LOCUS       sequence-369999-        8241 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AAV transgene plasmid for Cas7-11-R1045-GGGS-R1122, with U6 
            promoter-driven expression of Gluc crRNA guide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8241)
  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 8241)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pDF0428 huDisCas7-11-R1045-GGGS-R1122-U6-pro-Gluc
FEATURES             Location/Qualifiers
     source          1..8241
                     /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    7638..7686
                     /note="synthetic polyadenylation signal"
     promoter        7687..7935
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7687..7707
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7858..7877
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     repeat_region   8057..8186
                     /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 gagacccacg 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 tttccgaaac tgcaagggtg cctttctatg acaaaacgca gaaacatttt gacattcttg
     6121 atgagcaaga aattgctggt gagaaacctg ttcggatgtg ggtcaagcgt tttattaaac
     6181 gagggggtgg gtcaagagat agtagatacc aaaaggcctt tcaagagata cccgagaacg
     6241 acccagacgg atgggagtgt aaagagggct accttcatgt cgtcggcccc agcaaagtag
     6301 agtttagtga caagaaaggg gatgtgatca ataactttca aggaacactc ccatcagttc
     6361 ccaacgactg gaaaacaatt aggacgaacg attttaagaa taggaaaagg aagaatgaac
     6421 ctgtgttttg ttgtgaggat gataagggca attactatac catggctaaa tattgcgaaa
     6481 ccttcttctt cgatctgaag gagaacgagg aatacgagat ccccgagaaa gcaagaatca
     6541 aatacaaaga actgctcagg gtctataaca acaatcctca agcagtgccg gagagcgtat
     6601 ttcagtctag agttgcccgg gaaaacgtgg aaaagctgaa gtccggagat cttgtgtatt
     6661 tcaaacataa tgaaaagtac gtagaggaca tcgtcccagt gcggatttcc cgaactgtag
     6721 acgataggat gatcggcaaa cgtatgagcg ccgatctgcg gccgtgccat ggagattggg
     6781 tggaagatgg tgatctcagt gccttgaatg catatcccga gaaaagactc ctcttgcgcc
     6841 accccaaagg actctgccct gcttgccggc tctttggaac cggatcttac aagggcagag
     6901 tcaggtttgg attcgtgtca ctcgaaaacg atccggagtg gctgatccca ggcaagaatc
     6961 ccggcgatcc gtttcacggc gggccggtga tgctctcatt gttggaacgg cctcgcccga
     7021 cttggagtat accgggatcc gacaataagt ttaaagtgcc tggcagaaag ttttacgtcc
     7081 accaccacgc ctggaaaacc attaaggacg ggaaccatcc cacaacaggc aaagctattg
     7141 aacaaagccc taataaccgc actgtagaag ctctcgccgg cgggaattcc tttagcttcg
     7201 aaattgcctt tgagaacctg aaagaatggg agctgggttt gctcatccac agcctgcaac
     7261 tcgaaaaggg tctggcgcat aaacttggaa tggcaaagtc tatgggattt ggttcagttg
     7321 aaattgacgt cgaatcagtg cgcctgagaa aagattggaa gcaatggcgg aatggcaatt
     7381 ccgaaattcc caactggttg ggaaaaggat ttgctaaact gaaggaatgg ttccgggacg
     7441 agctcgattt tatagaaaat cttaagaaac ttctttggtt tcctgagggc gaccaagcac
     7501 cccgggtttg ctaccccatg ctgcgaaaga aggacgatcc taatgggaat agcggttacg
     7561 aagaactcaa agacggggaa ttcaagaaag aagatcggca gaagaagctg accacgccgt
     7621 ggacaccgtg ggcataaaat aaaatatctt tattttcatt acatctgtgt gttggttttt
     7681 tgtgtggagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
     7741 agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
     7801 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
     7861 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
     7921 aaaggacgaa acaccgttga tgtcacggaa ctgcagccag ctttccgggc attggcttcc
     7981 attttttttg ggcccgggat cttcctagag catggctacg tagataagta gcatggcggg
     8041 ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     8101 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     8161 gcctcagtga gcgagcgagc gcgcagctgc attaatgaat cggccaacgc gcggggagag
     8221 gcggtttgcg tattgggcgc t
//
