LOCUS       sequence-394536-        8205 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses inactive Cas7-11 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8205)
  AUTHORS   Strecker J, Demircioglu FE, Li D, Faure G, Wilkinson ME, Gootenberg 
            JS, Abudayyeh OO, Nishimasu H, Macrae RK, Zhang F
  TITLE     RNA-activated protein cleavage with a CRISPR-associated 
            endopeptidase.
  JOURNAL   Science. 2022 Nov 25;378(6622):874-881. doi: 
            10.1126/science.add7450. Epub 2022 Nov 3.
  PUBMED    36423276
REFERENCE   2  (bases 1 to 8205)
  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"; doi: 
            "10.1126/science.add7450"; journalName: "Science"; date: 
            "2022-11-25- 25"; volume: "378"; issue: "6622"; pages: "874-881"
COMMENT     Sequence Label: pCDNA3_dCas7-11
FEATURES             Location/Qualifiers
     source          1..8205
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(48..67)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        239..618
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..822
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     773..793
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     864..883
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        864..882
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5707..5724
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5747..5764)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5753..5977
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6048..6064)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6048..6064)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6061..6083)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6072..6088
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6096..6102,6103..6120,6121..6126))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6141..6162
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6279..6296)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6450..7038)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6530..6549)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7209..8000,8001..8069))
                     /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     7832..7851
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8070..8174)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatcgatc tcccgatccc ctagggtcta ctctcagtac aatctgctct
       61 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
      121 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
      181 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
      241 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      301 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      361 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      421 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      481 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      541 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      601 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      661 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      721 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      781 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag
      841 atccgctaga gatccgcggc cgctaatacg actcactata gggagagccg ccaccatgac
      901 gactactatg aagatttcaa ttgaattcct cgagccgttt cggatgacca aatggcagga
      961 aagcactagg cgaaacaaga acaacaagga atttgttcgc ggccaagctt ttgcaagatg
     1021 gcatagaaat aagaaggata acacaaaagg gcggccttac ataactggga cattgctgcg
     1081 gtccgcagta attagaagcg cggaaaacct gctgacattg agtgatggga aaattagcga
     1141 aaagacatgt tgtcctggga aatttgatac ggaagataaa gacaggttgc tgcaattgag
     1201 gcagcggtcc acccttcgtt ggaccgataa gaacccctgt ccagataacg ctgaaactta
     1261 ctgcccgttt tgcgaacttc tcgggagatc cggtaacgat ggaaagaagg ctgagaagaa
     1321 ggattggcgc tttcgtattc attttggcaa tctttccctt ccagggaagc ccgatttcga
     1381 cgggccaaaa gctataggca gccaacgggt acttaacagg gtcgatttca aatccggtaa
