LOCUS       sequence-369886-        7989 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of cas7-11-r1045-gggs-r1122.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7989)
  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 7989)
  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: pDF0385 pcmv-hudiscas7-11-r1045-gggs-r1122
FEATURES             Location/Qualifiers
     source          1..7989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(70..89)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        261..640
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        641..844
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     795..815
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     886..905
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        886..904
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5513..5530
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5553..5570)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5559..5783
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5854..5870)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5854..5870)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5867..5889)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5878..5894
                     /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(5902..5908,5909..5926,5927..5932))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5947..5968
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6085..6102)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6256..6844)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6336..6355)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7015..7806,7807..7875))
                     /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     7638..7657
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7876..7980)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccgaaaagtg ccacctgacg tcgacggatc gggagatcga tctcccgatc ccctagggtc
       61 tactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
      121 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
      181 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
      241 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
      301 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      361 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      421 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      481 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      541 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      601 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      661 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      721 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      781 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      841 agctggttta gtgaaccgtc agatccgcta gagatccgcg gccgctaata cgactcacta
      901 tagggagagc cgccaccatg acgactacta tgaagatttc aattgaattc ctcgagccgt
      961 ttcggatgac caaatggcag gaaagcacta ggcgaaacaa gaacaacaag gaatttgttc
     1021 gcggccaagc ttttgcaaga tggcatagaa ataagaagga taacacaaaa gggcggcctt
     1081 acataactgg gacattgctg cggtccgcag taattagaag cgcggaaaac ctgctgacat
     1141 tgagtgatgg gaaaattagc gaaaagacat gttgtcctgg gaaatttgat acggaagata
     1201 aagacaggtt gctgcaattg aggcagcggt ccacccttcg ttggaccgat aagaacccct
     1261 gtccagataa cgctgaaact tactgcccgt tttgcgaact tctcgggaga tccggtaacg
     1321 atggaaagaa ggctgagaag aaggattggc gctttcgtat tcattttggc aatctttccc
     1381 ttccagggaa gcccgatttc gacgggccaa aagctatagg cagccaacgg gtacttaaca
     1441 gggtcgattt caaatccggt aaggcccatg acttctttaa ggcttacgaa gtcgaccata
     1501 ctaggtttcc ccgcttcgaa ggggagatta ccatagataa taaagtcagc gctgaagcca
     1561 ggaaactgct ctgtgattct ctcaagttta cggatcggct gtgtggagct ctgtgcgtaa
     1621 tcaggttcga tgaatataca ccagcagccg atagtgggaa acaaaccgaa aatgtccaag
     1681 cagaaccgaa tgctaatctc gctgaaaaga ccgcagagca aattattagc attctggacg
     1741 ataataagaa aaccgaatat acccgcctgc ttgctgatgc tattcgttca ctccgccgct
     1801 ctagtaaact tgtcgctggc ttgccaaagg atcatgacgg gaaggatgac cattacctct
     1861 gggatatcgg caagaagaag aaagacgaaa actctgtcac tattaggcaa atccttacga
     1921 cctcagcaga caccaaggaa ctcaagaatg ccggaaaatg gagagaattc tgcgagaagc
     1981 tgggtgaagc gctgtacctc aagagtaaag acatgagtgg cggcctgaaa attactcgga
     2041 gaatactggg cgatgctgaa ttccatggaa agcccgatcg gcttgaaaag agccggtctg
     2101 tgtcaatcgg atctgtgttg aaagaaactg tggtatgcgg cgaactggtc gctaaaacgc
     2161 ccttcttctt cggagcgata gacgaagatg caaaacaaac cgacctgcaa gtactcctca
     2221 ctcccgataa caagtataga ctgccaagaa gcgccgtgcg aggtatactc cgtcgagatc
     2281 ttcaaaccta ttttgatagc ccatgcaatg ctgagttggg tggacggcca tgcatgtgta
     2341 agacgtgtag gattatgaga gggatcacgg tgatggatgc gcgcagtgag tacaatgccc
     2401 cgccagaaat aaggcatcgt acccgcatta atcccttcac aggcacggtt gcggaaggtg
     2461 ccctgtttaa tatggaagta gcccccgagg ggattgtctt tccatttcaa ctccggtacc
     2521 ggggctctga agatgggctg cccgatgcac tgaaaacggt gttgaaatgg tgggctgagg
     2581 ggcaggcatt catgagtggc gctgcctcaa ccgggaaggg ccgattccgg atggaaaatg
     2641 ctaaatatga aacgctggat ctgagcgacg agaatcaaag gaatgactat cttaagaatt
     2701 ggggatggcg tgacgagaag gggctcgagg aactgaagaa acgactgaac tcaggtctgc
     2761 cagagcccgg taattatagg gatccaaaat ggcacgagat taacgtttcc attgagatgg
     2821 caagcccttt tattaatggc gacccaatcc gcgcagccgt ggacaaacgt ggtacagatg
     2881 tggttacctt cgttaagtat aaagctgaag gggaagaggc gaaacccgta tgtgcataca
     2941 aggccgaatc ttttagaggg gtgatcagaa gtgccgtggc acgcattcat atggaagatg
     3001 gcgtcccttt gactgagttg actcacagtg actgtgaatg tctcctgtgc caaatctttg
