LOCUS       sequence-436701-        8021 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses SauriABE8e by MYL2 promoter and sgRNA targeting murine 
            Camk2d intron6 5' splice site by U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8021)
  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 8021)
  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-MYL2-TadA8e-Sauri-U6-Camk2d sgRNA6
FEATURES             Location/Qualifiers
     source          1..8021
                     /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
     CDS             3520..3540
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7360..7380
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    7394..7618
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(7750..7998)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(7808..7827)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(7978..7998)
                     /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 tcagtgggtt actcttcccc attagacaat ggcagggaag agagcacacc
     3181 ccatcatccc caggccaggc cccagccact gactctttaa ccttgaaggc atttttgggt
     3241 ctcacgtgtc cacccaggcg ggtggccgcc tttgagcagc tcttacttca gaagaacggc
     3301 atggagtggg gggtgggggg cttaggtggc ctccgcctca cctacaactg ccaaaagtgg
     3361 tcatggggtt atttttaacc ccaggggaga ggtatttatt gttccacagc aggggcagag
     3421 gccagcaggc tcctcgaact ctccagaggt ggcaactggc ctcagacacc gaccggtgcc
     3481 accatgaaac ggacagccga cggaagcgag ttcgagtcac caaagaagaa gcggaaagtc
     3541 tctgaggtgg agttttccca cgagtactgg atgagacatg ccctgaccct ggccaagagg
     3601 gcacgggatg agagggaggt gcctgtggga gccgtgctgg tgctgaacaa tagagtgatc
     3661 ggcgagggct ggaacagagc catcggcctg cacgacccaa cagcccatgc cgaaattatg
     3721 gccctgagac agggcggcct ggtcatgcag aactacagac tgattgacgc caccctgtac
     3781 gtgacattcg agccttgcgt gatgtgcgcc ggcgccatga tccactctag gatcggccgc
     3841 gtggtgtttg gcgtgaggaa ctcaaaaaga ggcgccgcag gctccctgat gaacgtgctg
     3901 aactaccccg gcatgaatca ccgcgtcgaa attaccgagg gaatcctggc agatgaatgt
     3961 gccgccctgc tgtgcgattt ctatcggatg cctagacagg tgttcaatgc tcagaagaag
     4021 gcccagagct ccatcaactc cggaggatct agcggaggct cctctggctc tgagacacct
     4081 ggcacaagcg agagcgcaac acctgaaagc agcgggggca gcagcggggg gtcaatgcag
     4141 gagaaccagc agaagcaaaa ttacatcctg ggcctggcca tcggcatcac cagcgtgggc
     4201 tatggcctga tcgacagcaa gaccagagaa gtgattgacg ccggcgtgcg gctattccca
     4261 gaggccgact ctgaaaacaa cagcaataga agatctaagc ggggcgcccg gagactgaaa
     4321 agacgaagaa tccacagact gaacagagtg aaagacctcc tggctgacta ccagatgatc
     4381 gacttaaaca acgtgcccaa gtctaccgac ccctacacca tccgggtgaa gggactgcgg
     4441 gaacctctga ccaaggaaga gtttgccatc gctctgctgc atatcgccaa gagaagaggc
     4501 ctgcacaaca tctccgtgag catgggcgat gaggaacagg acaacgagct gtccaccaag
     4561 cagcagctgc agaagaacgc tcagcagctg caggacaaat acgtgtgcga gctgcaactg
     4621 gaaagactga ccaacatcaa caaggtcaga ggcgagaaga accggttcaa gaccgaagat
     4681 ttcgtgaagg aagtgaagca gctgtgcgag acccagcggc agtaccacaa catcgacgat
     4741 cagttcatcc agcagtacat cgacctggtg agcacccgga gagaatactt cgagggccct
     4801 ggcaacggct ctccatatgg ctgggatgga gatctgctga agtggtatga gaagctgatg
     4861 ggcagatgca cctacttccc tgaggagctg agaagcgtga agtacgccta cagcgccgat
     4921 ctgtttaacg ccctgaacga tctgaacaac ctggttgtga cccgcgacga caaccctaag
     4981 ctggaatact acgagaaata ccacattatc gagaacgtgt tcaagcagaa gaaaaatcca
     5041 actctgaagc aaatcgccaa agagatcggc gtgcaggatt acgacatcag aggatacaga
     5101 attaccaagt ccggtaagcc tcagttcacc agcttcaaac tctaccacga cctgaaaaac
