LOCUS       sequence-194492-        7853 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Encodes dCas9(N) fused to AVPR2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7853)
  AUTHORS   Baeumler TA, Ahmed AA, Fulga TA
  TITLE     Engineering Synthetic Signaling Pathways with Programmable 
            dCas9-Based Chimeric Receptors.
  JOURNAL   Cell Rep. 2017 Sep 12;20(11):2639-2653. doi: 
            10.1016/j.celrep.2017.08.044.
  PUBMED    28903044
REFERENCE   2  (bases 1 to 7853)
  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: 
            "10.1016/j.celrep.2017.08"; journalName: "Cell Rep"; date: 
            "2017-09-12- 12"; volume: "20"; issue: "11"; pages: "2639-2653"
COMMENT     Sequence Label: pAVPR2_TCS(Q'L)_HA-dCas9(N)_P2A-Puro-WPRE
FEATURES             Location/Qualifiers
     source          1..7853
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1556..1582
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(1598..1621,1622..1624,1625..3199)
                     /codon_start=1
                     /product="N-terminal portion of catalytically inactive 
                     Streptococcus pyogenes dCas9 carrying the D10A mutation 
                     (Zetsche et al., 2015)"
                     /label=dCas9(N)
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMR"
     CDS             3230..3286
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(3293..3312)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3789..3809
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(3926..3946)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4344..4355)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4486..4503)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4492..4699
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4708..4848
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4708..4837
                     /label=AAV2 ITR
     rep_origin      4923..5378
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5010..5029)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5220..5241
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5395..5414)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5514..5536
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5574..5592)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5660..5764
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5765..5833,5834..6625)
                     /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     complement(5983..6002)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6796..7384
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7285..7304
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7455..7740
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga
       61 ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat
      121 tagctgagca agaggtaagg gtttaaggga tggttggttg gtggggtatt aatgtttaat
      181 tacctggagc acctgcctga aatcactttt tttcaggttg gaccggtgcg ccaccatgaa
      241 gacgatcatc gccctgagct acatcttctg cctggtattc gccgactaca aggacgatga
      301 tgacgccagc atcgatatgc tcatggcctc taccacaagc gctgtgcccg gacatccatc
      361 tcttccaagt cttccctcca acagctcaca ggagaggcct ctggataccc gggacccact
      421 tcttgcaaga gcggagctcg ccttgctctc catcgttttc gttgccgtcg cgttgagcaa
      481 tggtctggtg ctggccgccc tggcgcgacg aggccgccga ggccactggg cgcctattca
      541 tgtctttatc ggccatctct gtcttgcaga cctggccgtg gcgctgtttc aggtgctccc
      601 tcaactggct tggaaagcca ccgatagatt ccgggggcca gacgcactgt gtcgggccgt
      661 gaaatacctg cagatggtgg gaatgtatgc cagttcatat atgatacttg ctatgacctt
      721 ggacagacac agggctatct gtcgccccat gctggcgtat aggcacggtt ccggagccca
      781 ttggaacagg ccagttctgg ttgcgtgggc tttttccctg cttctgagct tgccccagtt
      841 gtttattttc gcacagagaa atgtggaggg cggatctggc gtgaccgatt gctgggcttg
      901 cttcgctgag ccatggggca gaaggaccta cgtgacgtgg attgctctca tggtgttcgt
      961 tgctcccacc ctggggatcg ccgcatgcca ggtcctgata ttcagagaaa tacacgcctc
     1021 cttggttcct gggccaagcg agcgccctgg tggaagaaga cggggtcgaa ggactggctc
     1081 tccaggcgaa ggcgcccacg tatcagctgc ggtagctaag accgtacgca tgacactggt
     1141 catcgtcgtc gtttacgttc tttgctgggc cccgtttttt ttggtgcagc tctgggcggc
     1201 ctgggacccc gaagcacccc tggaaggtgc tccattcgtt ctgctcatgc ttctcgcttc
     1261 actgaatagc tgtaccaatc cttggattta cgcctctttc tcctccagtg ttagctccga
     1321 gctgcggagt cttctctgtt gcgcccgggg caggaccccc ccaagcctgg gtccacagga
     1381 cgaatcctgc actaccgcct ccagttccct ggccaaagac acctcttcta tcgataccgg
     1441 tggacgcacc ccacccagcc tgggtcccca agatgagtcc tgcaccaccg ccagctcctc
     1501 cctggccaag gacacttcat cgaccggtga gaacctgtac ttccagctaa ctagttaccc
     1561 atacgacgta ccagattacg ctggaggttc tggatccgac aagaagtaca gcatcggcct
     1621 ggccatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag
     1681 caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg
     1741 agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag
     1801 aagaagatac accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga
     1861 gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga
     1921 ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca
     1981 cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc
     2041 cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct
     2101 gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt
     2161 gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa
     2221 ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct
     2281 gcccggcgag aagaagaatg gcctgttcgg caacctgatt gccctgagcc tgggcctgac
     2341 ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga
     2401 cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct
     2461 gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa
     2521 caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca
     2581 ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga
     2641 gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca
     2701 ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact
     2761 gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag
     2821 catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt
     2881 ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc
     2941 ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag
     3001 cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca
     3061 gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc
     3121 caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata
     3181 cgtgaccgag ggaatgagag gtggcggtgg ctcgggtacc ggatctggag ccaccaattt
     3241 ttctctgctg aagcaggctg gtgatgttga agaaaaccct ggacctgaat tcatgaccga
     3301 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
     3361 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
     3421 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
     3481 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
     3541 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
     3601 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
     3661 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
     3721 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
     3781 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
     3841 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag gggtaactaa gtaaggatcg
     3901 atccaagata tcgtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     3961 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     4021 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     4081 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     4141 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     4201 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     4261 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
     4321 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     4381 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     4441 ccctttgggc cgcctccccg ccaattccta gagctcgctg atcagcctcg actgtgcctt
     4501 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     4561 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     4621 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaga
     4681 atagcaggca tgctggggag cggccgcagg aacccctagt gatggagttg gccactccct
     4741 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     4801 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     4861 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     4921 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4981 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     5041 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     5101 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     5161 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5221 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     5281 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     5341 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     5401 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     5461 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     5521 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     5581 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     5641 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5701 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5761 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5821 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5881 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5941 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     6001 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     6061 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     6121 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     6181 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     6241 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     6301 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     6361 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     6421 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     6481 aagccgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     6541 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     6601 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     6661 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6721 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6781 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6841 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6901 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6961 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     7021 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     7081 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     7141 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     7201 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     7261 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     7321 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     7381 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     7441 catctagagg tacccgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     7501 gacccccgcc cattgacgtc aatagtaacg ccaataggga ctttccattg acgtcaatgg
     7561 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     7621 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattgtgc ccagtacatg
     7681 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     7741 gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca
     7801 attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg ggg
//