     1441 ggcccatgac ttctttaagg cttacgaagt cgaccatact aggtttcccc gcttcgaagg
     1501 ggagattacc atagataata aagtcagcgc tgaagccagg aaactgctct gtgattctct
     1561 caagtttacg gatcggctgt gtggagctct gtgcgtaatc aggttcgatg aatatacacc
     1621 agcagccgat agtgggaaac aaaccgaaaa tgtccaagca gaaccgaatg ctaatctcgc
     1681 tgaaaagacc gcagagcaaa ttattagcat tctggacgat aataagaaaa ccgaatatac
     1741 ccgcctgctt gctgatgcta ttcgttcact ccgccgctct agtaaacttg tcgctggctt
     1801 gccaaaggat catgacggga aggatgacca ttacctctgg gatgtcggca agaagaagaa
     1861 agacgaaaac tctgtcacta ttaggcaaat ccttacgacc tcagcagaca ccaaggaact
     1921 caagaatgcc ggaaaatgga gagaattctg cgagaagctg ggtgaagcgc tgtacctcaa
     1981 gagtaaagac atgagtggcg gcctgaaaat tactcggaga atactgggcg atgctgaatt
     2041 ccatggaaag cccgatcggc ttgaaaagag ccggtctgtg tcaatcggat ctgtgttgaa
     2101 agaaactgtg gtatgcggcg aactggtcgc taaaacgccc ttcttcttcg gagcgataga
     2161 cgaagatgca aaacaaaccg cactgcaagt actcctcact cccgataaca agtatagact
     2221 gccaagaagc gccgtgcgag gtatactccg tcgagatctt caaacctatt ttgatagccc
     2281 atgcaatgct gagttgggtg gacggccatg catgtgtaag acgtgtagga ttatgagagg
     2341 gatcacggtg atggatgcgc gcagtgagta caatgccccg ccagaaataa ggcatcgtac
     2401 ccgcattaat cccttcacag gcacggttgc ggaaggtgcc ctgtttaata tggaagtagc
     2461 ccccgagggg attgtctttc catttcaact ccggtaccgg ggctctgaag atgggctgcc
     2521 cgatgcactg aaaacggtgt tgaaatggtg ggctgagggg caggcattca tgagtggcgc
     2581 tgcctcaacc gggaagggcc gattccggat ggaaaatgct aaatatgaaa cgctggatct
     2641 gagcgacgag aatcaaagga atgactatct taagaattgg ggatggcgtg acgagaaggg
     2701 gctcgaggaa ctgaagaaac gactgaactc aggtctgcca gagcccggta attataggga
     2761 tccaaaatgg cacgagatta acgtttccat tgagatggca agccctttta ttaatggcga
     2821 cccaatccgc gcagccgtgg acaaacgtgg tacagcagtg gttaccttcg ttaagtataa
     2881 agctgaaggg gaagaggcga aacccgtatg tgcatacaag gccgaatctt ttagaggggt
     2941 gatcagaagt gccgtggcac gcattcatat ggaagatggc gtccctttga ctgagttgac
     3001 tcacagtgac tgtgaatgtc tcctgtgcca aatctttgga agtgagtatg aagccggcaa
     3061 aataaggttt gaagatctcg tattcgaaag tgacccggaa cctgtgacct tcgatcatgt
     3121 ggccatcgat agattcactg gcggtgcagc tgataagaag aaattcgatg attcccctct
     3181 gcccggtagc cctgcaagac cgctcatgtt gaaaggctcc ttctggatcc gcagggacgt
     3241 tctcgaggac gaagagtact gtaaggcact cggtaaggct cttgcagatg tgaataatgg
     3301 cctttatccc ctcggtggaa agagcgccat cggctacgga caggtcaaga gtctgggtat
     3361 aaagggagat gataagagga tttctcgcct catgaatcct gcctttgatg agacagatgt
     3421 agccgttcca gaaaagccca aaactgatgc cgaggttcgc atcgaggcag agaaagtata
     3481 ttacccacac tatttcgtcg aaccccataa gaaggtggaa cgcgaggaga aaccctgtgg
     3541 tcatcaaaag ttccacgagg ggcgactgac aggtaaaatt cggtgtaagc tcattaccaa
     3601 gacacccctc atcgtcccag atactagtaa tgacgatttc ttcagacctg cggataaaga
     3661 agctcggaag gaaaaggacg aatatcataa atcatatgct ttcttcagac ttcataaaca
     3721 aatcatgatt cccgggagcg aattgagagg aatggtgagt agtgtctacg aaactgtgac
     3781 aaattcttgc ttcaggatat ttgatgagac taaacggttg tcatggcgga tggatgctga
     3841 tcaccagaat gtgctgcaag actttctccc aggtcgagtg accgccgatg ggaaacatat
     3901 acaaaagttt tccgaaactg caagggtgcc tttctatgac aaaacgcaga aacattttga
     3961 cattcttgat gagcaagaaa ttgctggtga gaaacctgtt cggatgtggg tcaagcgttt
     4021 tattaaacga ctgagcctcg ttgatcctgc taaacacccc cagaagaaac aagacaataa
     4081 atggaaaaga cgcaaagaag gcatcgccac atttatagag caaaagaatg gctcttatta
     4141 ttttaacgtg gtcaccaata atggctgcac ttctttccac ttgtggcata aacctgacaa
     4201 ttttgaccag gagaaactcg aaggcataca gaatggtgag aaactggatt gctgggtaag
     4261 agatagtaga taccaaaagg cctttcaaga gatacccgag aacgacccag acggatggga
     4321 gtgtaaagag ggctaccttc atgtcgtcgg ccccagcaaa gtagagttta gtgacaagaa
     4381 aggggatgtg atcaataact ttcaaggaac actcccatca gttcccaacg actggaaaac
     4441 aattaggacg aacgatttta agaataggaa aaggaagaat gaacctgtgt tttgttgtga
     4501 ggatgataag ggcaattact ataccatggc taaatattgc gaaaccttct tcttcgatct