     5161 atctttgaac aggccaaata cctggaagat gtggagatgc tggatgagat agctaaaatc
     5221 ctgacaatct accaagacga gatcagcatc aagaaagccc tggaccagct gcctgagctg
     5281 ctgaccgaga gcgaaaaaag ccagatcgct cagctgaccg gctacaccgg tacacataga
     5341 ctgtctctga agtgcatcca catcgtgatc gacgagctgt gggagagccc cgaaaaccag
     5401 atggaaatct tcaccagact gaacctcaag ccaaagaagg tggaaatgag cgagatcgac
     5461 agcatcccta ccacactggt ggatgagttc atcctgagcc ctgtggtgaa gcgggccttc
     5521 atccagtcca tcaaggtgat caacgctgtg atcaacagat tcggcctgcc cgaggacatc
     5581 attatcgagc tggccagaga gaagaacagc aaggacagaa ggaagttcat caacaagctg
     5641 cagaaacaga atgaggccac ccggaaaaaa atcgagcagc tgctggccaa gtacggcaat
     5701 accaatgcca agtacatgat cgaaaagatc aagctgcatg acatgcagga gggcaagtgt
     5761 ctgtacagcc tggaagctat ccccctggaa gacctgctgt ctaatcctac acactacgag
     5821 gtggaccaca tcatccctag aagcgtgtcc ttcgacaaca gcctgaacaa caaggttctg
     5881 gtgaagcaaa gcgagaacag caagaagggc aataggaccc cttaccagta cctgagcagc
     5941 aacgagtcca agatctctta caaccagttc aagcagcaca ttctgaacct gtctaaggcc
     6001 aaagatagaa tcagcaagaa gaaacgagat atgctgctgg aagaacggga catcaacaaa
     6061 ttcgaggtgc agaaggaatt catcaacaga aaccttgtgg acacccggta cgccactcgg
     6121 gagctgagca acctgctgaa gacctacttc agcacacacg actacgccgt gaaagtgaag
     6181 accatcaacg gcggcttcac aaaccacctg aggaaggtgt gggacttcaa gaagcaccgg
     6241 aaccacggct acaagcacca cgccgaggat gccctggtca tcgccaacgc cgactttctg
     6301 ttcaaaaccc acaaggccct gagacggaca gacaagatcc tggaacagcc tggactggaa
     6361 gtcaacgaca ccaccgtgaa ggtggacaca gaggagaagt accaggagtt attcgagaca
     6421 ccgaaacaag tgaagaacat caagcagttt agagatttca agtattctca cagagttgac
     6481 aagaagccca accggcagct gatcaatgat accctgtaca gtaccagaga gatcgatggc
     6541 gaaacctacg tggtccaaac actgaaagac ctgtacgcca aggacaatga aaaggtgaaa
     6601 aagctcttta cagaacggcc tcaaaagata ctgatgtacc agcacgatcc taagaccttt
     6661 gagaaactga tgaccattct gaatcagtac gctgaggcaa agaatcctct ggccgcttat
     6721 tacgaggata agggcgaata cgtgaccaag tacgccaaga agggcaacgg ccctgccatc
     6781 cacaagatca aatacatcga caagaaactg ggcagctacc tggacgtgag taacaaatat
     6841 cctgagacac agaacaagct ggtgaaactg tctctgaaga gctttagatt cgacatctac
     6901 aaatgtgaac agggctacaa gatggtgtcc attggctacc tcgacgtact gaagaaggac
     6961 aactactact acatccccaa agataagtac gaggccgaga agcagaaaaa gaagatcaag
     7021 gaaagcgacc tcttcgtggg cagcttctac tacaacgacc tgatcatgta cgaggacgaa
     7081 ctcttccggg tgatcggagt gaactccgat atcaacaacc tggttgagct gaatatggtc
     7141 gacatcacct acaaggattt ctgcgaggtg aacaacgtga caggcgagaa gagaatcaag
     7201 aaaaccatcg gaaagagagt ggtgctgatc gagaagtata cgaccgacat cctgggaaat
     7261 ctgtataaaa cgcccctgcc taagaagccc cagctcattt tcaagagagg cgagctgtct
     7321 ggcggctcaa aaagaaccgc cgacggcagc gaattcgagc ccaagaagaa gaggaaagtc
     7381 taatagatct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     7441 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     7501 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     7561 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     7621 cgagcggccc aagcttaaaa aaatctcgcc aacaagttga cgagataaac acggcatttt
     7681 gccttgtttt agtagattct gtaattttca ttacagagta ctaaaaccgg gacctaaagg
     7741 tgagtaaacg gtgtttcgtc ctttccacaa gatatataaa gccaagaaat cgaaatactt
     7801 tcaagttacg gtaagcatat gatagtccat tttaaaacat aattttaaaa ctgcaaacta
     7861 cccaagaaat tattactttc tacgtcacgt attttgtact aatatctttg tgtttacagt
     7921 caaattaatt ctaattatct ctctaacagc cttgtatcgt atatgcaaat atgaaggaat
     7981 catgggaaat aggccctctt cctgcccgac cttgcggccg c
//
