
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8473..8490)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(8644..8841,1..391))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8724..8743)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
       61 cgaacccccc gttcagcccg accgctgcgc cttatccagt aactatcgtc ttgagtccaa
      121 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
      181 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
      241 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
      301 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
      361 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
      421 tgacactcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
      481 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
      541 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
      601 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
      661 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
      721 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
      781 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
      841 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
      901 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
      961 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     1021 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     1081 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     1141 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     1201 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     1261 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     1321 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     1381 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     1441 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     1501 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     1561 cggatcggga gatcgatctc ccgatcccct agggtcgact ctcagtacaa tctgctctga
     1621 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     1681 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     1741 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
     1801 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     1861 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     1921 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     1981 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     2041 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     2101 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     2161 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     2221 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     2281 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     2341 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     2401 ccgctagaga tccgcggccg ctaatacgac tcactatagg gagagccgcc accatgaaac
     2461 ggacagccga cggaagcgag ttcgagtcac caaagaagaa gcggaaagtc tctgaagtcg
     2521 agtttagcca cgagtattgg atgaggcacg cactgaccct ggcaaagcga gcatgggatg
     2581 aaagagaagt ccccgtgggc gccgtgctgg tgcacaacaa tagagtgatc ggagagggat
     2641 ggaacaggcc aatcggccgc cacgacccta ccgcacacgc agagatcatg gcactgaggc
     2701 agggaggcct ggtcatgcag aattaccgcc tgatcgatgc caccctgtat gtgacactgg
     2761 agccatgcgt gatgtgcgca ggagcaatga tccacagcag gatcggaaga gtggtgttcg
     2821 gagcacggga cgccaagacc ggcgcagcag gctccctgat ggatgtgctg caccaccccg
     2881 gcatgaacca ccgggtggag atcacagagg gaatcctggc agacgagtgc gccgccctgc
     2941 tgagcgattt ctttagaatg cggagacagg agatcaaggc ccagaagaag gcacagagct
     3001 ccaccgactc tggaggatct agcggaggat cctctggaag cgagacacca ggcacaagcg
     3061 agtccgccac accagagagc tccggcggct cctccggagg atcctctgag gtggagtttt
     3121 cccacgagta ctggatgaga catgccctga ccctggccaa gagggcacgc gatgagaggg
     3181 aggtgcctgt gggagccgtg ctggtgctga acaatagagt gatcggcgag ggctggaaca
     3241 gagccatcgg cctgcacgac ccaacagccc atgccgaaat tatggccctg agacagggcg
     3301 gcctggtcat gcagaactac agactgattg acgccaccct gtacgtgaca ttcgagcctt
     3361 gcgtgatgtg cgccggcgcc atgatccact ctaggatcgg ccgcgtggtg tttggcgtga
     3421 ggaacgcaaa aaccggcgcc gcaggctccc tgatggacgt gctgcactac cccggcatga
     3481 atcaccgcgt cgaaattacc gagggaatcc tggcagatga atgtgccgcc ctgctgtgct
     3541 atttctttcg gatgcctaga caggtgttca atgctcagaa gaaggcccag agctccaccg
     3601 actccggagg atctagcgga ggctcctctg gctctgagac acctggcaca agcgagagcg
     3661 caacacctga aagcagcggg ggcagcagcg gggggtcaga caagaagtac agcatcggcc
     3721 tggccatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca
     3781 gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg
     3841 gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca
     3901 gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg
     3961 agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag
     4021 aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc
     4081 acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg
     4141 ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc
     4201 tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg
     4261 tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca
     4321 aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc
     4381 tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga
     4441 cccccaactt caagagcaac ttcgacctgg ccgaggatac caaactgcag ctgagcaagg
     4501 acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc
     4561 tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga
     4621 acaccgagat caccaaggcc cccctgagcg cctctatgat caagctgtac gacgagcacc
     4681 accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag
     4741 agattttctt cgaccagagc aagaacggct acgccggcta cgtgggcgcc gacaagaagc
     4801 tgcgcaagcg cagctctaaa ctggccacag aggaagagtt ctacaagttc atcaagccca
     4861 tcctggaaaa gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc
     4921 tgcggaagca gcggaccttc gacaacggca tcatccccca ccagatccac ctgggagagc
     4981 tgcacgccat tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa
     5041 agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa
     5101 acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg
     5161 agaaggtggt ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg
     5221 ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca
     5281 ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct
     5341 tcctgagcgg cgaccagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag
     5401 tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg
     5461 aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga
     5521 aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata
     5581 tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct
     5641 atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct
     5701 ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa
     5761 tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatccag ctgatccacg
     5821 acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata
     5881 gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc
     5941 agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca
     6001 tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg
     6061 agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac
     6121 accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg
     6181 ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg
     6241 accatatcgt gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca
     6301 gaagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga
     6361 tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca
     6421 atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga
     6481 gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga
     6541 tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga
     6601 agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca
     6661 acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca
     6721 aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc
     6781 ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct
     6841 acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga
     6901 agcggcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg
     6961 attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg
     7021 aggtgcagac aggcggcttc agcaaagagt ctatccggcc caagaggaac agcgataagc
     7081 tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgtc agccccaccg
     7141 tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga
     7201 gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca
     7261 tcgactttct ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc
     7321 ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgcccgct
     7381 ttctgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg
     7441 ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg
     7501 tggaacagca caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga
     7561 gagtgatcct ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg
     7621 ataagcccat cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg
     7681 gagcccctcg ggccttcaag tactttgaca ccaccatcga ccggaaggtg taccggagca
     7741 ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac
     7801 ggatcgacct gtctcagctg ggaggtgact ctggcggctc aaaaagaacc gccgacggca
     7861 gcgaattcga gcccaagaag aagaggaaag tctaaccggt catcatcacc atcaccattg
     7921 agtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     7981 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     8041 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     8101 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     8161 gggctctatg gcttctgagg cggaaagaac cagctggggc tcgataccgt cgacctctag
     8221 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     8281 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctagggtg cctaatgagt
     8341 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     8401 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     8461 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     8521 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     8581 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     8641 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     8701 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     8761 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     8821 aagcgtggcg ctttctcata g
//
