LOCUS       sequence-436749-        8048 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SauriABE8e by MYL2 promoter and sgRNA targeting murine 
            Camk2d intron7 5' splice site by U6 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 8048)
  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 8048)
  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-U6-Camk2d sgRNA7
FEATURES             Location/Qualifiers
     source          1..8048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   22..151
                     /label=AAV2 ITR
     CDS             568..594
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             628..648
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4468..4488
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4502..4726
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(4858..5106)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(4916..4935)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(5086..5106)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     repeat_region   5130..5259
                     /label=AAV2 ITR
     primer_bind     complement(5278..5295)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5278..5294)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5287..5309)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5436..5891
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5523..5542)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5733..5754
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5917..6021
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6022..6090,6091..6882)
                     /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(6240..6259)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7053..7641
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7542..7561
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    7929..7950
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7965..7970,7971..7988,7989..7995)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8003..8019
                     /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     8008..8030
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8027..8043
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8027..8043
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttacgccaga tttaattaag gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag
       61 cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag
      121 ggagtggcca actccatcac taggggttcc tgcggcctct agaattcgct agctaggtct
      181 tgaaaggagt gggaattggc tccggtgccc gtcagtgggt tactcttccc cattagacaa
      241 tggcagggaa gagagcacac cccatcatcc ccaggccagg ccccagccac tgactcttta
      301 accttgaagg catttttggg tctcacgtgt ccacccaggc gggtggccgc ctttgagcag
      361 ctcttacttc agaagaacgg catggagtgg ggggtggggg gcttaggtgg cctccgcctc
      421 acctacaact gccaaaagtg gtcatggggt tatttttaac cccaggggag aggtatttat
      481 tgttccacag caggggcaga ggccagcagg ctcctcgaac tctccagagg tggcaactgg
      541 cctcagacac cgaccggtgc caccatgtac ccatacgatg ttccagatta cgctaaacgg
      601 acagccgacg gaagcgagtt cgagtcacca aagaagaagc ggaaagtctc tgaggtggag
      661 ttttcccacg agtactggat gagacatgcc ctgaccctgg ccaagagggc acgggatgag
      721 agggaggtgc ctgtgggagc cgtgctggtg ctgaacaata gagtgatcgg cgagggctgg
      781 aacagagcca tcggcctgca cgacccaaca gcccatgccg aaattatggc cctgagacag
      841 ggcggcctgg tcatgcagaa ctacagactg attgacgcca ccctgtacgt gacattcgag
      901 ccttgcgtga tgtgcgccgg cgccatgatc cactctagga tcggccgcgt ggtgtttggc
      961 gtgaggaact caaaaagagg cgccgcaggc tccctgatga acgtgctgaa ctaccccggc
     1021 atgaatcacc gcgtcgaaat taccgaggga atcctggcag atgaatgtgc cgccctgctg
     1081 tgcgatttct atcggatgcc tagacaggtg ttcaatgctc agaagaaggc ccagagctcc
     1141 atcaactccg gaggatctag cggaggctcc tctggctctg agacacctgg cacaagcgag
     1201 agcgcaacac ctgaaagcag cgggggcagc agcggggggt caatgcagga gaaccagcag
     1261 aagcaaaatt acatcctggg cctggccatc ggcatcacca gcgtgggcta tggcctgatc
     1321 gacagcaaga ccagagaagt gattgacgcc ggcgtgcggc tattcccaga ggccgactct
     1381 gaaaacaaca gcaatagaag atctaagcgg ggcgcccgga gactgaaaag acgaagaatc
     1441 cacagactga acagagtgaa agacctcctg gctgactacc agatgatcga cttaaacaac
     1501 gtgcccaagt ctaccgaccc ctacaccatc cgggtgaagg gactgcggga acctctgacc
     1561 aaggaagagt ttgccatcgc tctgctgcat atcgccaaga gaagaggcct gcacaacatc
     1621 tccgtgagca tgggcgatga ggaacaggac aacgagctgt ccaccaagca gcagctgcag
     1681 aagaacgctc agcagctgca ggacaaatac gtgtgcgagc tgcaactgga aagactgacc
     1741 aacatcaaca aggtcagagg cgagaagaac cggttcaaga ccgaagattt cgtgaaggaa
     1801 gtgaagcagc tgtgcgagac ccagcggcag taccacaaca tcgacgatca gttcatccag
     1861 cagtacatcg acctggtgag cacccggaga gaatacttcg agggccctgg caacggctct
     1921 ccatatggct gggatggaga tctgctgaag tggtatgaga agctgatggg cagatgcacc
     1981 tacttccctg aggagctgag aagcgtgaag tacgcctaca gcgccgatct gtttaacgcc
     2041 ctgaacgatc tgaacaacct ggttgtgacc cgcgacgaca accctaagct ggaatactac
     2101 gagaaatacc acattatcga gaacgtgttc aagcagaaga aaaatccaac tctgaagcaa
     2161 atcgccaaag agatcggcgt gcaggattac gacatcagag gatacagaat taccaagtcc
     2221 ggtaagcctc agttcaccag cttcaaactc taccacgacc tgaaaaacat ctttgaacag
     2281 gccaaatacc tggaagatgt ggagatgctg gatgagatag ctaaaatcct gacaatctac
     2341 caagacgaga tcagcatcaa gaaagccctg gaccagctgc ctgagctgct gaccgagagc
     2401 gaaaaaagcc agatcgctca gctgaccggc tacaccggta cacatagact gtctctgaag
     2461 tgcatccaca tcgtgatcga cgagctgtgg gagagccccg aaaaccagat ggaaatcttc
     2521 accagactga acctcaagcc aaagaaggtg gaaatgagcg agatcgacag catccctacc
     2581 acactggtgg atgagttcat cctgagccct gtggtgaagc gggccttcat ccagtccatc
     2641 aaggtgatca acgctgtgat caacagattc ggcctgcccg aggacatcat tatcgagctg
     2701 gccagagaga agaacagcaa ggacagaagg aagttcatca acaagctgca gaaacagaat
     2761 gaggccaccc ggaaaaaaat cgagcagctg ctggccaagt acggcaatac caatgccaag
