LOCUS       sequence-344930-        8205 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Encodes human codon optimized DisCas7-11 protein alone for 
            expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8205)
  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 8205)
  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: "Nature. 
            2021 Sep 6. pii: 10.1038/s41586-021-03886-5. doi: 
            10.1038/s41586-021-03886-5."
COMMENT     Sequence Label: pDF0159 pCMV - huDisCas7-11 mammalian expression
FEATURES             Location/Qualifiers
     source          1..8205
                     /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             5729..5746
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5769..5786)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5775..5999
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6070..6086)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6070..6086)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6083..6105)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6094..6110
                     /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(6118..6124,6125..6142,6143..6148))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6163..6184
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6301..6318)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6472..7060)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6552..6571)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7231..8022,8023..8091))
                     /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     7854..7873
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8092..8196)
                     /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 gactgagcct cgttgatcct gctaaacacc
     4081 cccagaagaa acaagacaat aaatggaaaa gacgcaaaga aggcatcgcc acatttatag
     4141 agcaaaagaa tggctcttat tattttaacg tggtcaccaa taatggctgc acttctttcc
     4201 acttgtggca taaacctgac aattttgacc aggagaaact cgaaggcata cagaatggtg
     4261 agaaactgga ttgctgggta agagatagta gataccaaaa ggcctttcaa gagatacccg
     4321 agaacgaccc agacggatgg gagtgtaaag agggctacct tcatgtcgtc ggccccagca
     4381 aagtagagtt tagtgacaag aaaggggatg tgatcaataa ctttcaagga acactcccat
     4441 cagttcccaa cgactggaaa acaattagga cgaacgattt taagaatagg aaaaggaaga
     4501 atgaacctgt gttttgttgt gaggatgata agggcaatta ctataccatg gctaaatatt
     4561 gcgaaacctt cttcttcgat ctgaaggaga acgaggaata cgagatcccc gagaaagcaa
     4621 gaatcaaata caaagaactg ctcagggtct ataacaacaa tcctcaagca gtgccggaga
     4681 gcgtatttca gtctagagtt gcccgggaaa acgtggaaaa gctgaagtcc ggagatcttg
     4741 tgtatttcaa acataatgaa aagtacgtag aggacatcgt cccagtgcgg atttcccgaa
     4801 ctgtagacga taggatgatc ggcaaacgta tgagcgccga tctgcggccg tgccatggag
     4861 attgggtgga agatggtgat ctcagtgcct tgaatgcata tcccgagaaa agactcctct
     4921 tgcgccaccc caaaggactc tgccctgctt gccggctctt tggaaccgga tcttacaagg
     4981 gcagagtcag gtttggattc gcgtcactcg aaaacgatcc ggagtggctg atcccaggca
     5041 agaatcccgg cgatccgttt cacggcgggc cggtgatgct ctcattgttg gaacggcctc
     5101 gcccgacttg gagtataccg ggatccgaca ataagtttaa agtgcctggc agaaagtttt
     5161 acgtccacca ccacgcctgg aaaaccatta aggacgggaa ccatcccaca acaggcaaag
     5221 ctattgaaca aagccctaat aaccgcactg tagaagctct cgccggcggg aattccttta
     5281 gcttcgaaat tgcctttgag aacctgaaag aatgggagct gggtttgctc atccacagcc
     5341 tgcaactcga aaagggtctg gcgcataaac ttggaatggc aaagtctatg ggatttggtt
     5401 cagttgaaat tgacgtcgaa tcagtgcgcc tgagaaaaga ttggaagcaa tggcggaatg
     5461 gcaattccga aattcccaac tggttgggaa aaggatttgc taaactgaag gaatggttcc
     5521 gggacgagct cgattttata gaaaatctta agaaacttct ttggtttcct gagggcgacc
     5581 aagcaccccg ggtttgctac cccatgctgc gaaagaagga cgatcctaat gggaatagcg
     5641 gttacgaaga actcaaagac ggggaattca agaaagaaga tcggcagaag aagctgacca
     5701 cgccgtggac accgtgggca taaccggtca tcatcaccat caccattgag tttaaacccg
     5761 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     5821 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgagaaaat
     5881 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     5941 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     6001 ttctgaggcg gaaagaacca gctggggctc gataccgtcg acctctagct agagcttggc
     6061 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     6121 catacgagcc ggaagcataa agtgtaaagc ctagggtgcc taatgagtga gctaactcac
     6181 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     6241 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6301 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6361 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6421 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6481 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6541 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6601 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6661 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6721 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6781 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6841 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6901 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6961 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7021 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7081 tacggggtct gacactcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7141 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7201 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7261 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7321 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7381 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7441 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7501 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7561 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7621 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7681 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7741 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7801 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7861 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7921 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7981 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8041 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8101 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8161 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttcc
//
