LOCUS       sequence-375888-        7392 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector expressing SaAID-2S and gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7392)
  AUTHORS   Li A, Mitsunobu H, Yoshioka S, Suzuki T, Kondo A, Nishida K
  TITLE     Cytosine base editing systems with minimized off-target effect and 
            molecular size.
  JOURNAL   Nat Commun. 2022 Aug 8;13(1):4531. doi: 10.1038/s41467-022-32157-8.
  PUBMED    35941130
REFERENCE   2  (bases 1 to 7392)
  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: "Nat 
            Commun."; date: "2022-08-8"; volume: "13(1):4531. doi"; pages: " 
            10.1038/s41467-022-32157-8"
COMMENT     Sequence Label: AAV_A2S-8
FEATURES             Location/Qualifiers
     source          1..7392
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      253..262
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             265..285
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3874..4122
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             4135..4182
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     polyA_signal    4216..4264
                     /note="synthetic polyadenylation signal"
     promoter        complement(4383..4631)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(4441..4460)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(4611..4631)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     repeat_region   4655..4795
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4655..4784
                     /label=AAV2 ITR
     rep_origin      4870..5325
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4957..4976)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5167..5188
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5342..5361)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5461..5483
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5521..5539)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5607..5711
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5712..5780,5781..6572)
                     /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(5930..5949)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6743..7331
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7232..7251
                     /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 gcggcctcta gactcgaggc gttgtactta tataaggggg tgggggcgcg
      181 ttcgtcctca gtcgcgatcg aacactcgag ccgagcagac gtgcctacgg accgctctgg
      241 ctaactaccg gtgccaccat ggccccaaag aagaagcgga aggtcggtat ccacggagtc
      301 ccagcagcct cacaccgctg ctacgtgctg tttgagttga agcgaagggg cgaaagaagg
      361 gcttgctttt ggggctatgc cgtcaacaag ccccaaagtg gcaccgagag aggaatacac
      421 gctgagatat tcagtatccg aaaggtggaa gagtatcttc gggataatcc tgggcagttt
      481 acgatcaact ggtattccag ctggagtcct tgcgctgatt gtgccgagaa aattctggaa
      541 tggtataatc aggaacttcg gggaaacggg cacacattga aaatcgaagc ctgcaagctg
      601 tactacgaga agaatgcccg gaaccagata ggactccaga atctgaggga caatggtgta
      661 ggcctgaacg tgggtggagg tggcagtgat gtcggcggag gctccaagcg gaactacatc
      721 ctgggcctgg ccatcggcat caccagcgtg ggctacggca tcatcgacta cgagacacgg
      781 gacgtgatcg atgccggcgt gcggctgttc aaagaggcca acgtggaaaa caacgagggc
      841 aggcggagca agagaggcgc cagaaggctg aagcggcgga ggcggcatag aatccagaga
      901 gtgaagaagc tgctgttcga ctacaacctg ctgaccgacc acagcgagct gagcggcatc
      961 aacccctacg aggccagagt gaagggcctg agccagaagc tgagcgagga agagttctct
     1021 gccgccctgc tgcacctggc caagagaaga ggcgtgcaca acgtgaacga ggtggaagag
     1081 gacaccggca acgagctgtc caccaaagag cagatcagcc ggaacagcaa ggccctggaa
     1141 gagaaatacg tggccgaact gcagctggaa cggctgaaga aagacggcga agtgcggggc
     1201 agcatcaaca gattcaagac cagcgactac gtgaaagaag ccaaacagct gctgaaggtg
     1261 cagaaggcct accaccagct ggaccagagc ttcatcgaca cctacatcga cctgctggaa
     1321 acccggcgga cctactatga gggacctggc gagggcagcc ccttcggctg gaaggacatc
     1381 aaagaatggt acgagatgct gatgggccac tgcacctact tccccgagga actgcggagc
     1441 gtgaagtacg cctacaacgc cgacctgtac aacgccctga acgacctgaa caatctcgtg
     1501 atcaccaggg acgagaacga gaagctggaa tattacgaga agttccagat catcgagaac
     1561 gtgttcaagc agaagaagaa gcccaccctg aagcagatcg ccaaagaaat cctcgtgaac
     1621 gaagaggata ttaagggcta cagagtgacc agcaccggca agcccgagtt caccaacctg
     1681 aaggtgtacc acgacatcaa ggacattacc gcccggaaag agattattga gaacgccgag
     1741 ctgctggatc agattgccaa gatcctgacc atctaccaga gcagcgagga catccaggaa
     1801 gaactgacca atctgaactc cgagctgacc caggaagaga tcgagcagat ctctaatctg
     1861 aagggctata ccggcaccca caacctgagc ctgaaggcca tcaacctgat cctggacgag
     1921 ctgtggcaca ccaacgacaa ccagatcgct atcttcaacc ggctgaagct ggtgcccaag
     1981 aaggtggacc tgtcccagca gaaagagatc cccaccaccc tggtggacga cttcatcctg
     2041 agccccgtcg tgaagagaag cttcatccag agcatcaaag tgatcaacgc catcatcaag
     2101 aagtacggcc tgcccaacga catcattatc gagctggccc gcgagaagaa ctccaaggac
     2161 gcccagaaaa tgatcaacga gatgcagaag cggaaccggc agaccaacga gcggatcgag
     2221 gaaatcatcc ggaccaccgg caaagagaac gccaagtacc tgatcgagaa gatcaagctg
     2281 cacgacatgc aggaaggcaa gtgcctgtac agcctggaag ccatccctct ggaagatctg
     2341 ctgaacaacc ccttcaacta tgaggtggac cacatcatcc ccagaagcgt gtccttcgac
     2401 aacagcttca acaacaaggt gctcgtgaag caggaagaaa acagcaagaa gggcaaccgg
     2461 accccattcc agtacctgag cagcagcgac agcaagatca gctacgaaac cttcaagaag
     2521 cacatcctga atctggccaa gggcaagggc agaatcagca agaccaagaa agagtatctg
