LOCUS       sequence-411205-        7897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SauriABE8e by EFS promoter and sgRNA targeting murine Lmna
            c.1621T mutation by U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7897)
  AUTHORS   Yang L, Liu Z, Sun J, Chen Z, Gao F, Guo Y
  TITLE     Adenine base editor-based correction of the cardiac pathogenic Lmna 
            c.1621C > T mutation in murine hearts.
  JOURNAL   J Cell Mol Med. 2024 Feb;28(4):e18145. doi: 10.1111/jcmm.18145.
  PUBMED    38332517
REFERENCE   2  (bases 1 to 7897)
  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: "J Cell Mol 
            Med."; date: "2024-02"; volume: "28(4):e18145. doi"; pages: " 
            10.1111/jcmm.18145"
COMMENT     Sequence Label: pAAV-EFS-TadA8e-Sauri-U6-Lmna sgRNA
FEATURES             Location/Qualifiers
     source          1..7897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    266..287
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(302..307,308..325,326..332)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    340..356
                     /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     345..367
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     364..380
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     364..380
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   407..536
                     /label=AAV2 ITR
     promoter        602..813
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             863..883
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4703..4723
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4737..4961
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(5092..5340)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(5150..5169)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(5320..5340)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     repeat_region   5364..5493
                     /label=AAV2 ITR
     primer_bind     complement(5512..5529)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5512..5528)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5521..5543)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5670..6125
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5757..5776)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5967..5988
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6151..6255
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6256..6324,6325..7116)
                     /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(6474..6493)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7287..7875
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7776..7795
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
       61 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
      121 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
      181 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
      241 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
      301 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
      361 acacaggaaa cagctatgac catgattacg ccagatttaa ttaaggctgc gcgctcgctc
      421 gctcactgag gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc
      481 agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg
      541 cctctagaat tcgctagcta ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag
      601 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
      661 tccggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
      721 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
      781 ctttttcgca acgggtttgc cgccagaaca caggaccggt gccaccatga aacggacagc
      841 cgacggaagc gagttcgagt caccaaagaa gaagcggaaa gtctctgagg tggagttttc
      901 ccacgagtac tggatgagac atgccctgac cctggccaag agggcacggg atgagaggga
      961 ggtgcctgtg ggagccgtgc tggtgctgaa caatagagtg atcggcgagg gctggaacag
     1021 agccatcggc ctgcacgacc caacagccca tgccgaaatt atggccctga gacagggcgg
     1081 cctggtcatg cagaactaca gactgattga cgccaccctg tacgtgacat tcgagccttg
     1141 cgtgatgtgc gccggcgcca tgatccactc taggatcggc cgcgtggtgt ttggcgtgag
     1201 gaactcaaaa agaggcgccg caggctccct gatgaacgtg ctgaactacc ccggcatgaa
     1261 tcaccgcgtc gaaattaccg agggaatcct ggcagatgaa tgtgccgccc tgctgtgcga
     1321 tttctatcgg atgcctagac aggtgttcaa tgctcagaag aaggcccaga gctccatcaa
     1381 ctccggagga tctagcggag gctcctctgg ctctgagaca cctggcacaa gcgagagcgc
     1441 aacacctgaa agcagcgggg gcagcagcgg ggggtcaatg caggagaacc agcagaagca
     1501 aaattacatc ctgggcctgg ccatcggcat caccagcgtg ggctatggcc tgatcgacag
     1561 caagaccaga gaagtgattg acgccggcgt gcggctattc ccagaggccg actctgaaaa
     1621 caacagcaat agaagatcta agcggggcgc ccggagactg aaaagacgaa gaatccacag
     1681 actgaacaga gtgaaagacc tcctggctga ctaccagatg atcgacttaa acaacgtgcc
     1741 caagtctacc gacccctaca ccatccgggt gaagggactg cgggaacctc tgaccaagga
     1801 agagtttgcc atcgctctgc tgcatatcgc caagagaaga ggcctgcaca acatctccgt
     1861 gagcatgggc gatgaggaac aggacaacga gctgtccacc aagcagcagc tgcagaagaa
     1921 cgctcagcag ctgcaggaca aatacgtgtg cgagctgcaa ctggaaagac tgaccaacat
     1981 caacaaggtc agaggcgaga agaaccggtt caagaccgaa gatttcgtga aggaagtgaa
     2041 gcagctgtgc gagacccagc ggcagtacca caacatcgac gatcagttca tccagcagta
     2101 catcgacctg gtgagcaccc ggagagaata cttcgagggc cctggcaacg gctctccata
     2161 tggctgggat ggagatctgc tgaagtggta tgagaagctg atgggcagat gcacctactt
     2221 ccctgaggag ctgagaagcg tgaagtacgc ctacagcgcc gatctgttta acgccctgaa
     2281 cgatctgaac aacctggttg tgacccgcga cgacaaccct aagctggaat actacgagaa
     2341 ataccacatt atcgagaacg tgttcaagca gaagaaaaat ccaactctga agcaaatcgc
     2401 caaagagatc ggcgtgcagg attacgacat cagaggatac agaattacca agtccggtaa
     2461 gcctcagttc accagcttca aactctacca cgacctgaaa aacatctttg aacaggccaa
     2521 atacctggaa gatgtggaga tgctggatga gatagctaaa atcctgacaa tctaccaaga
     2581 cgagatcagc atcaagaaag ccctggacca gctgcctgag ctgctgaccg agagcgaaaa
     2641 aagccagatc gctcagctga ccggctacac cggtacacat agactgtctc tgaagtgcat
