LOCUS       sequence-436705-        7999 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.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7999)
  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 7999)
  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-TadA8e-Sauri-U6-Camk2d sgRNA7
FEATURES             Location/Qualifiers
     source          1..7999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             579..599
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4419..4439
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4453..4677
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(4809..5057)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(4867..4886)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(5037..5057)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     repeat_region   5081..5210
                     /label=AAV2 ITR
     primer_bind     complement(5229..5246)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5229..5245)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5238..5260)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5387..5842
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5474..5493)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5684..5705
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5868..5972
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5973..6041,6042..6833)
                     /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(6191..6210)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7004..7592
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7493..7512
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    7880..7901
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7916..7921,7922..7939,7940..7946)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7954..7970
                     /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     7959..7981
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7978..7994
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7978..7994
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttacgccaga tttaattaag gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag
       61 cccgggcggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta
      121 ggggttcctg cggcctctag aattcgctag ctaggtcttg aaaggagtgg gaattggctc
      181 cggtgcccgt cagtgggtta ctcttcccca ttagacaatg gcagggaaga gagcacaccc
      241 catcatcccc aggccaggcc ccagccactg actctttaac cttgaaggca tttttgggtc
      301 tcacgtgtcc acccaggcgg gtggccgcct ttgagcagct cttacttcag aagaacggca
      361 tggagtgggg ggtggggggc ttaggtggcc tccgcctcac ctacaactgc caaaagtggt
      421 catggggtta tttttaaccc caggggagag gtatttattg ttccacagca ggggcagagg
      481 ccagcaggct cctcgaactc tccagaggtg gcaactggcc tcagacaccg accggtgcca
      541 ccatgaaacg gacagccgac ggaagcgagt tcgagtcacc aaagaagaag cggaaagtct
      601 ctgaggtgga gttttcccac gagtactgga tgagacatgc cctgaccctg gccaagaggg
      661 cacgggatga gagggaggtg cctgtgggag ccgtgctggt gctgaacaat agagtgatcg
      721 gcgagggctg gaacagagcc atcggcctgc acgacccaac agcccatgcc gaaattatgg
      781 ccctgagaca gggcggcctg gtcatgcaga actacagact gattgacgcc accctgtacg
      841 tgacattcga gccttgcgtg atgtgcgccg gcgccatgat ccactctagg atcggccgcg
      901 tggtgtttgg cgtgaggaac tcaaaaagag gcgccgcagg ctccctgatg aacgtgctga
      961 actaccccgg catgaatcac cgcgtcgaaa ttaccgaggg aatcctggca gatgaatgtg
     1021 ccgccctgct gtgcgatttc tatcggatgc ctagacaggt gttcaatgct cagaagaagg
     1081 cccagagctc catcaactcc ggaggatcta gcggaggctc ctctggctct gagacacctg
     1141 gcacaagcga gagcgcaaca cctgaaagca gcgggggcag cagcgggggg tcaatgcagg
     1201 agaaccagca gaagcaaaat tacatcctgg gcctggccat cggcatcacc agcgtgggct
     1261 atggcctgat cgacagcaag accagagaag tgattgacgc cggcgtgcgg ctattcccag
     1321 aggccgactc tgaaaacaac agcaatagaa gatctaagcg gggcgcccgg agactgaaaa
     1381 gacgaagaat ccacagactg aacagagtga aagacctcct ggctgactac cagatgatcg
     1441 acttaaacaa cgtgcccaag tctaccgacc cctacaccat ccgggtgaag ggactgcggg
     1501 aacctctgac caaggaagag tttgccatcg ctctgctgca tatcgccaag agaagaggcc
     1561 tgcacaacat ctccgtgagc atgggcgatg aggaacagga caacgagctg tccaccaagc
     1621 agcagctgca gaagaacgct cagcagctgc aggacaaata cgtgtgcgag ctgcaactgg
     1681 aaagactgac caacatcaac aaggtcagag gcgagaagaa ccggttcaag accgaagatt
     1741 tcgtgaagga agtgaagcag ctgtgcgaga cccagcggca gtaccacaac atcgacgatc
     1801 agttcatcca gcagtacatc gacctggtga gcacccggag agaatacttc gagggccctg
     1861 gcaacggctc tccatatggc tgggatggag atctgctgaa gtggtatgag aagctgatgg
     1921 gcagatgcac ctacttccct gaggagctga gaagcgtgaa gtacgcctac agcgccgatc
     1981 tgtttaacgc cctgaacgat ctgaacaacc tggttgtgac ccgcgacgac aaccctaagc
     2041 tggaatacta cgagaaatac cacattatcg agaacgtgtt caagcagaag aaaaatccaa
     2101 ctctgaagca aatcgccaaa gagatcggcg tgcaggatta cgacatcaga ggatacagaa
     2161 ttaccaagtc cggtaagcct cagttcacca gcttcaaact ctaccacgac ctgaaaaaca
     2221 tctttgaaca ggccaaatac ctggaagatg tggagatgct ggatgagata gctaaaatcc
     2281 tgacaatcta ccaagacgag atcagcatca agaaagccct ggaccagctg cctgagctgc
     2341 tgaccgagag cgaaaaaagc cagatcgctc agctgaccgg ctacaccggt acacatagac
     2401 tgtctctgaa gtgcatccac atcgtgatcg acgagctgtg ggagagcccc gaaaaccaga
     2461 tggaaatctt caccagactg aacctcaagc caaagaaggt ggaaatgagc gagatcgaca
     2521 gcatccctac cacactggtg gatgagttca tcctgagccc tgtggtgaag cgggccttca
     2581 tccagtccat caaggtgatc aacgctgtga tcaacagatt cggcctgccc gaggacatca
     2641 ttatcgagct ggccagagag aagaacagca aggacagaag gaagttcatc aacaagctgc
     2701 agaaacagaa tgaggccacc cggaaaaaaa tcgagcagct gctggccaag tacggcaata
     2761 ccaatgccaa gtacatgatc gaaaagatca agctgcatga catgcaggag ggcaagtgtc
     2821 tgtacagcct ggaagctatc cccctggaag acctgctgtc taatcctaca cactacgagg
