LOCUS       sequence-212733-        7048 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV inverted terminal repeat based vector plasmid encoding E. coli 
            TadA, the N-terminal half of nCas9 and sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7048)
  AUTHORS   Ryu SM, Koo T, Kim K, Lim K, Baek G, Kim ST, Kim HS, Kim DE, Lee H, 
            Chung E, Kim JS
  TITLE     Adenine base editing in mouse embryos and an adult mouse model of 
            Duchenne muscular dystrophy.
  JOURNAL   Nat Biotechnol. 2018 Jul;36(6):536-539. doi: 10.1038/nbt.4148. Epub 
            2018 Apr 27.
  PUBMED    29702637
REFERENCE   2  (bases 1 to 7048)
  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"; doi: "10.1038/nbt.4148"; 
            journalName: "Nat Biotechnol"; date: "2018-07"; volume: "36"; issue:
            "6"; pages: "536-539"
COMMENT     Sequence Label: pAAV-ABE-NT-sgRNA
FEATURES             Location/Qualifiers
     source          1..7048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     primer_bind     312..331
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        411..486
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     846..862
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     846..862
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             885..905
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     repeat_region   4311..4451
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4311..4440
                     /label=AAV2 ITR
     rep_origin      4526..4981
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4613..4632)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4823..4844
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4998..5017)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5117..5139
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5177..5195)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5263..5367
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5368..5436,5437..6228)
                     /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(5586..5605)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6399..6987
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6888..6907
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtggagct ttcccatgat tccttcatat ttgcatatac
      181 gatacaaggc tgttagagag ataattggaa ttaatttgac tgtaaacaca aagatattag
      241 tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat
      301 gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt
      361 tatatatctt gtggaaagga cgaaacaccg aactagcttt taattgctgt gttttagagc
      421 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
      481 cggtgctttt ttctagattc gtagccaccg cggtggcggc cgtccgccct cggcaccatc
      541 ctcacgacac ccaaatatgg cgacgggtga ggaatggtgg ggagttattt ttagagcggt
      601 gaggaaggtg ggcaggcagc aggtgttggc gctctaaaaa taactcccgg gagttatttt
      661 tagagcggag gaatggtgga cacccaaata tggcgacggt tcctcacccg tcgccatatt
      721 tgggtgtccg ccctcggccg gggccgcatt cctgggggcc gggcggtgct cccgcccgcc
      781 tcgataaaag gctccggggc cggcggcggc ccacgagcta cccggaggag cgggaggcgc
      841 caagctctag aactagtgga tcccccaagc ttgccgccac catgccaaaa aagaagagaa
      901 aggtagaagc gagtgaagtg gaattctccc acgagtattg gatgagacac gcattgacgc
      961 tcgcaaaaag agcgtgggac gagagggaag tacccgtcgg agcagtcctc gtgcataaca
     1021 atcgggtgat aggcgagggc tggaatcgac ctatcggccg acatgacccg accgcccatg
     1081 cagaaattat ggctctgcga cagggagggt tggtgatgca gaactatcgg cttatcgatg
     1141 ccacactcta cgttacgctt gaaccgtgcg ttatgtgtgc gggggcgatg atacatagta
     1201 gaatagggag ggtcgtgttc ggggcgcgag atgcgaaaac cggggccgca ggctcattga
     1261 tggacgtcct gcaccacccc ggcatgaacc atcgggtcga gataaccgaa ggcatattgg
     1321 cagacgaatg tgctgcgctc ctttccgatt tctttaggat gcgacgccag gagatcaagg
     1381 cgcaaaaaaa agcccagtct agtaccgact ccgggggcag cagtgggggg tctagtggca
     1441 gtgagacgcc cggtacgagc gagagcgcca cgcctgaatc atcaggtgga agcagtggtg
     1501 gctcttctga agtcgaattt tcccacgagt actggatgag acacgcactt acgcttgcca
     1561 agagggcaag agatgaacgc gaggtcccgg tcggagcagt gcttgtcctt aataatagag
     1621 tgataggcga aggctggaat cgagctatcg gcctccacga tcctaccgcg catgctgaga
     1681 tcatggcgct gcgacaaggg gggctggtaa tgcaaaacta tagactcatc gatgcgacgc
     1741 tgtatgtgac ctttgagccc tgtgttatgt gtgctggggc tatgatacat agcagaattg
     1801 gcagggtagt cttcggcgtc cgcaatgcca aaaccggggc agctggatcc ttgatggatg
     1861 ttttgcatta ccctggaatg aatcataggg ttgagattac agaaggtatc ttggcggatg
     1921 aatgcgccgc acttctctgt tatttttttc ggatgccgag gcaagtcttc aatgcccaga
     1981 aaaaagcgca gtctagtaca gacagtggag gctcaagtgg gggcagctca ggaagcgaaa
     2041 ctcctggtac ctcagagtcc gctactcccg aatcctctgg gggtagctca gggggcagta
     2101 tggacaagaa gtacagcatc ggcctggcca tcggtaccaa cagcgtgggc tgggccgtga
     2161 tcaccgacga gtacaaggtg cccagcaaga agttcaaggt gctgggcaac accgaccgcc
     2221 acagcatcaa gaagaacctg atcggcgccc tgctgttcga cagcggcgag accgccgagg
     2281 ccacccgcct gaagcgcacc gcccgccgcc gctacacccg ccgcaagaac cgcatctgct
     2341 acctgcagga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccaccgcc
