LOCUS       sequence-344933-        9406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes human codon optimized catalytically dead 
            DisCas7-11-NES-huADAR2 fusion protein alone for expression in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9406)
  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 9406)
  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: pDF0237 pCMV - dead huDisCas7-11 - NES - huADAR2
FEATURES             Location/Qualifiers
     source          1..9406
                     /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             5733..5759
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             6930..6947
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6970..6987)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6976..7200
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7271..7287)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7271..7287)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7284..7306)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7295..7311
                     /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(7319..7325,7326..7343,7344..7349))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7364..7385
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7502..7519)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7673..8261)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7753..7772)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8432..9223,9224..9292))
                     /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     9055..9074
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9293..9397)
                     /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 cgcactgcaa 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 ggtacagcgg
     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 ggaagcctgc agctgcctcc acttgaaaga ctgacactgg
     5761 gatcccagct gcatttaccg caggttttag ctgacgctgt ctcacgcctg gtcctgggta
     5821 agtttggtga cctgaccgac aacttctcct cccctcacgc tcgcagaaaa gtgctggctg
     5881 gagtcgtcat gacaacaggc acagatgtta aagatgccaa ggtgataagt gtttctacag
     5941 gaacaaaatg tattaatggt gaatacatga gtgatcgtgg ccttgcatta aatgactgcc
     6001 atgcagaaat aatatctcgg agatccttgc tcagatttct ttatacacaa cttgagcttt
     6061 acttaaataa caaagatgat caaaaaagat ccatctttca gaaatcagag cgaggggggt
     6121 ttaggctgaa ggagaatgtc cagtttcatc tgtacatcag cacctctccc tgtggagatg
     6181 ccagaatctt ctcaccacat gagccaatcc tggaagaacc agcagataga cacccaaatc
     6241 gtaaagcaag aggacagcta cggaccaaaa tagagtctgg tcaggggacg attccagtgc
     6301 gctccaatgc gagcatccaa acgtgggacg gggtgctgca aggggagcgg ctgctcacca
     6361 tgtcctgcag tgacaagatt gcacgctgga acgtggtggg catccaggga tcactgctca
     6421 gcattttcgt ggagcccatt tacttctcga gcatcatcct gggcagcctt taccacgggg
     6481 accacctttc cagggccatg taccagcgga tctccaacat agaggacctg ccacctctct
     6541 acaccctcaa caagcctttg ctcagtggca tcagcaatgc agaagcacgg cagccaggga
     6601 aggcccccaa cttcagtgtc aactggacgg taggcgactc cgctattgag gtcatcaacg
     6661 ccacgactgg gaaggatgag ctgggccgcg cgtcccgcct gtgtaagcac gcgttgtact
     6721 gtcgctggat gcgtgtgcac ggcaaggttc cctcccactt actacgctcc aagattacca
     6781 agcccaacgt gtaccatgag tccaagctgg cggcaaagga gtaccaggcc gccaaggcgc
     6841 gtctgttcac agccttcatc aaggcggggc tgggggcctg ggtggagaag cccaccgagc
     6901 aggaccagtt ctcactcacg taaaccggtc atcatcacca tcaccattga gtttaaaccc
     6961 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     7021 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgagaaaa
     7081 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     7141 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     7201 cttctgaggc ggaaagaacc agctggggct cgataccgtc gacctctagc tagagcttgg
     7261 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     7321 acatacgagc cggaagcata aagtgtaaag cctagggtgc ctaatgagtg agctaactca
     7381 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     7441 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     7501 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     7561 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     7621 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     7681 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     7741 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     7801 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     7861 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     7921 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     7981 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     8041 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     8101 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     8161 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     8221 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     8281 ctacggggtc tgacactcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     8341 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     8401 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     8461 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     8521 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     8581 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     8641 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     8701 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     8761 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     8821 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     8881 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     8941 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     9001 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     9061 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     9121 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     9181 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     9241 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     9301 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     9361 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttcc
//
