LOCUS       sequence-343149-        3080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes HvsCas7-11 DR and golden gate site for spacer clonings in an
            AAV backbone with U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3080)
  AUTHORS   Ozcan A, Krajeski R, Ioannidi E, Lee B, Gardner A, Makarova KS, 
            Koonin EV, Abudayyeh OO, Gootenberg JS
  TITLE     Programmable RNA targeting with the single-protein CRISPR effector 
            Cas7-11.
  JOURNAL   Nature. 2021 Sep 6. pii: 10.1038/s41586-021-03886-5. doi: 
            10.1038/s41586-021-03886-5.
  PUBMED    34489594
REFERENCE   2  (bases 1 to 3080)
  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: "Nature. 
            2021 Sep 6. pii: 10.1038/s41586-021-03886-5. doi: 
            10.1038/s41586-021-03886-5."
COMMENT     Sequence Label: pDF0338 pAAV U6-HvsCas7-11 DR Rev-BpiI golden ...
FEATURES             Location/Qualifiers
     source          1..3080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(21..476)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(158..179)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     370..389
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   548..677
                     /label=AAV2 ITR
     promoter        complement(736..984)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(794..813)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(964..984)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     repeat_region   985..1114
                     /label=AAV2 ITR
     rep_origin      complement(1351..1939)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1431..1450)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2110..2901,2902..2970))
                     /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     2733..2752
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2971..3075)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
       61 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
      121 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
      181 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
      241 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
      301 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
      361 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
      421 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca
      481 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt
      541 acgccagctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc
      601 gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag tggccaactc
      661 catcactagg ggttcctagc ttaaaaaaaa ggtcttctcg aagacccaca tcaaccttat
      721 tatcctcctt caaacggtgt ttcgtccttt ccacaagata tataaagcca agaaatcgaa
      781 atactttcaa gttacggtaa gcatatgata gtccatttta aaacataatt ttaaaactgc
      841 aaactaccca agaaattatt actttctacg tcacgtattt tgtactaata tctttgtgtt
      901 tacagtcaaa ttaattccaa ttatctctct aacagccttg tatcgtatat gcaaatatga
      961 aggaatcatg ggaaataggc cctcaggaac ccctagtgat ggagttggcc actccctctc
     1021 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     1081 cccgggcggc ctcagtgagc gagcgagcgc gcagctgcat taatgaatcg gccaacgcgc
     1141 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     1201 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     1261 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     1321 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1381 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1441 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1501 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1561 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1621 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1681 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     1741 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     1801 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1861 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1921 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     1981 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2041 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2101 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2161 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     2221 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     2281 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     2341 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     2401 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     2461 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     2521 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     2581 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     2641 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     2701 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     2761 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     2821 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     2881 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     2941 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     3001 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3061 aataggggtt ccgcgcacat
//
