LOCUS       sequence-372325-        7845 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV transgene plasmid for Cas7-11-S1006-GGGS-D1221, with U6 
            promoter-driven expression of non-targeting crRNA guide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7845)
  AUTHORS   Kato K, Zhou W, Okazaki S, Isayama Y, Nishizawa T, Gootenberg JS, 
            Abudayyeh OO, Nishimasu H
  TITLE     Structure and engineering of the type III-E CRISPR-Cas7-11 effector 
            complex.
  JOURNAL   Cell. 2022 May 25. pii: S0092-8674(22)00581-5. doi: 
            10.1016/j.cell.2022.05.003.
  PUBMED    35643083
REFERENCE   2  (bases 1 to 7845)
  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: "Cell. 2022 
            May 25. pii: S0092-8674(22)00581-5. doi: 
            10.1016/j.cell.2022.05.003."
COMMENT     Sequence Label: pDF0584 huDisCas7-11 S1006-GGGS-D1221 U6-NT guide
FEATURES             Location/Qualifiers
     source          1..7845
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1932..2387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2069..2090)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2281..2300
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2459..2588
                     /label=AAV2 ITR
     promoter        2829..3040
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     polyA_signal    7224..7272
                     /note="synthetic polyadenylation signal"
     promoter        7273..7521
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7273..7293
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7444..7463
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     repeat_region   7661..7790
                     /label=AAV2 ITR
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gcgacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag
     2641 gaagagtacc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     2701 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     2761 tgtatcagtg gtttgtctgg tcaactaggt cttgaaagga gtgggaattg gctccggtgc
     2821 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
     2881 caattgatcc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
     2941 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
     3001 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag ggccaccatg acgactacta
     3061 tgaagatttc aattgaattc ctcgagccgt ttcggatgac caaatggcag gaaagcacta
     3121 ggcgaaacaa gaacaacaag gaatttgttc gcggccaagc ttttgcaaga tggcatagaa
     3181 ataagaagga taacacaaaa gggcggcctt acataactgg gacattgctg cggtccgcag
     3241 taattagaag cgcggaaaac ctgctgacat tgagtgatgg gaaaattagc gaaaagacat
     3301 gttgtcctgg gaaatttgat acggaagata aagacaggtt gctgcaattg aggcagcggt
     3361 ccacccttcg ttggaccgat aagaacccct gtccagataa cgctgaaact tactgcccgt
     3421 tttgcgaact tctcgggaga tccggtaacg atggaaagaa ggctgagaag aaggattggc
     3481 gctttcgtat tcattttggc aatctttccc ttccagggaa gcccgatttc gacgggccaa
     3541 aagctatagg cagccaacgg gtacttaaca gggtcgattt caaatccggt aaggcccatg
     3601 acttctttaa ggcttacgaa gtcgaccata ctaggtttcc ccgcttcgaa ggggagatta
     3661 ccatagataa taaagtcagc gctgaagcca ggaaactgct ctgtgattct ctcaagttta
     3721 cggatcggct gtgtggagct ctgtgcgtaa tcaggttcga tgaatataca ccagcagccg
     3781 atagtgggaa acaaaccgaa aatgtccaag cagaaccgaa tgctaatctc gctgaaaaga
     3841 ccgcagagca aattattagc attctggacg ataataagaa aaccgaatat acccgcctgc
     3901 ttgctgatgc tattcgttca ctccgccgct ctagtaaact tgtcgctggc ttgccaaagg
     3961 atcatgacgg gaaggatgac cattacctct gggatatcgg caagaagaag aaagacgaaa
     4021 actctgtcac tattaggcaa atccttacga cctcagcaga caccaaggaa ctcaagaatg
     4081 ccggaaaatg gagagaattc tgcgagaagc tgggtgaagc gctgtacctc aagagtaaag
     4141 acatgagtgg cggcctgaaa attactcgga gaatactggg cgatgctgaa ttccatggaa
     4201 agcccgatcg gcttgaaaag agccggtctg tgtcaatcgg atctgtgttg aaagaaactg
     4261 tggtatgcgg cgaactggtc gctaaaacgc ccttcttctt cggagcgata gacgaagatg
     4321 caaaacaaac cgacctgcaa gtactcctca ctcccgataa caagtataga ctgccaagaa
     4381 gcgccgtgcg aggtatactc cgtcgagatc ttcaaaccta ttttgatagc ccatgcaatg
     4441 ctgagttggg tggacggcca tgcatgtgta agacgtgtag gattatgaga gggatcacgg
