LOCUS       sequence-378845-        4054 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-KamiCas9 vector expressing sgGFP and human HTT sgRNAs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4054)
  AUTHORS   Duarte F, Ramosaj M, Hasanovic E, Regio S, Sipion M, Rey M, Deglon N
  TITLE     Semi-automated workflows to quantify AAV transduction in various 
            brain areas and predict gene editing outcome for neurological 
            disorders.
  JOURNAL   Mol Ther Methods Clin Dev. 2023 Mar 27;29:254-270. doi: 
            10.1016/j.omtm.2023.03.013. eCollection 2023 Jun 8.
  PUBMED    37090478
REFERENCE   2  (bases 1 to 4054)
  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: "Mol Ther 
            Methods Clin Dev."; date: "2023-03-27"; volume: "29:254-270. doi"; 
            pages: " 10.1016/j.omtm.2023.03.013. eCollection 2023 Jun 8"
COMMENT     Sequence Label: pAAV2ss-U6-sgHTT1-7sk-sgCas9
FEATURES             Location/Qualifiers
     source          1..4054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     169..188
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        648..723
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(771..795)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(855..872)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    861..1068
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1123..1140)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1294..1882)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1374..1393)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2053..2844,2845..2913))
                     /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     2676..2695
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2914..3018)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3086..3104
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3142..3164)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3264..3283
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3350..3805)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3487..3508)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3699..3718
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3947..3963
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3948..3964
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3987..4011
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        join(4052..4054,1..246)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     join(4052..4054,1..18)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata
       61 attggaatta atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag
      121 taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg
      181 cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga
      241 aacaccgacc ctggaaaagc tgatgagttt cagagctatg ctggaaacag catagcaagt
      301 tgaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctttttttc
      361 atagccatgg gcggccgctc tagactgcag tatttagcat gccccaccca tctgcaaggc
      421 attctggata gtgtcaaaac agccggaaat caagtccgtt tatctcaaac tttagcattt
      481 tgggaataaa tgatatttgc tatgctggtt aaattagatt ttagttaaat ttcctgctga
      541 agctctagta cgataagtaa cttgacctaa gtgtaaagtt gagatttcct tcaggtttat
      601 atagcttgtg cgccgcctgg gtacctcgtc gccgaagaaa aagcgcagtt ttagagctag
      661 aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg
      721 tgcttttttt ggtacctcta gatggaagct tctcgagact agtaggcctg acccagcttt
      781 cttgtacaaa gtggtgataa ttcctgcagc ccgggggatc cactagttct agtcgactag
      841 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
      901 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
      961 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     1021 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggaga gatctgagga
     1081 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgactcgct
     1141 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     1201 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     1261 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     1321 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     1381 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     1441 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     1501 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     1561 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     1621 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     1681 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     1741 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     1801 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     1861 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     1921 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     1981 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2041 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2101 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     2161 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     2221 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     2281 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     2341 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     2401 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     2461 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     2521 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     2581 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     2641 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     2701 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     2761 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     2821 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     2881 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     2941 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3001 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     3061 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     3121 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     3181 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     3241 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     3301 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga aattgtaagc
     3361 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3421 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3481 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3541 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3601 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     3661 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     3721 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     3781 cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gctgcgcaac tgttgggaag
     3841 ggcgatcggt gcgggcctct tcgckakyrm gcsagsscgc agagagggag tggccaactc
     3901 catcactagg ggttcctcag atctgaattc ggtaccgggc cccccctcga ggtcgacggt
     3961 atcgataagc ttgatatcga attatcacaa gtttgtacaa aaaagcaggc ttcatcgatc
     4021 aatgcggccg ctgggatcca agcttccatg ggag
//