     2701 ccacatcgtg atcgacgagc tgtgggagag ccccgaaaac cagatggaaa tcttcaccag
     2761 actgaacctc aagccaaaga aggtggaaat gagcgagatc gacagcatcc ctaccacact
     2821 ggtggatgag ttcatcctga gccctgtggt gaagcgggcc ttcatccagt ccatcaaggt
     2881 gatcaacgct gtgatcaaca gattcggcct gcccgaggac atcattatcg agctggccag
     2941 agagaagaac agcaaggaca gaaggaagtt catcaacaag ctgcagaaac agaatgaggc
     3001 cacccggaaa aaaatcgagc agctgctggc caagtacggc aataccaatg ccaagtacat
     3061 gatcgaaaag atcaagctgc atgacatgca ggagggcaag tgtctgtaca gcctggaagc
     3121 tatccccctg gaagacctgc tgtctaatcc tacacactac gaggtggacc acatcatccc
     3181 tagaagcgtg tccttcgaca acagcctgaa caacaaggtt ctggtgaagc aaagcgagaa
     3241 cagcaagaag ggcaatagga ccccttacca gtacctgagc agcaacgagt ccaagatctc
     3301 ttacaaccag ttcaagcagc acattctgaa cctgtctaag gccaaagata gaatcagcaa
     3361 gaagaaacga gatatgctgc tggaagaacg ggacatcaac aaattcgagg tgcagaagga
     3421 attcatcaac agaaaccttg tggacacccg gtacgccact cgggagctga gcaacctgct
     3481 gaagacctac ttcagcacac acgactacgc cgtgaaagtg aagaccatca acggcggctt
     3541 cacaaaccac ctgaggaagg tgtgggactt caagaagcac cggaaccacg gctacaagca
     3601 ccacgccgag gatgccctgg tcatcgccaa cgccgacttt ctgttcaaaa cccacaaggc
     3661 cctgagacgg acagacaaga tcctggaaca gcctggactg gaagtcaacg acaccaccgt
     3721 gaaggtggac acagaggaga agtaccagga gttattcgag acaccgaaac aagtgaagaa
     3781 catcaagcag tttagagatt tcaagtattc tcacagagtt gacaagaagc ccaaccggca
     3841 gctgatcaat gataccctgt acagtaccag agagatcgat ggcgaaacct acgtggtcca
     3901 aacactgaaa gacctgtacg ccaaggacaa tgaaaaggtg aaaaagctct ttacagaacg
     3961 gcctcaaaag atactgatgt accagcacga tcctaagacc tttgagaaac tgatgaccat
     4021 tctgaatcag tacgctgagg caaagaatcc tctggccgct tattacgagg ataagggcga
     4081 atacgtgacc aagtacgcca agaagggcaa cggccctgcc atccacaaga tcaaatacat
     4141 cgacaagaaa ctgggcagct acctggacgt gagtaacaaa tatcctgaga cacagaacaa
     4201 gctggtgaaa ctgtctctga agagctttag attcgacatc tacaaatgtg aacagggcta
     4261 caagatggtg tccattggct acctcgacgt actgaagaag gacaactact actacatccc
     4321 caaagataag tacgaggccg agaagcagaa aaagaagatc aaggaaagcg acctcttcgt
     4381 gggcagcttc tactacaacg acctgatcat gtacgaggac gaactcttcc gggtgatcgg
     4441 agtgaactcc gatatcaaca acctggttga gctgaatatg gtcgacatca cctacaagga
     4501 tttctgcgag gtgaacaacg tgacaggcga gaagagaatc aagaaaacca tcggaaagag
     4561 agtggtgctg atcgagaagt atacgaccga catcctggga aatctgtata aaacgcccct
     4621 gcctaagaag ccccagctca ttttcaagag aggcgagctg tctggcggct caaaaagaac
     4681 cgccgacggc agcgaattcg agcccaagaa gaagaggaaa gtctaataga tctcgactgt
     4741 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     4801 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     4861 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     4921 agacaatagc aggcatgctg gggatgcggt gggctctatg gctcgagcgg cccaagctta
     4981 aaaaaatctc gccaacaagt tgacgagata aacacggcat tttgccttgt tttagtagat
     5041 tctgtaattt tcattacaga gtactaaaac aggaagtggc catgtgcaag cggtgtttcg
     5101 tcctttccac aagatatata aagccaagaa atcgaaatac tttcaagtta cggtaagcat
     5161 atgatagtcc attttaaaac ataattttaa aactgcaaac tacccaagaa attattactt
     5221 tctacgtcac gtattttgta ctaatatctt tgtgtttaca gtcaaattaa ttctaattat
     5281 ctctctaaca gccttgtatc gtatatgcaa atatgaagga atcatgggaa ataggccctc
     5341 ttcctgcccg accttgcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     5401 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     5461 ccgggcggcc tcagtgagcg agcgagcgcg cagccttaat taacctaatt cactggccgt
     5521 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     5581 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     5641 acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     5701 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     5761 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     5821 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     5881 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     5941 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     6001 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     6061 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     6121 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     6181 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     6241 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     6301 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     6361 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     6421 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     6481 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     6541 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     6601 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     6661 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     6721 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6781 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     6841 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     6901 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6961 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     7021 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     7081 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     7141 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     7201 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     7261 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     7321 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     7381 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     7441 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     7501 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     7561 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     7621 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     7681 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     7741 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7801 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     7861 cggagcctat ggaaaaacgc cagcaacgcg gcctttt
//
