LOCUS       sequence-411194-        7255 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SpG cas9C by the constitutive CASI 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 7255)
  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 7255)
  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-CASI-InteinC-SpG C aa714-1368-U6-Lmna sgRNA1
FEATURES             Location/Qualifiers
     source          1..7255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        40..317
                     /label=chicken beta-actin promoter
     regulatory      791..800
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      795..804
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2960..2980
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2995..3050
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3049..3068)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     misc_RNA        complement(3155..3230)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(3251..3499)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(3309..3328)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(3479..3499)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     repeat_region   3834..3963
                     /label=AAV2 ITR
     primer_bind     complement(3982..3999)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3982..3998)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3991..4013)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4140..4595
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4227..4246)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4437..4458
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4621..4725
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4726..4794,4795..5586)
                     /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(4944..4963)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5757..6345
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6246..6265
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6633..6654
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6669..6674,6675..6692,6693..6699)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6707..6723
                     /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     6712..6734
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6731..6747
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6731..6747
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   6785..6914
                     /label=AAV2 ITR
     enhancer        join(6990..7255,1..38)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct
       61 gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt
      121 aattattttg tgcagcgatg ggggcggggg gggggggggg gsgcgcgcsa gggggggggg
      181 gggggggsga gggggggggs ggggsgaggc ggagaggtgc ggcggcagcc aatcagagcg
      241 gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc
      301 gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg
      361 ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actaaaacag gtaagtccgg
      421 cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc tcctcacggc gagcgctgcc
      481 acgtcagacg aagggcgcag cgagcgtcct gatccttccg cccggacgct caggacagcg
      541 gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga cattttagga
      601 cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac aggcgaggaa
      661 aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga acgccgatga
      721 tgcctctact aaccatgttc atgttttctt tttttttcta caggtcctgg gtgacgaaca
      781 ggctagcgcc gccaccatgg cggcggcgtg cccggaactg cgtcagctgg cgcagagcga
      841 tgtgtattgg gatccgattg tgagcattga accggatggc gtggaagaag tgtttgatct
      901 gaccgtgccg ggcccgcata actttgtggc gaacgatatt attgcgcata actctggcca
      961 gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg
     1021 catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc
     1081 cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa
     1141 cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct
     1201 gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct
     1261 gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta
     1321 cgatgtggac catatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt
     1381 gctgaccaga agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt
     1441 gaagaagatg aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa
     1501 gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt
     1561 catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga
     1621 ctcccggatg aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat
     1681 caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg
     1741 cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc
     1801 cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta
     1861 cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta
     1921 cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga
     1981 gatccggaag cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa
     2041 gggccgggat tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa
     2101 aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag
     2161 cgataagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcctgtg
     2221 gcccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa
     2281 actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa
     2341 gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat
     2401 caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc
     2461 tgccaagcag ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct
     2521 gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca
     2581 gctgtttgtg gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt
     2641 ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa
     2701 gcaccgggat aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac
     2761 caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagcagta
     2821 cagaagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta
     2881 cgagacacgg atcgacctgt ctcagctggg aggtgactct ggcggctcaa aaagaaccgc
     2941 cgacggcagc gaattcgagc ccaagaagaa gaggaaagtc taaaccggtt ggctaataaa
     3001 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga
     3061 catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa
     3121 catatgccca tatgaggccc tcacgcgtaa aaaagcaccg actcggtgcc actttttcaa
     3181 gttgataacg gactagcctt attttaactt gctatttcta gctctaaaac ggaagtggcc
     3241 atgtgcaagc ggtgtttcgt cctttccaca agatatataa agccaagaaa tcgaaatact
     3301 ttcaagttac ggtaagcata tgatagtcca ttttaaaaca taattttaaa actgcaaact
     3361 acccaagaaa ttattacttt ctacgtcacg tattttgtac taatatcttt gtgtttacag
     3421 tcaaattaat tccaattatc tctctaacag ccttgtatcg tatatgcaaa tatgaaggaa
     3481 tcatgggaaa taggccctcg gtaccggcga tgctggctgc catgaacaaa ggttggctat
     3541 aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag
     3601 ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat
     3661 ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc
     3721 cagtcatagc tgtccctctt ctcttatgga gatcggatcc gaattcccga taaggatctt
     3781 cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc
     3841 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     3901 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     3961 cagccttaat taacctaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     4021 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     4081 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     4141 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4201 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4261 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4321 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4381 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4441 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4501 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4561 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     4621 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4681 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4741 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4801 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4861 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4921 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     4981 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5041 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5101 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5161 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5221 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5281 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5341 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5401 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5461 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5521 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5581 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5641 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5701 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5761 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5821 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5881 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5941 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6001 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6061 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6121 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6181 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6241 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6301 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6361 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6421 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     6481 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     6541 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     6601 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     6661 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     6721 caatttcaca caggaaacag ctatgaccat gattacgcca gatttaatta aggccttaat
     6781 taggctgcgc gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac
     6841 ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat
     6901 cactaggggt tccttgtagt taatgattaa cccgccatgc tacttatcta cgtagccatg
     6961 ctctaggaag atcggaattc ggagttccgc gttacataac ttacggtaaa tggcccgcct
     7021 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     7081 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     7141 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     7201 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tactt
//
