LOCUS       sequence-436692-        7898 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses SauriABE8e by EFS 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 7898)
  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 7898)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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 sgRNA7
FEATURES             Location/Qualifiers
     source          1..7898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     primer_bind     complement(149..166)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(149..165)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(158..180)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      307..762
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(394..413)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     604..625
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        788..892
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(893..961,962..1753)
                     /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(1111..1130)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1924..2512
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2413..2432
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    2800..2821
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2836..2841,2842..2859,2860..2866)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2874..2890
                     /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     2879..2901
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2898..2914
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2898..2914
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   2941..3070
                     /label=AAV2 ITR
     promoter        3136..3347
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             3397..3417
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7237..7257
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    7271..7495
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(7627..7875)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(7685..7704)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(7855..7875)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
       61 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
      121 gagcgcgcag ccttaattaa cctaattcac tggccgtcgt tttacaacgt cgtgactggg
      181 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
      241 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
      301 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
      361 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
      421 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
      481 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
      541 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
      601 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
      661 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
      721 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
      781 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
      841 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
      901 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
      961 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1021 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     1081 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     1141 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1201 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     1261 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     1321 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     1381 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     1441 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     1501 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     1561 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     1621 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     1681 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     1741 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     1801 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     1861 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     1921 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     1981 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2041 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     2101 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2161 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2221 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2281 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2341 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2401 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2461 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2521 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     2581 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     2641 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     2701 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     2761 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     2821 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     2881 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccagat ttaattaagg
     2941 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
     3001 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
     3061 aggggttcct gcggcctcta gaattcgcta gctaggtctt gaaaggagtg ggaattggct
     3121 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
     3181 gggtcggcaa ttgatccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
     3241 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
     3301 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggac cggtgccacc
