LOCUS       sequence-436789-        6112 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TadA8e and Sauri cas9N by MYL2 promoter, and add an HA tag
            fused to TadA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6112)
  AUTHORS   Liu Z, Yang L, Yang Y, Li J, Chen Z, Guo C, Guo Q, Li Q, Zhao D, Hu 
            X, Gao F, Guo Y
  TITLE     ABE-Mediated Cardiac Gene Silencing via Single AAVs Requires DNA 
            Accessibility.
  JOURNAL   Circ Res. 2025 Jan 16. doi: 10.1161/CIRCRESAHA.124.325611.
  PUBMED    39817340
REFERENCE   2  (bases 1 to 6112)
  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: "Circ Res. 
            2025 Jan 16. doi: 10.1161/CIRCRESAHA.124.325611."
COMMENT     Sequence Label: pAAV-MYL2-HA-TadA8e-Sauri N aa1-438-inteinN
FEATURES             Location/Qualifiers
     source          1..6112
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     143..162
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    530..551
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(566..571,572..589,590..596)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    604..620
                     /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)."
     primer_bind     609..631
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     628..644
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     628..644
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   682..811
                     /label=AAV2 ITR
     primer_bind     complement(858..878)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     regulatory      1307..1316
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1319..1345
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1346..1366
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3594..3613)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    3623..3704
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3648..3667
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3843..3972
                     /label=AAV2 ITR
     primer_bind     complement(3991..4008)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3991..4007)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4000..4022)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4149..4604
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4236..4255)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4446..4467
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4630..4734
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4735..4803,4804..5595)
                     /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     complement(4953..4972)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5766..6112,1..242)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
       61 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      121 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      181 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      241 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      301 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      361 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
      421 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
      481 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
      541 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
      601 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccagat
      661 ttaattaagg ccttaattag gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag
      721 cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag
      781 ggagtggcca actccatcac taggggttcc ttgtagttaa tgattaaccc gccatgctac
      841 ttatctactg cggccgcgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
      901 agcaggcaga agtatgcaaa gcatggacat tgattattga ctaggggtta ctcttcccca
      961 ttagacaatg gcagggaaga gagcacaccc catcatcccc aggccaggcc ccagccactg
     1021 actctttaac cttgaaggca tttttgggtc tcacgtgtcc acccaggcgg gtggccgcct
     1081 ttgagcagct cttacttcag aagaacggca tggagtgggg ggtggggggc ttaggtggcc
     1141 tccgcctcac ctacaactgc caaaagtggt catggggtta tttttaaccc caggggagag
     1201 gtatttattg ttccacagca ggggcagagg ccagcaggct cctcgaactc tccagaggtg
     1261 gcaactggcc tcagacaccg taccgaagcc gctagcgcta ccggtcgcca ccatggccta
     1321 cccatacgat gttccagatt acgctccaaa gaagaagcgg aaagtctctg aggtggagtt
     1381 ttcccacgag tactggatga gacatgccct gaccctggcc aagagggcac gggatgagag
     1441 ggaggtgcct gtgggagccg tgctggtgct gaacaataga gtgatcggcg agggctggaa
     1501 cagagccatc ggcctgcacg acccaacagc ccatgccgaa attatggccc tgagacaggg
     1561 cggcctggtc atgcagaact acagactgat tgacgccacc ctgtacgtga cattcgagcc
     1621 ttgcgtgatg tgcgccggcg ccatgatcca ctctaggatc ggccgcgtgg tgtttggcgt
     1681 gaggaactca aaaagaggcg ccgcaggctc cctgatgaac gtgctgaact accccggcat
     1741 gaatcaccgc gtcgaaatta ccgagggaat cctggcagat gaatgtgccg ccctgctgtg