     3061 gaagtgagta tgaagccggc aaaataaggt ttgaagatct cgtattcgaa agtgacccgg
     3121 aacctgtgac cttcgatcat gtggccatcg atagattcac tggcggtgca gctgataaga
     3181 agaaattcga tgattcccct ctgcccggta gccctgcaag accgctcatg ttgaaaggct
     3241 ccttctggat ccgcagggac gttctcgagg acgaagagta ctgtaaggca ctcggtaagg
     3301 ctcttgcaga tgtgaataat ggcctttatc ccctcggtgg aaagagcgcc atcggctacg
     3361 gacaggtcaa gagtctgggt ataaagggag atgataagag gatttctcgc ctcatgaatc
     3421 ctgcctttga tgagacagat gtagccgttc cagaaaagcc caaaactgat gccgaggttc
     3481 gcatcgaggc agagaaagta tattacccac actatttcgt cgaaccccat aagaaggtgg
     3541 aacgcgagga gaaaccctgt ggtcatcaaa agttccacga ggggcgactg acaggtaaaa
     3601 ttcggtgtaa gctcattacc aagacacccc tcatcgtccc agatactagt aatgacgatt
     3661 tcttcagacc tgcggataaa gaagctcgga aggaaaagga cgaatatcat aaatcatatg
     3721 ctttcttcag acttcataaa caaatcatga ttcccgggag cgaattgaga ggaatggtga
     3781 gtagtgtcta cgaaactgtg acaaattctt gcttcaggat atttgatgag actaaacggt
     3841 tgtcatggcg gatggatgct gatcaccaga atgtgctgca agactttctc ccaggtcgag
     3901 tgaccgccga tgggaaacat atacaaaagt tttccgaaac tgcaagggtg cctttctatg
     3961 acaaaacgca gaaacatttt gacattcttg atgagcaaga aattgctggt gagaaacctg
     4021 ttcggatgtg ggtcaagcgt tttattaaac gagggggtgg gtcaagagat agtagatacc
     4081 aaaaggcctt tcaagagata cccgagaacg acccagacgg atgggagtgt aaagagggct
     4141 accttcatgt cgtcggcccc agcaaagtag agtttagtga caagaaaggg gatgtgatca
     4201 ataactttca aggaacactc ccatcagttc ccaacgactg gaaaacaatt aggacgaacg
     4261 attttaagaa taggaaaagg aagaatgaac ctgtgttttg ttgtgaggat gataagggca
     4321 attactatac catggctaaa tattgcgaaa ccttcttctt cgatctgaag gagaacgagg
     4381 aatacgagat ccccgagaaa gcaagaatca aatacaaaga actgctcagg gtctataaca
     4441 acaatcctca agcagtgccg gagagcgtat ttcagtctag agttgcccgg gaaaacgtgg
     4501 aaaagctgaa gtccggagat cttgtgtatt tcaaacataa tgaaaagtac gtagaggaca
     4561 tcgtcccagt gcggatttcc cgaactgtag acgataggat gatcggcaaa cgtatgagcg
     4621 ccgatctgcg gccgtgccat ggagattggg tggaagatgg tgatctcagt gccttgaatg
     4681 catatcccga gaaaagactc ctcttgcgcc accccaaagg actctgccct gcttgccggc
     4741 tctttggaac cggatcttac aagggcagag tcaggtttgg attcgcgtca ctcgaaaacg
     4801 atccggagtg gctgatccca ggcaagaatc ccggcgatcc gtttcacggc gggccggtga
     4861 tgctctcatt gttggaacgg cctcgcccga cttggagtat accgggatcc gacaataagt
     4921 ttaaagtgcc tggcagaaag ttttacgtcc accaccacgc ctggaaaacc attaaggacg
     4981 ggaaccatcc cacaacaggc aaagctattg aacaaagccc taataaccgc actgtagaag
     5041 ctctcgccgg cgggaattcc tttagcttcg aaattgcctt tgagaacctg aaagaatggg
     5101 agctgggttt gctcatccac agcctgcaac tcgaaaaggg tctggcgcat aaacttggaa
     5161 tggcaaagtc tatgggattt ggttcagttg aaattgacgt cgaatcagtg cgcctgagaa
     5221 aagattggaa gcaatggcgg aatggcaatt ccgaaattcc caactggttg ggaaaaggat
     5281 ttgctaaact gaaggaatgg ttccgggacg agctcgattt tatagaaaat cttaagaaac
     5341 ttctttggtt tcctgagggc gaccaagcac cccgggtttg ctaccccatg ctgcgaaaga
     5401 aggacgatcc taatgggaat agcggttacg aagaactcaa agacggggaa ttcaagaaag
     5461 aagatcggca gaagaagctg accacgccgt ggacaccgtg ggcataaccg gtcatcatca
     5521 ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     5581 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     5641 cctttcctaa taaaatgaga aaattgcatc gcattgtctg agtaggtgtc attctattct
     5701 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     5761 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctcgatacc
     5821 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5881 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctaggg
     5941 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6001 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6061 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6121 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6181 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6241 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6301 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6361 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6421 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6481 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6541 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6601 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6661 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     6721 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6781 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6841 tcaagaagat cctttgatct tttctacggg gtctgacact cagtggaacg aaaactcacg
     6901 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6961 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7021 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7081 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7141 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7201 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7261 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7321 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7381 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7441 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7501 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7561 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7621 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7681 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7741 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7801 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7861 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7921 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7981 cacatttcc
//