     2581 ctggaagaac gggacatcaa caggttctcc gtgcagaaag acttcatcaa ccggaacctg
     2641 gtggatacca gatacgccac cagaggcctg atgaacctgc tgcggagcta cttcagagtg
     2701 aacaacctgg acgtgaaagt gaagtccatc aatggcggct tcaccagctt tctgcggcgg
     2761 aagtggaagt ttaagaaaga gcggaacaag gggtacaagc accacgccga ggacgccctg
     2821 atcattgcca acgccgattt catcttcaaa gagtggaaga aactggacaa ggccaaaaaa
     2881 gtgatggaaa accagatgtt cgaggaaaag caggccgaga gcatgcccga gatcgaaacc
     2941 gagcaggagt acaaagagat cttcatcacc ccccaccaga tcaagcacat taaggacttc
     3001 aaggactaca agtacagcca ccgggtggac aagaagccta atagagagct gattaacgac
     3061 accctgtact ccacccggaa ggacgacaag ggcaacaccc tgatcgtgaa caatctgaac
     3121 ggcctgtacg acaaggacaa tgacaagctg aaaaagctga tcaacaagag ccccgaaaag
     3181 ctgctgatgt accaccacga cccccagacc taccagaaac tgaagctgat tatggaacag
     3241 tacggcgacg agaagaatcc cctgtacaag tactacgagg aaaccgggaa ctacctgacc
     3301 aagtactcca aaaaggacaa cggccccgtg atcaagaaga ttaagtatta cggcaacaaa
     3361 ctgaacgccc atctggacat caccgacgac taccccaaca gcagaaacaa ggtcgtgaag
     3421 ctgtccctga agccctacag attcgacgtg tacctggaca atggcgtgta caagttcgtg
     3481 accgtgaaga atctggatgt gatcaaaaaa gaaaactact acgaagtgaa tagcaagtgc
     3541 tatgaggaag ctaagaagct gaagaagatc agcaaccagg ccgagtttat cgcctccttc
     3601 tacaacaacg atctgatcaa gatcaacggc gagctgtata gagtgatcgg cgtgaacaac
     3661 gacctgctga accggatcga agtgaacatg atcgacatca cctaccgcga gtacctggaa
     3721 aacatgaacg acaagaggcc ccccaggatc attaagacaa tcgcctccaa gacccagagc
     3781 attaagaagt acagcacaga cattctgggc aacctgtatg aagtgaaatc taagaagcac
     3841 cctcagatca tcaaaaaggg ctctggaggc agcaccaacc tgtctgacat catcgagaag
     3901 gagacaggca agcagctggt catccaggag agcatcctga tgctgcccga agaagtcgaa
     3961 gaagtgatcg gaaacaagcc tgagagcgat atcctggtcc ataccgccta cgacgagagt
     4021 accgacgaaa atgtgatgct gctgacatcc gacgccccag agtataagcc ctgggctctg
     4081 gtcatccagg attccaacgg agagaacaaa atcaaaatgc tgtctggcgg ctcaaaaagg
     4141 ccggcggcca cgaaaaaggc cggccaggca aaaaagaaaa agtaagaatt cctagagctc
     4201 gctgatcagc ctcgaaataa aagatcttta ttttcattag atctgtgtgt tggttttttg
     4261 tgtgggtacc aaaaaaatct cgccaacaag ttgacgagat aaacacggca ttttgccttg
     4321 ttttagtaga ttctgtaatt ttcattacag agtactaaaa ccgagacgca ttaatcgtct
     4381 cgggtgtttc gtcctttcca caagatatat aaagccaaga aatcgaaata ctttcaagtt
     4441 acggtaagca tatgatagtc cattttaaaa cataatttta aaactgcaaa ctacccaaga
     4501 aattattact ttctacgtca cgtattttgt actaatatct ttgtgtttac agtcaaatta
     4561 attctaatta tctctctaac agccttgtat cgtatatgca aatatgaagg aatcatggga
     4621 aataggccct cttcctgccc gaccttgcgg ccgcaggaac ccctagtgat ggagttggcc
     4681 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     4741 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     4801 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     4861 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4921 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     4981 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5041 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     5101 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5161 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     5221 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     5281 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     5341 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     5401 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     5461 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     5521 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     5581 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5641 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5701 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5761 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5821 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5881 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5941 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     6001 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     6061 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     6121 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     6181 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     6241 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     6301 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     6361 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     6421 agcgtggaag ccgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     6481 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     6541 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     6601 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6661 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6721 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6781 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6841 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6901 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6961 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     7021 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7081 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     7141 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7201 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7261 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7321 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7381 ttgctcacat gt
//
