LOCUS       sequence-436679-        8006 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SauriABE8e by EFS promoter and sgRNA targeting murine 
            Camk2d start codon site by U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8006)
  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 8006)
  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-EFS-TadA8e-Sauri-U6-Camk2d sgRNA1
FEATURES             Location/Qualifiers
     source          1..8006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(136..153)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(136..152)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(145..167)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      294..749
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(381..400)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     591..612
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        775..879
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(880..948,949..1740)
                     /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(1098..1117)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1911..2499
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2400..2419
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    2787..2808
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2823..2828,2829..2846,2847..2853)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2861..2877
                     /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     2866..2888
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2885..2901
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2885..2901
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   2928..3057
                     /label=AAV2 ITR
     promoter        3123..3334
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             3384..3404
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7224..7244
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    7258..7482
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(7614..7862)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(7672..7691)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(7842..7862)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
       61 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagcct
      121 taattaacct aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
      181 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
      241 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
      301 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
      361 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
      421 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
      481 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
      541 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
      601 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
      661 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
      721 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
      781 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
      841 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
      901 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
      961 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     1021 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1081 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     1141 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     1201 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     1261 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     1321 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     1381 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     1441 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     1501 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     1561 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     1621 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     1681 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     1741 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     1801 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     1861 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1921 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1981 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2041 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2101 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2161 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2221 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2281 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2341 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2401 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2461 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     2521 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     2581 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     2641 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     2701 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     2761 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     2821 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     2881 cacacaggaa acagctatga ccatgattac gccagattta attaaggctg cgcgctcgct
     2941 cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct
     3001 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg
     3061 gcctctagaa ttcgctagct aggtcttgaa aggagtggga attggctccg gtgcccgtca
     3121 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
     3181 atccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
     3241 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
     3301 tctttttcgc aacgggtttg ccgccagaac acaggaccgg tgccaccatg aaacggacag
     3361 ccgacggaag cgagttcgag tcaccaaaga agaagcggaa agtctctgag gtggagtttt