     1801 cgatttctat cggatgccta gacaggtgtt caatgctcag aagaaggccc agagctccat
     1861 caactccgga ggatctagcg gaggctcctc tggctctgag acacctggca caagcgagag
     1921 cgcaacacct gaaagcagcg ggggcagcag cggggggtca atgcaggaga accagcagaa
     1981 gcaaaattac atcctgggcc tggccatcgg catcaccagc gtgggctatg gcctgatcga
     2041 cagcaagacc agagaagtga ttgacgccgg cgtgcggcta ttcccagagg ccgactctga
     2101 aaacaacagc aatagaagat ctaagcgggg cgcccggaga ctgaaaagac gaagaatcca
     2161 cagactgaac agagtgaaag acctcctggc tgactaccag atgatcgact taaacaacgt
     2221 gcccaagtct accgacccct acaccatccg ggtgaaggga ctgcgggaac ctctgaccaa
     2281 ggaagagttt gccatcgctc tgctgcatat cgccaagaga agaggcctgc acaacatctc
     2341 cgtgagcatg ggcgatgagg aacaggacaa cgagctgtcc accaagcagc agctgcagaa
     2401 gaacgctcag cagctgcagg acaaatacgt gtgcgagctg caactggaaa gactgaccaa
     2461 catcaacaag gtcagaggcg agaagaaccg gttcaagacc gaagatttcg tgaaggaagt
     2521 gaagcagctg tgcgagaccc agcggcagta ccacaacatc gacgatcagt tcatccagca
     2581 gtacatcgac ctggtgagca cccggagaga atacttcgag ggccctggca acggctctcc
     2641 atatggctgg gatggagatc tgctgaagtg gtatgagaag ctgatgggca gatgcaccta
     2701 cttccctgag gagctgagaa gcgtgaagta cgcctacagc gccgatctgt ttaacgccct
     2761 gaacgatctg aacaacctgg ttgtgacccg cgacgacaac cctaagctgg aatactacga
     2821 gaaataccac attatcgaga acgtgttcaa gcagaagaaa aatccaactc tgaagcaaat
     2881 cgccaaagag atcggcgtgc aggattacga catcagagga tacagaatta ccaagtccgg
     2941 taagcctcag ttcaccagct tcaaactcta ccacgacctg aaaaacatct ttgaacaggc
     3001 caaatacctg gaagatgtgg agatgctgga tgagatagct aaaatcctga caatctacca
     3061 agacgagatc agcatcaaga aagccctgga ccagctgcct gagctgctga ccgagagcga
     3121 aaaaagccag atcgctcagc tgaccggcta caccggtaca catagactgt ctctgaagtg
     3181 catccacatc gtgatcgacg agctgtggga gagccccgaa aaccagatgg aaatcttcac
     3241 cagactgaac ctcaagccaa agaaggtgga aatgtgtctg gctggcgata ctctcattac
     3301 cctggccgat ggacgacgag tgcctattag agaactggtg tcacagcaga atttttccgt
     3361 gtgggctctg aatcctcaga cttaccgcct ggagagggct agagtgagta gagctttctg
     3421 taccggcatc aaacctgtgt accgcctcac cactagactg gggagatcca ttagggccac
     3481 tgccaaccac cgatttctca cacctcaggg ctggaaacga gtcgatgaac tccagcctgg
     3541 agattacctg gctctgccta ggagaatccc tactgcctcc tgatctcgag ctcgatgagt
     3601 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     3661 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     3721 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     3781 tctacaaatg tggtagcggc cgcaggtaga taagtagcat ggcgggttaa tcattaacta
     3841 caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     3901 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
     3961 gcgagcgcgc agccttaatt aacctaattc actggccgtc gttttacaac gtcgtgactg
     4021 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     4081 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     4141 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4201 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4261 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4321 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4381 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4441 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4501 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4561 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     4621 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4681 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     4741 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     4801 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     4861 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     4921 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     4981 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5041 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5101 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5161 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5221 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5281 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5341 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5401 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5461 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5521 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5581 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5641 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     5701 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     5761 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     5821 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     5881 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     5941 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6001 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6061 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag ct
//
