LOCUS       Exported                7052 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV inverted terminal repeat based vector plasmid encoding E. coli 
            TadA, the N-terminal half of nCas9 and sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-ABE-NT-sgRNA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7052)
  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 7052)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7052
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        145..385
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     145..165
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     316..335
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        415..490
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     850..866
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     850..866
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             889..909
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     repeat_region   4315..4455
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4315..4444
                     /label=AAV2 ITR
     rep_origin      4530..4985
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4617..4636)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4827..4848
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5002..5021)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5121..5143
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5181..5199)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5267..5371
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5372..6232
                     /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(5590..5609)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6403..6991
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6892..6911
                     /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 gcgtgagggc ctatttccca tgattccttc atatttgcat
      181 atacgataca aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata
      241 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
      301 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
      361 gctttatata tcttgtggaa aggacgaaac accgaactag cttttaattg ctgtgtttta
      421 gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa aagtggcacc
      481 gagtcggtgc ttttttctag attcgtagcc accgcggtgg cggccgtccg ccctcggcac
      541 catcctcacg acacccaaat atggcgacgg gtgaggaatg gtggggagtt atttttagag
      601 cggtgaggaa ggtgggcagg cagcaggtgt tggcgctcta aaaataactc ccgggagtta
      661 tttttagagc ggaggaatgg tggacaccca aatatggcga cggttcctca cccgtcgcca
      721 tatttgggtg tccgccctcg gccggggccg cattcctggg ggccgggcgg tgctcccgcc
      781 cgcctcgata aaaggctccg gggccggcgg cggcccacga gctacccgga ggagcgggag
      841 gcgccaagct ctagaactag tggatccccc aagcttgccg ccaccatgcc aaaaaagaag
      901 agaaaggtag aagcgagtga agtggaattc tcccacgagt attggatgag acacgcattg
      961 acgctcgcaa aaagagcgtg ggacgagagg gaagtacccg tcggagcagt cctcgtgcat
     1021 aacaatcggg tgataggcga gggctggaat cgacctatcg gccgacatga cccgaccgcc
     1081 catgcagaaa ttatggctct gcgacaggga gggttggtga tgcagaacta tcggcttatc
     1141 gatgccacac tctacgttac gcttgaaccg tgcgttatgt gtgcgggggc gatgatacat
     1201 agtagaatag ggagggtcgt gttcggggcg cgagatgcga aaaccggggc cgcaggctca
     1261 ttgatggacg tcctgcacca ccccggcatg aaccatcggg tcgagataac cgaaggcata
     1321 ttggcagacg aatgtgctgc gctcctttcc gatttcttta ggatgcgacg ccaggagatc
     1381 aaggcgcaaa aaaaagccca gtctagtacc gactccgggg gcagcagtgg ggggtctagt
     1441 ggcagtgaga cgcccggtac gagcgagagc gccacgcctg aatcatcagg tggaagcagt
     1501 ggtggctctt ctgaagtcga attttcccac gagtactgga tgagacacgc acttacgctt
     1561 gccaagaggg caagagatga acgcgaggtc ccggtcggag cagtgcttgt ccttaataat
     1621 agagtgatag gcgaaggctg gaatcgagct atcggcctcc acgatcctac cgcgcatgct
     1681 gagatcatgg cgctgcgaca aggggggctg gtaatgcaaa actatagact catcgatgcg
     1741 acgctgtatg tgacctttga gccctgtgtt atgtgtgctg gggctatgat acatagcaga
     1801 attggcaggg tagtcttcgg cgtccgcaat gccaaaaccg gggcagctgg atccttgatg
     1861 gatgttttgc attaccctgg aatgaatcat agggttgaga ttacagaagg tatcttggcg
     1921 gatgaatgcg ccgcacttct ctgttatttt tttcggatgc cgaggcaagt cttcaatgcc
     1981 cagaaaaaag cgcagtctag tacagacagt ggaggctcaa gtgggggcag ctcaggaagc
     2041 gaaactcctg gtacctcaga gtccgctact cccgaatcct ctgggggtag ctcagggggc
     2101 agtatggaca agaagtacag catcggcctg gccatcggta ccaacagcgt gggctgggcc
     2161 gtgatcaccg acgagtacaa ggtgcccagc aagaagttca aggtgctggg caacaccgac
     2221 cgccacagca tcaagaagaa cctgatcggc gccctgctgt tcgacagcgg cgagaccgcc
     2281 gaggccaccc gcctgaagcg caccgcccgc cgccgctaca cccgccgcaa gaaccgcatc
     2341 tgctacctgc aggagatctt cagcaacgag atggccaagg tggacgacag cttcttccac
     2401 cgcctggagg agagcttcct ggtggaggag gacaagaagc acgagcgcca ccccatcttc
     2461 ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgcgc
     2521 aagaagctgg tggacagcac cgacaaggcc gacctgcgcc tgatctacct ggccctggcc