     2821 tacatgatcg aaaagatcaa gctgcatgac atgcaggagg gcaagtgtct gtacagcctg
     2881 gaagctatcc ccctggaaga cctgctgtct aatcctacac actacgaggt ggaccacatc
     2941 atccctagaa gcgtgtcctt cgacaacagc ctgaacaaca aggttctggt gaagcaaagc
     3001 gagaacagca agaagggcaa taggacccct taccagtacc tgagcagcaa cgagtccaag
     3061 atctcttaca accagttcaa gcagcacatt ctgaacctgt ctaaggccaa agatagaatc
     3121 agcaagaaga aacgagatat gctgctggaa gaacgggaca tcaacaaatt cgaggtgcag
     3181 aaggaattca tcaacagaaa ccttgtggac acccggtacg ccactcggga gctgagcaac
     3241 ctgctgaaga cctacttcag cacacacgac tacgccgtga aagtgaagac catcaacggc
     3301 ggcttcacaa accacctgag gaaggtgtgg gacttcaaga agcaccggaa ccacggctac
     3361 aagcaccacg ccgaggatgc cctggtcatc gccaacgccg actttctgtt caaaacccac
     3421 aaggccctga gacggacaga caagatcctg gaacagcctg gactggaagt caacgacacc
     3481 accgtgaagg tggacacaga ggagaagtac caggagttat tcgagacacc gaaacaagtg
     3541 aagaacatca agcagtttag agatttcaag tattctcaca gagttgacaa gaagcccaac
     3601 cggcagctga tcaatgatac cctgtacagt accagagaga tcgatggcga aacctacgtg
     3661 gtccaaacac tgaaagacct gtacgccaag gacaatgaaa aggtgaaaaa gctctttaca
     3721 gaacggcctc aaaagatact gatgtaccag cacgatccta agacctttga gaaactgatg
     3781 accattctga atcagtacgc tgaggcaaag aatcctctgg ccgcttatta cgaggataag
     3841 ggcgaatacg tgaccaagta cgccaagaag ggcaacggcc ctgccatcca caagatcaaa
     3901 tacatcgaca agaaactggg cagctacctg gacgtgagta acaaatatcc tgagacacag
     3961 aacaagctgg tgaaactgtc tctgaagagc tttagattcg acatctacaa atgtgaacag
     4021 ggctacaaga tggtgtccat tggctacctc gacgtactga agaaggacaa ctactactac
     4081 atccccaaag ataagtacga ggccgagaag cagaaaaaga agatcaagga aagcgacctc
     4141 ttcgtgggca gcttctacta caacgacctg atcatgtacg aggacgaact cttccgggtg
     4201 atcggagtga actccgatat caacaacctg gttgagctga atatggtcga catcacctac
     4261 aaggatttct gcgaggtgaa caacgtgaca ggcgagaaga gaatcaagaa aaccatcgga
     4321 aagagagtgg tgctgatcga gaagtatacg accgacatcc tgggaaatct gtataaaacg
     4381 cccctgccta agaagcccca gctcattttc aagagaggcg agctgtctgg cggctcaaaa
     4441 agaaccgccg acggcagcga attcgagccc aagaagaaga ggaaagtcta atagatctcg
     4501 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     4561 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     4621 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     4681 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggctcg agcggcccaa
     4741 gcttaaaaaa atctcgccaa caagttgacg agataaacac ggcattttgc cttgttttag
     4801 tagattctgt aattttcatt acagagtact aaaaccaggc atggtttggt aagtagcggt
     4861 gtttcgtcct ttccacaaga tatataaagc caagaaatcg aaatactttc aagttacggt
     4921 aagcatatga tagtccattt taaaacataa ttttaaaact gcaaactacc caagaaatta
     4981 ttactttcta cgtcacgtat tttgtactaa tatctttgtg tttacagtca aattaattct
     5041 aattatctct ctaacagcct tgtatcgtat atgcaaatat gaaggaatca tgggaaatag
     5101 gccctcttcc tgcccgacct tgcggccgca ggaaccccta gtgatggagt tggccactcc
     5161 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     5221 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc cttaattaac ctaattcact
     5281 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     5341 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     5401 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     5461 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     5521 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5581 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5641 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     5701 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     5761 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     5821 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     5881 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5941 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     6001 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6061 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6121 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6181 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6241 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     6301 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6361 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6421 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6481 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6541 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6601 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6661 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6721 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6781 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6841 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6901 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6961 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     7021 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7081 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7141 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     7201 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7261 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7321 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7381 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7441 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7501 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7561 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7621 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7681 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     7741 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     7801 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     7861 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     7921 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     7981 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     8041 gaccatga
//