     3361 atgaaacgga cagccgacgg aagcgagttc gagtcaccaa agaagaagcg gaaagtctct
     3421 gaggtggagt tttcccacga gtactggatg agacatgccc tgaccctggc caagagggca
     3481 cgggatgaga gggaggtgcc tgtgggagcc gtgctggtgc tgaacaatag agtgatcggc
     3541 gagggctgga acagagccat cggcctgcac gacccaacag cccatgccga aattatggcc
     3601 ctgagacagg gcggcctggt catgcagaac tacagactga ttgacgccac cctgtacgtg
     3661 acattcgagc cttgcgtgat gtgcgccggc gccatgatcc actctaggat cggccgcgtg
     3721 gtgtttggcg tgaggaactc aaaaagaggc gccgcaggct ccctgatgaa cgtgctgaac
     3781 taccccggca tgaatcaccg cgtcgaaatt accgagggaa tcctggcaga tgaatgtgcc
     3841 gccctgctgt gcgatttcta tcggatgcct agacaggtgt tcaatgctca gaagaaggcc
     3901 cagagctcca tcaactccgg aggatctagc ggaggctcct ctggctctga gacacctggc
     3961 acaagcgaga gcgcaacacc tgaaagcagc gggggcagca gcggggggtc aatgcaggag
     4021 aaccagcaga agcaaaatta catcctgggc ctggccatcg gcatcaccag cgtgggctat
     4081 ggcctgatcg acagcaagac cagagaagtg attgacgccg gcgtgcggct attcccagag
     4141 gccgactctg aaaacaacag caatagaaga tctaagcggg gcgcccggag actgaaaaga
     4201 cgaagaatcc acagactgaa cagagtgaaa gacctcctgg ctgactacca gatgatcgac
     4261 ttaaacaacg tgcccaagtc taccgacccc tacaccatcc gggtgaaggg actgcgggaa
     4321 cctctgacca aggaagagtt tgccatcgct ctgctgcata tcgccaagag aagaggcctg
     4381 cacaacatct ccgtgagcat gggcgatgag gaacaggaca acgagctgtc caccaagcag
     4441 cagctgcaga agaacgctca gcagctgcag gacaaatacg tgtgcgagct gcaactggaa
     4501 agactgacca acatcaacaa ggtcagaggc gagaagaacc ggttcaagac cgaagatttc
     4561 gtgaaggaag tgaagcagct gtgcgagacc cagcggcagt accacaacat cgacgatcag
     4621 ttcatccagc agtacatcga cctggtgagc acccggagag aatacttcga gggccctggc
     4681 aacggctctc catatggctg ggatggagat ctgctgaagt ggtatgagaa gctgatgggc
     4741 agatgcacct acttccctga ggagctgaga agcgtgaagt acgcctacag cgccgatctg
     4801 tttaacgccc tgaacgatct gaacaacctg gttgtgaccc gcgacgacaa ccctaagctg
     4861 gaatactacg agaaatacca cattatcgag aacgtgttca agcagaagaa aaatccaact
     4921 ctgaagcaaa tcgccaaaga gatcggcgtg caggattacg acatcagagg atacagaatt
     4981 accaagtccg gtaagcctca gttcaccagc ttcaaactct accacgacct gaaaaacatc
     5041 tttgaacagg ccaaatacct ggaagatgtg gagatgctgg atgagatagc taaaatcctg
     5101 acaatctacc aagacgagat cagcatcaag aaagccctgg accagctgcc tgagctgctg
     5161 accgagagcg aaaaaagcca gatcgctcag ctgaccggct acaccggtac acatagactg
     5221 tctctgaagt gcatccacat cgtgatcgac gagctgtggg agagccccga aaaccagatg
     5281 gaaatcttca ccagactgaa cctcaagcca aagaaggtgg aaatgagcga gatcgacagc
     5341 atccctacca cactggtgga tgagttcatc ctgagccctg tggtgaagcg ggccttcatc
     5401 cagtccatca aggtgatcaa cgctgtgatc aacagattcg gcctgcccga ggacatcatt
     5461 atcgagctgg ccagagagaa gaacagcaag gacagaagga agttcatcaa caagctgcag
     5521 aaacagaatg aggccacccg gaaaaaaatc gagcagctgc tggccaagta cggcaatacc
     5581 aatgccaagt acatgatcga aaagatcaag ctgcatgaca tgcaggaggg caagtgtctg
     5641 tacagcctgg aagctatccc cctggaagac ctgctgtcta atcctacaca ctacgaggtg
     5701 gaccacatca tccctagaag cgtgtccttc gacaacagcc tgaacaacaa ggttctggtg
     5761 aagcaaagcg agaacagcaa gaagggcaat aggacccctt accagtacct gagcagcaac
     5821 gagtccaaga tctcttacaa ccagttcaag cagcacattc tgaacctgtc taaggccaaa
     5881 gatagaatca gcaagaagaa acgagatatg ctgctggaag aacgggacat caacaaattc
     5941 gaggtgcaga aggaattcat caacagaaac cttgtggaca cccggtacgc cactcgggag
     6001 ctgagcaacc tgctgaagac ctacttcagc acacacgact acgccgtgaa agtgaagacc
     6061 atcaacggcg gcttcacaaa ccacctgagg aaggtgtggg acttcaagaa gcaccggaac
     6121 cacggctaca agcaccacgc cgaggatgcc ctggtcatcg ccaacgccga ctttctgttc
     6181 aaaacccaca aggccctgag acggacagac aagatcctgg aacagcctgg actggaagtc
     6241 aacgacacca ccgtgaaggt ggacacagag gagaagtacc aggagttatt cgagacaccg
     6301 aaacaagtga agaacatcaa gcagtttaga gatttcaagt attctcacag agttgacaag
     6361 aagcccaacc ggcagctgat caatgatacc ctgtacagta ccagagagat cgatggcgaa
     6421 acctacgtgg tccaaacact gaaagacctg tacgccaagg acaatgaaaa ggtgaaaaag
     6481 ctctttacag aacggcctca aaagatactg atgtaccagc acgatcctaa gacctttgag
     6541 aaactgatga ccattctgaa tcagtacgct gaggcaaaga atcctctggc cgcttattac
     6601 gaggataagg gcgaatacgt gaccaagtac gccaagaagg gcaacggccc tgccatccac
     6661 aagatcaaat acatcgacaa gaaactgggc agctacctgg acgtgagtaa caaatatcct
     6721 gagacacaga acaagctggt gaaactgtct ctgaagagct ttagattcga catctacaaa
     6781 tgtgaacagg gctacaagat ggtgtccatt ggctacctcg acgtactgaa gaaggacaac
     6841 tactactaca tccccaaaga taagtacgag gccgagaagc agaaaaagaa gatcaaggaa
     6901 agcgacctct tcgtgggcag cttctactac aacgacctga tcatgtacga ggacgaactc
     6961 ttccgggtga tcggagtgaa ctccgatatc aacaacctgg ttgagctgaa tatggtcgac
     7021 atcacctaca aggatttctg cgaggtgaac aacgtgacag gcgagaagag aatcaagaaa
     7081 accatcggaa agagagtggt gctgatcgag aagtatacga ccgacatcct gggaaatctg
     7141 tataaaacgc ccctgcctaa gaagccccag ctcattttca agagaggcga gctgtctggc
     7201 ggctcaaaaa gaaccgccga cggcagcgaa ttcgagccca agaagaagag gaaagtctaa
     7261 tagatctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     7321 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     7381 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     7441 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggctcga
     7501 gcggcccaag cttaaaaaaa tctcgccaac aagttgacga gataaacacg gcattttgcc
     7561 ttgttttagt agattctgta attttcatta cagagtacta aaaccaggca tggtttggta
     7621 agtagcggtg tttcgtcctt tccacaagat atataaagcc aagaaatcga aatactttca
     7681 agttacggta agcatatgat agtccatttt aaaacataat tttaaaactg caaactaccc
     7741 aagaaattat tactttctac gtcacgtatt ttgtactaat atctttgtgt ttacagtcaa
     7801 attaattcta attatctctc taacagcctt gtatcgtata tgcaaatatg aaggaatcat
     7861 gggaaatagg ccctcttcct gcccgacctt gcggccgc
//