     3421 cccacgagta ctggatgaga catgccctga ccctggccaa gagggcacgg gatgagaggg
     3481 aggtgcctgt gggagccgtg ctggtgctga acaatagagt gatcggcgag ggctggaaca
     3541 gagccatcgg cctgcacgac ccaacagccc atgccgaaat tatggccctg agacagggcg
     3601 gcctggtcat gcagaactac agactgattg acgccaccct gtacgtgaca ttcgagcctt
     3661 gcgtgatgtg cgccggcgcc atgatccact ctaggatcgg ccgcgtggtg tttggcgtga
     3721 ggaactcaaa aagaggcgcc gcaggctccc tgatgaacgt gctgaactac cccggcatga
     3781 atcaccgcgt cgaaattacc gagggaatcc tggcagatga atgtgccgcc ctgctgtgcg
     3841 atttctatcg gatgcctaga caggtgttca atgctcagaa gaaggcccag agctccatca
     3901 actccggagg atctagcgga ggctcctctg gctctgagac acctggcaca agcgagagcg
     3961 caacacctga aagcagcggg ggcagcagcg gggggtcaat gcaggagaac cagcagaagc
     4021 aaaattacat cctgggcctg gccatcggca tcaccagcgt gggctatggc ctgatcgaca
     4081 gcaagaccag agaagtgatt gacgccggcg tgcggctatt cccagaggcc gactctgaaa
     4141 acaacagcaa tagaagatct aagcggggcg cccggagact gaaaagacga agaatccaca
     4201 gactgaacag agtgaaagac ctcctggctg actaccagat gatcgactta aacaacgtgc
     4261 ccaagtctac cgacccctac accatccggg tgaagggact gcgggaacct ctgaccaagg
     4321 aagagtttgc catcgctctg ctgcatatcg ccaagagaag aggcctgcac aacatctccg
     4381 tgagcatggg cgatgaggaa caggacaacg agctgtccac caagcagcag ctgcagaaga
     4441 acgctcagca gctgcaggac aaatacgtgt gcgagctgca actggaaaga ctgaccaaca
     4501 tcaacaaggt cagaggcgag aagaaccggt tcaagaccga agatttcgtg aaggaagtga
     4561 agcagctgtg cgagacccag cggcagtacc acaacatcga cgatcagttc atccagcagt
     4621 acatcgacct ggtgagcacc cggagagaat acttcgaggg ccctggcaac ggctctccat
     4681 atggctggga tggagatctg ctgaagtggt atgagaagct gatgggcaga tgcacctact
     4741 tccctgagga gctgagaagc gtgaagtacg cctacagcgc cgatctgttt aacgccctga
     4801 acgatctgaa caacctggtt gtgacccgcg acgacaaccc taagctggaa tactacgaga
     4861 aataccacat tatcgagaac gtgttcaagc agaagaaaaa tccaactctg aagcaaatcg
     4921 ccaaagagat cggcgtgcag gattacgaca tcagaggata cagaattacc aagtccggta
     4981 agcctcagtt caccagcttc aaactctacc acgacctgaa aaacatcttt gaacaggcca
     5041 aatacctgga agatgtggag atgctggatg agatagctaa aatcctgaca atctaccaag
     5101 acgagatcag catcaagaaa gccctggacc agctgcctga gctgctgacc gagagcgaaa
     5161 aaagccagat cgctcagctg accggctaca ccggtacaca tagactgtct ctgaagtgca
     5221 tccacatcgt gatcgacgag ctgtgggaga gccccgaaaa ccagatggaa atcttcacca
     5281 gactgaacct caagccaaag aaggtggaaa tgagcgagat cgacagcatc cctaccacac
     5341 tggtggatga gttcatcctg agccctgtgg tgaagcgggc cttcatccag tccatcaagg
     5401 tgatcaacgc tgtgatcaac agattcggcc tgcccgagga catcattatc gagctggcca
     5461 gagagaagaa cagcaaggac agaaggaagt tcatcaacaa gctgcagaaa cagaatgagg
     5521 ccacccggaa aaaaatcgag cagctgctgg ccaagtacgg caataccaat gccaagtaca
     5581 tgatcgaaaa gatcaagctg catgacatgc aggagggcaa gtgtctgtac agcctggaag
     5641 ctatccccct ggaagacctg ctgtctaatc ctacacacta cgaggtggac cacatcatcc
     5701 ctagaagcgt gtccttcgac aacagcctga acaacaaggt tctggtgaag caaagcgaga
     5761 acagcaagaa gggcaatagg accccttacc agtacctgag cagcaacgag tccaagatct
     5821 cttacaacca gttcaagcag cacattctga acctgtctaa ggccaaagat agaatcagca
     5881 agaagaaacg agatatgctg ctggaagaac gggacatcaa caaattcgag gtgcagaagg
     5941 aattcatcaa cagaaacctt gtggacaccc ggtacgccac tcgggagctg agcaacctgc
     6001 tgaagaccta cttcagcaca cacgactacg ccgtgaaagt gaagaccatc aacggcggct
     6061 tcacaaacca cctgaggaag gtgtgggact tcaagaagca ccggaaccac ggctacaagc
     6121 accacgccga ggatgccctg gtcatcgcca acgccgactt tctgttcaaa acccacaagg
     6181 ccctgagacg gacagacaag atcctggaac agcctggact ggaagtcaac gacaccaccg
     6241 tgaaggtgga cacagaggag aagtaccagg agttattcga gacaccgaaa caagtgaaga
     6301 acatcaagca gtttagagat ttcaagtatt ctcacagagt tgacaagaag cccaaccggc
     6361 agctgatcaa tgataccctg tacagtacca gagagatcga tggcgaaacc tacgtggtcc
     6421 aaacactgaa agacctgtac gccaaggaca atgaaaaggt gaaaaagctc tttacagaac
     6481 ggcctcaaaa gatactgatg taccagcacg atcctaagac ctttgagaaa ctgatgacca
     6541 ttctgaatca gtacgctgag gcaaagaatc ctctggccgc ttattacgag gataagggcg
     6601 aatacgtgac caagtacgcc aagaagggca acggccctgc catccacaag atcaaataca
     6661 tcgacaagaa actgggcagc tacctggacg tgagtaacaa atatcctgag acacagaaca
     6721 agctggtgaa actgtctctg aagagcttta gattcgacat ctacaaatgt gaacagggct
     6781 acaagatggt gtccattggc tacctcgacg tactgaagaa ggacaactac tactacatcc
     6841 ccaaagataa gtacgaggcc gagaagcaga aaaagaagat caaggaaagc gacctcttcg
     6901 tgggcagctt ctactacaac gacctgatca tgtacgagga cgaactcttc cgggtgatcg
     6961 gagtgaactc cgatatcaac aacctggttg agctgaatat ggtcgacatc acctacaagg
     7021 atttctgcga ggtgaacaac gtgacaggcg agaagagaat caagaaaacc atcggaaaga
     7081 gagtggtgct gatcgagaag tatacgaccg acatcctggg aaatctgtat aaaacgcccc
     7141 tgcctaagaa gccccagctc attttcaaga gaggcgagct gtctggcggc tcaaaaagaa
     7201 ccgccgacgg cagcgaattc gagcccaaga agaagaggaa agtctaatag atctcgactg
     7261 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     7321 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     7381 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     7441 aagacaatag caggcatgct ggggatgcgg tgggctctat ggctcgagcg gcccaagctt
     7501 aaaaaaatct cgccaacaag ttgacgagat aaacacggca ttttgccttg ttttagtaga
     7561 ttctgtaatt ttcattacag agtactaaaa caccccgcca tggcttcgac cacggtgttt
     7621 cgtcctttcc acaagatata taaagccaag aaatcgaaat actttcaagt tacggtaagc
     7681 atatgatagt ccattttaaa acataatttt aaaactgcaa actacccaag aaattattac
     7741 tttctacgtc acgtattttg tactaatatc tttgtgttta cagtcaaatt aattctaatt
     7801 atctctctaa cagccttgta tcgtatatgc aaatatgaag gaatcatggg aaataggccc
     7861 tcttcctgcc cgaccttgcg gccgcaggaa cccctagtga tggagttggc cactccctct
     7921 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     7981 gcccgggcgg cctcagtgag cgagcg
//