     2881 tggaccacat catccctaga agcgtgtcct tcgacaacag cctgaacaac aaggttctgg
     2941 tgaagcaaag cgagaacagc aagaagggca ataggacccc ttaccagtac ctgagcagca
     3001 acgagtccaa gatctcttac aaccagttca agcagcacat tctgaacctg tctaaggcca
     3061 aagatagaat cagcaagaag aaacgagata tgctgctgga agaacgggac atcaacaaat
     3121 tcgaggtgca gaaggaattc atcaacagaa accttgtgga cacccggtac gccactcggg
     3181 agctgagcaa cctgctgaag acctacttca gcacacacga ctacgccgtg aaagtgaaga
     3241 ccatcaacgg cggcttcaca aaccacctga ggaaggtgtg ggacttcaag aagcaccgga
     3301 accacggcta caagcaccac gccgaggatg ccctggtcat cgccaacgcc gactttctgt
     3361 tcaaaaccca caaggccctg agacggacag acaagatcct ggaacagcct ggactggaag
     3421 tcaacgacac caccgtgaag gtggacacag aggagaagta ccaggagtta ttcgagacac
     3481 cgaaacaagt gaagaacatc aagcagttta gagatttcaa gtattctcac agagttgaca
     3541 agaagcccaa ccggcagctg atcaatgata ccctgtacag taccagagag atcgatggcg
     3601 aaacctacgt ggtccaaaca ctgaaagacc tgtacgccaa ggacaatgaa aaggtgaaaa
     3661 agctctttac agaacggcct caaaagatac tgatgtacca gcacgatcct aagacctttg
     3721 agaaactgat gaccattctg aatcagtacg ctgaggcaaa gaatcctctg gccgcttatt
     3781 acgaggataa gggcgaatac gtgaccaagt acgccaagaa gggcaacggc cctgccatcc
     3841 acaagatcaa atacatcgac aagaaactgg gcagctacct ggacgtgagt aacaaatatc
     3901 ctgagacaca gaacaagctg gtgaaactgt ctctgaagag ctttagattc gacatctaca
     3961 aatgtgaaca gggctacaag atggtgtcca ttggctacct cgacgtactg aagaaggaca
     4021 actactacta catccccaaa gataagtacg aggccgagaa gcagaaaaag aagatcaagg
     4081 aaagcgacct cttcgtgggc agcttctact acaacgacct gatcatgtac gaggacgaac
     4141 tcttccgggt gatcggagtg aactccgata tcaacaacct ggttgagctg aatatggtcg
     4201 acatcaccta caaggatttc tgcgaggtga acaacgtgac aggcgagaag agaatcaaga
     4261 aaaccatcgg aaagagagtg gtgctgatcg agaagtatac gaccgacatc ctgggaaatc
     4321 tgtataaaac gcccctgcct aagaagcccc agctcatttt caagagaggc gagctgtctg
     4381 gcggctcaaa aagaaccgcc gacggcagcg aattcgagcc caagaagaag aggaaagtct
     4441 aatagatctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     4501 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     4561 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     4621 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctc
     4681 gagcggccca agcttaaaaa aatctcgcca acaagttgac gagataaaca cggcattttg
     4741 ccttgtttta gtagattctg taattttcat tacagagtac taaaaccagg catggtttgg
     4801 taagtagcgg tgtttcgtcc tttccacaag atatataaag ccaagaaatc gaaatacttt
     4861 caagttacgg taagcatatg atagtccatt ttaaaacata attttaaaac tgcaaactac
     4921 ccaagaaatt attactttct acgtcacgta ttttgtacta atatctttgt gtttacagtc
     4981 aaattaattc taattatctc tctaacagcc ttgtatcgta tatgcaaata tgaaggaatc
     5041 atgggaaata ggccctcttc ctgcccgacc ttgcggccgc aggaacccct agtgatggag
     5101 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     5161 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ccttaattaa
     5221 cctaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     5281 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     5341 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     5401 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     5461 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     5521 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     5581 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     5641 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     5701 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     5761 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     5821 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5881 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5941 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     6001 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     6061 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     6121 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     6181 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     6241 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     6301 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     6361 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     6421 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     6481 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     6541 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     6601 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     6661 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6721 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6781 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6841 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6901 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6961 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     7021 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     7081 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     7141 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     7201 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     7261 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     7321 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     7381 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     7441 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     7501 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     7561 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     7621 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     7681 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     7741 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     7801 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     7861 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     7921 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     7981 gaaacagcta tgaccatga
//