     4501 tgatggatgc gcgcagtgag tacaatgccc cgccagaaat aaggcatcgt acccgcatta
     4561 atcccttcac aggcacggtt gcggaaggtg ccctgtttaa tatggaagta gcccccgagg
     4621 ggattgtctt tccatttcaa ctccggtacc ggggctctga agatgggctg cccgatgcac
     4681 tgaaaacggt gttgaaatgg tgggctgagg ggcaggcatt catgagtggc gctgcctcaa
     4741 ccgggaaggg ccgattccgg atggaaaatg ctaaatatga aacgctggat ctgagcgacg
     4801 agaatcaaag gaatgactat cttaagaatt ggggatggcg tgacgagaag gggctcgagg
     4861 aactgaagaa acgactgaac tcaggtctgc cagagcccgg taattatagg gatccaaaat
     4921 ggcacgagat taacgtttcc attgagatgg caagcccttt tattaatggc gacccaatcc
     4981 gcgcagccgt ggacaaacgt ggtacagatg tggttacctt cgttaagtat aaagctgaag
     5041 gggaagaggc gaaacccgta tgtgcataca aggccgaatc ttttagaggg gtgatcagaa
     5101 gtgccgtggc acgcattcat atggaagatg gcgtcccttt gactgagttg actcacagtg
     5161 actgtgaatg tctcctgtgc caaatctttg gaagtgagta tgaagccggc aaaataaggt
     5221 ttgaagatct cgtattcgaa agtgacccgg aacctgtgac cttcgatcat gtggccatcg
     5281 atagattcac tggcggtgca gctgataaga agaaattcga tgattcccct ctgcccggta
     5341 gccctgcaag accgctcatg ttgaaaggct ccttctggat ccgcagggac gttctcgagg
     5401 acgaagagta ctgtaaggca ctcggtaagg ctcttgcaga tgtgaataat ggcctttatc
     5461 ccctcggtgg aaagagcgcc atcggctacg gacaggtcaa gagtctgggt ataaagggag
     5521 atgataagag gatttctcgc ctcatgaatc ctgcctttga tgagacagat gtagccgttc
     5581 cagaaaagcc caaaactgat gccgaggttc gcatcgaggc agagaaagta tattacccac
     5641 actatttcgt cgaaccccat aagaaggtgg aacgcgagga gaaaccctgt ggtcatcaaa
     5701 agttccacga ggggcgactg acaggtaaaa ttcggtgtaa gctcattacc aagacacccc
     5761 tcatcgtccc agatactagt aatgacgatt tcttcagacc tgcggataaa gaagctcgga
     5821 aggaaaagga cgaatatcat aaatcatatg ctttcttcag acttcataaa caaatcatga
     5881 ttcccgggag cgaattgaga ggaatggtga gtagtgtcta cgaaactgtg acaaattctt
     5941 gcttcaggat atttgatgag actaaacggt tgtcatggcg gatggatgct gatcaccaga
     6001 atgtgctgca agactttctc ccaggtcgag tgaccgccga tgggaaacat atacaaaagt
     6061 tttccggggg tgggtcagat ctgaaggaga acgaggaata cgagatcccc gagaaagcaa
     6121 gaatcaaata caaagaactg ctcagggtct ataacaacaa tcctcaagca gtgccggaga
     6181 gcgtatttca gtctagagtt gcccgggaaa acgtggaaaa gctgaagtcc ggagatcttg
     6241 tgtatttcaa acataatgaa aagtacgtag aggacatcgt cccagtgcgg atttcccgaa
     6301 ctgtagacga taggatgatc ggcaaacgta tgagcgccga tctgcggccg tgccatggag
     6361 attgggtgga agatggtgat ctcagtgcct tgaatgcata tcccgagaaa agactcctct
     6421 tgcgccaccc caaaggactc tgccctgctt gccggctctt tggaaccgga tcttacaagg
     6481 gcagagtcag gtttggattc gcgtcactcg aaaacgatcc ggagtggctg atcccaggca
     6541 agaatcccgg cgatccgttt cacggcgggc cggtgatgct ctcattgttg gaacggcctc
     6601 gcccgacttg gagtataccg ggatccgaca ataagtttaa agtgcctggc agaaagtttt
     6661 acgtccacca ccacgcctgg aaaaccatta aggacgggaa ccatcccaca acaggcaaag
     6721 ctattgaaca aagccctaat aaccgcactg tagaagctct cgccggcggg aattccttta
     6781 gcttcgaaat tgcctttgag aacctgaaag aatgggagct gggtttgctc atccacagcc
     6841 tgcaactcga aaagggtctg gcgcataaac ttggaatggc aaagtctatg ggatttggtt
     6901 cagttgaaat tgacgtcgaa tcagtgcgcc tgagaaaaga ttggaagcaa tggcggaatg
     6961 gcaattccga aattcccaac tggttgggaa aaggatttgc taaactgaag gaatggttcc
     7021 gggacgagct cgattttata gaaaatctta agaaacttct ttggtttcct gagggcgacc
     7081 aagcaccccg ggtttgctac cccatgctgc gaaagaagga cgatcctaat gggaatagcg
     7141 gttacgaaga actcaaagac ggggaattca agaaagaaga tcggcagaag aagctgacca
     7201 cgccgtggac accgtgggca taaaataaaa tatctttatt ttcattacat ctgtgtgttg
     7261 gttttttgtg tggagggcct atttcccatg attccttcat atttgcatat acgatacaag
     7321 gctgttagag agataattag aattaatttg actgtaaaca caaagatatt agtacaaaat
     7381 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa
     7441 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc
     7501 ttgtggaaag gacgaaacac cgttgatgtc acggaacggt aatgcctggc ttgtcgacgc
     7561 atagtctgtt tttttaagct tgggccgctc gaggggcccg ggatcttcct agagcatggc
     7621 tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag
     7681 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc
     7741 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcattaat
     7801 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgct
//
