LOCUS       sequence-411204-        7731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TadA7.10 and SpG cas9N by the constitutive CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7731)
  AUTHORS   Yang L, Liu Z, Sun J, Chen Z, Gao F, Guo Y
  TITLE     Adenine base editor-based correction of the cardiac pathogenic Lmna 
            c.1621C > T mutation in murine hearts.
  JOURNAL   J Cell Mol Med. 2024 Feb;28(4):e18145. doi: 10.1111/jcmm.18145.
  PUBMED    38332517
REFERENCE   2  (bases 1 to 7731)
  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: "J Cell Mol 
            Med."; date: "2024-02"; volume: "28(4):e18145. doi"; pages: " 
            10.1111/jcmm.18145"
COMMENT     Sequence Label: pAAV-CMV-TadA7.10-SpG N aa2-713-InteinN
FEATURES             Location/Qualifiers
     source          1..7731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   11..140
                     /label=AAV2 ITR
     primer_bind     complement(187..207)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     enhancer        255..634
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        635..838
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     789..809
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      866..875
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             878..898
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2087..3802
                     /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"
     primer_bind     complement(4542..4561)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    4571..4652
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     4596..4615
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4791..4920
                     /label=AAV2 ITR
     primer_bind     complement(4958..4974)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4958..4974)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4971..4993)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4982..4998
                     /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(5006..5012,5013..5030,5031..5036))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5051..5072
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5360..5948)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5440..5459)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6119..6910,6911..6979))
                     /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     6742..6761
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6980..7084)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7110..7565)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7247..7268)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7459..7478
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7692..7714
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7706..7723
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7707..7723
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ttaattaagg ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg
       61 ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa
      121 ctccatcact aggggttcct tgtagttaat gattaacccg ccatgctact tatctactgc
      181 ggccgcggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
      241 gtatgcaaag catggacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
      301 attagtccat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      361 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      421 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      481 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      541 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      601 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      661 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      721 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      781 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgt
      841 accgaagccg ctagcgctac cggtcgccac catggcccca aagaagaagc ggaaagtctc
      901 tgaagtcgag tttagccacg agtattggat gaggcacgca ctgaccctgg caaagcgagc
      961 atgggatgaa agagaagtcc ccgtgggcgc cgtgctggtg cacaacaata gagtgatcgg
     1021 agagggatgg aacaggccaa tcggccgcca cgaccctacc gcacacgcag agatcatggc
     1081 actgaggcag ggaggcctgg tcatgcagaa ttaccgcctg atcgatgcca ccctgtatgt
     1141 gacactggag ccatgcgtga tgtgcgcagg agcaatgatc cacagcagga tcggaagagt
     1201 ggtgttcgga gcacgggacg ccaagaccgg cgcagcaggc tccctgatgg atgtgctgca
     1261 ccaccccggc atgaaccacc gggtggagat cacagaggga atcctggcag acgagtgcgc
     1321 cgccctgctg agcgatttct ttagaatgcg gagacaggag atcaaggccc agaagaaggc
     1381 acagagctcc accgactctg gaggatctag cggaggatcc tctggaagcg agacaccagg
     1441 cacaagcgag tccgccacac cagagagctc cggcggctcc tccggaggat cctctgaggt
     1501 ggagttttcc cacgagtact ggatgagaca tgccctgacc ctggccaaga gggcacgcga
     1561 tgagagggag gtgcctgtgg gagccgtgct ggtgctgaac aatagagtga tcggcgaggg
     1621 ctggaacaga gccatcggcc tgcacgaccc aacagcccat gccgaaatta tggccctgag
     1681 acagggcggc ctggtcatgc agaactacag actgattgac gccaccctgt acgtgacatt
     1741 cgagccttgc gtgatgtgcg ccggcgccat gatccactct aggatcggcc gcgtggtgtt
     1801 tggcgtgagg aacgcaaaaa ccggcgccgc aggctccctg atggacgtgc tgcactaccc
     1861 cggcatgaat caccgcgtcg aaattaccga gggaatcctg gcagatgaat gtgccgccct
     1921 gctgtgctat ttctttcgga tgcctagaca ggtgttcaat gctcagaaga aggcccagag
     1981 ctccaccgac tccggaggat ctagcggagg ctcctctggc tctgagacac ctggcacaag
     2041 cgagagcgca acacctgaaa gcagcggggg cagcagcggg gggtcagaca agaagtacag
     2101 catcggcctg gccatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa
     2161 ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa
     2221 cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag
     2281 aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt
     2341 cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag agtccttcct
     2401 ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt
     2461 ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac
     2521 cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg
     2581 ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat
     2641 ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt
     2701 ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat
     2761 cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct
     2821 gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct
     2881 gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta
     2941 cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct
     3001 gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga
     3061 cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa
     3121 gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg
     3181 agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac
     3241 cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga
     3301 caacggcagc atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca
     3361 ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt
     3421 ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac
     3481 cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc
     3541 ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa
     3601 ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa
     3661 agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa
     3721 ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga
     3781 ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg
     3841 gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt
     3901 cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt
     3961 tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa
     4021 agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct
     4081 gatcaacggc atccgagaca agcagagtgg aaagacaatc ctggattttc ttaagtccga
     4141 tggatttgcc aaccggaact tcatgcagtt gatccatgat gactctctca cctttaagga
     4201 ggacatccag aaagcacaag tttgtctggc tggcgatact ctcattaccc tggccgatgg
     4261 acgacgagtg cctattagag aactggtgtc acagcagaat ttttccgtgt gggctctgaa
     4321 tcctcagact taccgcctgg agagggctag agtgagtaga gctttctgta ccggcatcaa
     4381 acctgtgtac cgcctcacca ctagactggg gagatccatt agggccactg ccaaccaccg
     4441 atttctcaca cctcagggct ggaaacgagt cgatgaactc cagcctggag attacctggc
     4501 tctgcctagg agaatcccta ctgcctcctg atctcgagct cgatgagttt ggacaaacca
     4561 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     4621 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     4681 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     4741 gtagcggccg caggtagata agtagcatgg cgggttaatc attaactaca aggaacccct
     4801 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     4861 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     4921 cctaattaag gccttaatta aatctggcgt aatcatggtc atagctgttt cctgtgtgaa
     4981 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5041 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5101 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     5161 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     5221 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5281 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5341 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5401 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5461 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5521 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5581 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5641 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5701 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5761 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5821 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5881 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5941 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6001 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6061 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6121 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6181 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     6241 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6301 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     6361 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6421 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6481 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6541 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6601 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6661 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6721 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6781 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6841 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6901 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6961 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7021 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7081 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     7141 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     7201 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     7261 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     7321 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     7381 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     7441 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     7501 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     7561 cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     7621 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     7681 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattag g
//