     2401 tggaggagag cttcctggtg gaggaggaca agaagcacga gcgccacccc atcttcggca
     2461 acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac ctgcgcaaga
     2521 agctggtgga cagcaccgac aaggccgacc tgcgcctgat ctacctggcc ctggcccaca
     2581 tgatcaagtt ccgcggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg
     2641 tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag gagaacccca
     2701 tcaacgccag cggcgtggac gccaaggcca tcctgagcgc ccgcctgagc aagagccgcc
     2761 gcctggagaa cctgatcgcc cagctgcccg gcgagaagaa gaacggcctg ttcggcaacc
     2821 tgatcgccct gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg
     2881 acgccaagct gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc
     2941 agatcggcga ccagtacgcc gacctgttcc tggccgccaa gaacctgagc gacgccatcc
     3001 tgctgagcga catcctgcgc gtgaacaccg agatcaccaa ggcccccctg agcgccagca
     3061 tgatcaagcg ctacgacgag caccaccagg acctgaccct gctgaaggcc ctggtgcgcc
     3121 agcagctgcc cgagaagtac aaggagatct tcttcgacca gagcaagaac ggctacgccg
     3181 gctacatcga cggcggcgcc agccaggagg agttctacaa gttcatcaag cccatcctgg
     3241 agaagatgga cggcaccgag gagctgctgg tgaagctgaa ccgcgaggac ctgctgcgca
     3301 agcagcgcac cttcgacaac ggcagcatcc cccaccagat ccacctgggc gagctgcacg
     3361 ccatcctgcg ccgccaggag gacttctacc ccttcctgaa ggacaaccgc gagaagatcg
     3421 agaagatcct gaccttccgc atcccctact acgtgggccc cctggcccgc ggcaacagcc
     3481 gcttcgcctg gatgacccgc aagagcgagg agaccatcac cccctggaac ttcgaggagg
     3541 tggtggacaa gggcgccagc gcccagagct tcatcgagcg catgaccaac ttcgacaaga
     3601 acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt
     3661 acaacgagct gaccaaggtg aagtacgtga ccgagggcat gcgcaagccc gccttcctga
     3721 gcggcgagca gaagaaggcc atcgtggacc tgctgttcaa gaccaaccgc aaggtgaccg
     3781 tgaagcagct gaaggaggac tacttcaaga agatcgagtg cttcgacagc gtggagatca
     3841 gcggcgtgga ggaccgcttc aacgccagcc tgggcaccta ccacgacctg ctgaagatca
     3901 tcaaggacaa ggacttcctg gacaacgagg agaacgagga catcctggag gacatcgtgc
     3961 tgaccctgac cctgttcgag gaccgcgaga tgatcgagga gcgcctgaag acctacgccc
     4021 acctgttcga cgacaaggtg atgaagcagc tgaagcgccg ccgctacacc ggctggggcc
     4081 gcctgagccg caagcttatc aacggcatcc gcgacaagca gagcggcaag accatcctgg
     4141 acttcctgaa gagcgacggc ttcgccaacc gcaacttcat gcagctgatc cacgacgaca
     4201 gcctgacctt caaggaggac atccagaagg cccaggtgag cggcgtaagt atcaaggtta
     4261 caagacaggt ttaaggagac caatagaaac tgggcggacc gagcggccgc aggaacccct
     4321 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     4381 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     4441 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4501 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4561 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4621 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4681 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4741 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4801 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4861 atctcgggct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4921 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     4981 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     5041 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     5101 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     5161 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     5221 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     5281 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     5341 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     5401 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     5461 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     5521 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     5581 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     5641 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5701 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5761 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5821 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5881 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5941 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6001 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6061 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6121 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6181 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     6241 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6301 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6361 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6421 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6481 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6541 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6601 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6661 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6721 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6781 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6841 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6901 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6961 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7021 gccttttgct ggccttttgc tcacatgt
//