     2581 cacatgatca agttccgcgg ccacttcctg atcgagggcg acctgaaccc cgacaacagc
     2641 gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt cgaggagaac
     2701 cccatcaacg ccagcggcgt ggacgccaag gccatcctga gcgcccgcct gagcaagagc
     2761 cgccgcctgg agaacctgat cgcccagctg cccggcgaga agaagaacgg cctgttcggc
     2821 aacctgatcg ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc
     2881 gaggacgcca agctgcagct gagcaaggac acctacgacg acgacctgga caacctgctg
     2941 gcccagatcg gcgaccagta cgccgacctg ttcctggccg ccaagaacct gagcgacgcc
     3001 atcctgctga gcgacatcct gcgcgtgaac accgagatca ccaaggcccc cctgagcgcc
     3061 agcatgatca agcgctacga cgagcaccac caggacctga ccctgctgaa ggccctggtg
     3121 cgccagcagc tgcccgagaa gtacaaggag atcttcttcg accagagcaa gaacggctac
     3181 gccggctaca tcgacggcgg cgccagccag gaggagttct acaagttcat caagcccatc
     3241 ctggagaaga tggacggcac cgaggagctg ctggtgaagc tgaaccgcga ggacctgctg
     3301 cgcaagcagc gcaccttcga caacggcagc atcccccacc agatccacct gggcgagctg
     3361 cacgccatcc tgcgccgcca ggaggacttc taccccttcc tgaaggacaa ccgcgagaag
     3421 atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccccctggc ccgcggcaac
     3481 agccgcttcg cctggatgac ccgcaagagc gaggagacca tcaccccctg gaacttcgag
     3541 gaggtggtgg acaagggcgc cagcgcccag agcttcatcg agcgcatgac caacttcgac
     3601 aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc
     3661 gtgtacaacg agctgaccaa ggtgaagtac gtgaccgagg gcatgcgcaa gcccgccttc
     3721 ctgagcggcg agcagaagaa ggccatcgtg gacctgctgt tcaagaccaa ccgcaaggtg
     3781 accgtgaagc agctgaagga ggactacttc aagaagatcg agtgcttcga cagcgtggag
     3841 atcagcggcg tggaggaccg cttcaacgcc agcctgggca cctaccacga cctgctgaag
     3901 atcatcaagg acaaggactt cctggacaac gaggagaacg aggacatcct ggaggacatc
     3961 gtgctgaccc tgaccctgtt cgaggaccgc gagatgatcg aggagcgcct gaagacctac
     4021 gcccacctgt tcgacgacaa ggtgatgaag cagctgaagc gccgccgcta caccggctgg
     4081 ggccgcctga gccgcaagct tatcaacggc atccgcgaca agcagagcgg caagaccatc
     4141 ctggacttcc tgaagagcga cggcttcgcc aaccgcaact tcatgcagct gatccacgac
     4201 gacagcctga ccttcaagga ggacatccag aaggcccagg tgagcggcgt aagtatcaag
     4261 gttacaagac aggtttaagg agaccaatag aaactgggcg gaccgagcgg ccgcaggaac
     4321 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
     4381 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
     4441 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     4501 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     4561 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc
     4621 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4681 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4741 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4801 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4861 ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt
     4921 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     4981 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     5041 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     5101 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     5161 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     5221 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5281 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5341 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5401 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5461 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5521 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5581 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     5641 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5701 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5761 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5821 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5881 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     5941 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6001 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6061 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6121 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6181 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6241 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6301 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6361 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6421 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6481 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6541 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6601 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6661 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6721 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6781 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6841 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6901 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6961 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7021 cctggccttt tgctggcctt ttgctcacat gt
//