     4561 gaaggagaac gaggaatacg agatccccga gaaagcaaga atcaaataca aagaactgct
     4621 cagggtctat aacaacaatc ctcaagcagt gccggagagc gtatttcagt ctagagttgc
     4681 ccgggaaaac gtggaaaagc tgaagtccgg agatcttgtg tatttcaaac ataatgaaaa
     4741 gtacgtagag gacatcgtcc cagtgcggat ttcccgaact gtagacgata ggatgatcgg
     4801 caaacgtatg agcgccgatc tgcggccgtg ccatggagat tgggtggaag atggtgatct
     4861 cagtgccttg aatgcatatc ccgagaaaag actcctcttg cgccacccca aaggactctg
     4921 ccctgcttgc cggctctttg gaaccggatc ttacaagggc agagtcaggt ttggattcgc
     4981 gtcactcgaa aacgatccgg agtggctgat cccaggcaag aatcccggcg atccgtttca
     5041 cggcgggccg gtgatgctct cattgttgga acggcctcgc ccgacttgga gtataccggg
     5101 atccgacaat aagtttaaag tgcctggcag aaagttttac gtccaccacc acgcctggaa
     5161 aaccattaag gacgggaacc atcccacaac aggcaaagct attgaacaaa gccctaataa
     5221 ccgcactgta gaagctctcg ccggcgggaa ttcctttagc ttcgaaattg cctttgagaa
     5281 cctgaaagaa tgggagctgg gtttgctcat ccacagcctg caactcgaaa agggtctggc
     5341 gcataaactt ggaatggcaa agtctatggg atttggttca gttgaaattg acgtcgaatc
     5401 agtgcgcctg agaaaagatt ggaagcaatg gcggaatggc aattccgaaa ttcccaactg
     5461 gttgggaaaa ggatttgcta aactgaagga atggttccgg gacgagctcg attttataga
     5521 aaatcttaag aaacttcttt ggtttcctga gggcgaccaa gcaccccggg tttgctaccc
     5581 catgctgcga aagaaggacg atcctaatgg gaatagcggt tacgaagaac tcaaagacgg
     5641 ggaattcaag aaagaagatc ggcagaagaa gctgaccacg ccgtggacac cgtgggcata
     5701 accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct
     5761 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     5821 gccactccca ctgtcctttc ctaataaaat gagaaaattg catcgcattg tctgagtagg
     5881 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     5941 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     6001 tggggctcga taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     6061 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     6121 tgtaaagcct agggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     6181 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6241 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6301 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6361 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6421 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6481 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6541 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6601 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6661 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6721 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6781 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6841 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6901 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6961 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7021 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cactcagtgg
     7081 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7141 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7201 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     7261 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     7321 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7381 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7441 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7501 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     7561 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7621 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7681 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7741 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7801 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7861 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7921 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7981 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     8041 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     8101 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     8161 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
