LOCUS       pAC84-pCR8-dCas9        7023 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Gateway(R) entry vector with a cassette encoding HA epitope-tagged 
            catalytically inactive S. pyogenes dCas9.
ACCESSION   .
VERSION     .
KEYWORDS    pAC84-pCR8-dCas9
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7023)
  AUTHORS   Cheng AW, Wang H, Yang H, Shi L, Katz Y, Theunissen TW, Rangarajan 
            S, Shivalila CS, Dadon DB, Jaenisch R.
  TITLE     Multiplexed activation of endogenous genes by CRISPR-on, an 
            RNA-guided transcriptional activator system.
  JOURNAL   Cell Res. 2013;23:1163-71.
  PUBMED    23979020
REFERENCE   2  (bases 1 to 7023)
  AUTHORS   Jaenisch Lab / Addgene #48218
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7023)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Res.";
            date: "2013"; volume: "23"; pages: "1163-71"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(387..473)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /label=attL1
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             695..697
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             698..724
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="HA"
                     /translation="YPYDVPDYA"
     CDS             728..748
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T
                     antigen"
                     /note="SV40 NLS"
                     /translation="PKKKRKV"
     CDS             758..4858
                     /label=dCas9
                     /note="catalytically dead mutant of the Cas9 endonuclease
                     from the Streptococcus pyogenes Type II CRISPR/Cas system"
     CDS             4862..4882
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T
                     antigen"
                     /note="SV40 NLS"
                     /translation="PKKKRKV"
     protein_bind    complement(4902..5001)
                     /label=attL2
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(5019..5037)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5042..5058)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             5489..6277
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     rep_origin      6373..6961
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc gaattcgccc ttaccggtgc caccatgtac ccatacgatg ttccagatta
      721 cgcttcgccg aagaaaaagc gcaaggtcga agcgtccgac aagaagtaca gcatcggcct
      781 ggccatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag
      841 caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg
      901 agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag
      961 aagaagatac accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga
     1021 gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga
     1081 ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca
     1141 cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc
     1201 cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct
     1261 gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt
     1321 gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa
     1381 ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct
     1441 gcccggcgag aagaagaatg gcctgttcgg caacctgatt gccctgagcc tgggcctgac
     1501 ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga
     1561 cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct
     1621 gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa
     1681 caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca
     1741 ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga
     1801 gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca
     1861 ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact
     1921 gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag
     1981 catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt
     2041 ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc
     2101 ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag
     2161 cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca
     2221 gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc
     2281 caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata
     2341 cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt
     2401 ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt
     2461 caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc
     2521 ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa
     2581 tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag
     2641 agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa
     2701 gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg
     2761 catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc
     2821 caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca
     2881 gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg
     2941 cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa
     3001 agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac
     3061 cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa
     3121 agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga
     3181 gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga
     3241 catcaaccgg ctgtccgact acgatgtgga cgccatcgtg cctcagagct ttctgaagga
     3301 cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa
     3361 cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc
     3421 caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag
     3481 cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa
     3541 gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct
     3601 gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga
     3661 tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct
     3721 gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt
     3781 gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat
     3841 cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga
     3901 gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac
     3961 cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat
     4021 gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc
     4081 tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa
     4141 gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt
     4201 ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat
     4261 ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga
     4321 agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg
     4381 ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc
     4441 ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc
     4501 cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat
     4561 catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa
     4621 agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat
     4681 catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac
     4741 caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca
     4801 ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacag
     4861 ccccaagaag aagagaaagg tggaggccag ctaagaattc gacccagctt tcttgtacaa
     4921 agttggcatt ataaaaaata attgctcatc aatttgttgc aacgaacagg tcactatcag
     4981 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     5041 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     5101 acaagataaa aatatatcat catgcctcct ctagaccagc caggacagaa atgcctcgac
     5161 ttcgctgctg cccaaggttg ccgggtgacg cacaccgtgg aaacggatga aggcacgaac
     5221 ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg
     5281 caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat
     5341 ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca agcagcaagc
     5401 gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcaggg
     5461 cagtcgccct aaaacaaagt taaacatcat gagggaagcg gtgatcgccg aagtatcgac
     5521 tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt
     5581 acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata ttgatttgct
     5641 ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca acgacctttt
     5701 ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag tcaccattgt
     5761 tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc aatttggaga
     5821 atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg acattgatct
     5881 ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc cagcggcgga
     5941 ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg aaaccttaac
     6001 gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc ttacgttgtc
     6061 ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg
     6121 ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta gacaggctta
     6181 tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag aatttgtcca
     6241 ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa ccctcgagcc acccatgacc
     6301 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     6361 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6421 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6481 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     6541 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6601 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6661 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6721 ggagcgaacg acctacaccg aactgagata cctacagcgt gagcattgag aaagcgccac
     6781 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6841 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6901 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6961 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     7021 gtt
//
