LOCUS       sequence-369890-        7272 bp    DNA     circular SYN 14-APR-2025
DEFINITION  mammalian expression of cas7-11-r978-gggs-r1294.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7272)
  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 7272)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pDF0387 pcmv-hudiscas7-11-r978-gggs-r1294
FEATURES             Location/Qualifiers
     source          1..7272
                     /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             4796..4813
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4836..4853)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4842..5066
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5137..5153)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5137..5153)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5150..5172)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5161..5177
                     /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(5185..5191,5192..5209,5210..5215))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5230..5251
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5368..5385)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5539..6127)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5619..5638)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6298..7089,7090..7158))
                     /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     6921..6940
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7159..7263)
                     /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 ggggggtggg tcacgaactg tagacgatag gatgatcggc aaacgtatga
     3901 gcgccgatct gcggccgtgc catggagatt gggtggaaga tggtgatctc agtgccttga
     3961 atgcatatcc cgagaaaaga ctcctcttgc gccaccccaa aggactctgc cctgcttgcc
     4021 ggctctttgg aaccggatct tacaagggca gagtcaggtt tggattcgcg tcactcgaaa
     4081 acgatccgga gtggctgatc ccaggcaaga atcccggcga tccgtttcac ggcgggccgg
     4141 tgatgctctc attgttggaa cggcctcgcc cgacttggag tataccggga tccgacaata
     4201 agtttaaagt gcctggcaga aagttttacg tccaccacca cgcctggaaa accattaagg
     4261 acgggaacca tcccacaaca ggcaaagcta ttgaacaaag ccctaataac cgcactgtag
     4321 aagctctcgc cggcgggaat tcctttagct tcgaaattgc ctttgagaac ctgaaagaat
     4381 gggagctggg tttgctcatc cacagcctgc aactcgaaaa gggtctggcg cataaacttg
     4441 gaatggcaaa gtctatggga tttggttcag ttgaaattga cgtcgaatca gtgcgcctga
     4501 gaaaagattg gaagcaatgg cggaatggca attccgaaat tcccaactgg ttgggaaaag
     4561 gatttgctaa actgaaggaa tggttccggg acgagctcga ttttatagaa aatcttaaga
     4621 aacttctttg gtttcctgag ggcgaccaag caccccgggt ttgctacccc atgctgcgaa
     4681 agaaggacga tcctaatggg aatagcggtt acgaagaact caaagacggg gaattcaaga
     4741 aagaagatcg gcagaagaag ctgaccacgc cgtggacacc gtgggcataa ccggtcatca
     4801 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     4861 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     4921 tgtcctttcc taataaaatg agaaaattgc atcgcattgt ctgagtaggt gtcattctat
     4981 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     5041 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctcgat
     5101 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     5161 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagccta
     5221 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5281 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5341 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5401 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5461 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5521 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5581 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5641 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5701 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5761 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5821 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5881 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5941 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6001 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6061 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6121 atctcaagaa gatcctttga tcttttctac ggggtctgac actcagtgga acgaaaactc
     6181 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6241 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6301 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6361 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6421 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6481 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6541 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6601 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6661 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6721 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     6781 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     6841 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6901 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6961 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7021 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7081 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7141 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7201 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7261 gcgcacattt cc
//
