LOCUS       sequence-171518-        9009 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses sgRNA targeting human EGFR around codon T790.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9009)
  AUTHORS   Guernet A, Mungamuri SK, Cartier D, Sachidanandam R, Jayaprakash A, 
            Adriouch S, Vezain M, Charbonnier F, Rohkin G, Coutant S, Yao S, 
            Ainani H, Alexandre D, Tournier I, Boyer O, Aaronson SA, Anouar Y, 
            Grumolato L
  TITLE     CRISPR-Barcoding for Intratumor Genetic Heterogeneity Modeling and 
            Functional Analysis of Oncogenic Driver Mutations.
  JOURNAL   Mol Cell. 2016 Aug 4;63(3):526-38. doi: 
            10.1016/j.molcel.2016.06.017. Epub 2016 Jul 21.
  PUBMED    27453044
REFERENCE   2  (bases 1 to 9009)
  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.molcel.2016.06.017"; journalName: "Mol Cell"; date: 
            "2016-08-4- 4"; volume: "63"; issue: "3"; pages: "526-38"
COMMENT     Sequence Label: sgEGFR(A)
FEATURES             Location/Qualifiers
     source          1..9009
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          3..230
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      242..251
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             251..316
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             323..343
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             368..4468
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             368..4468
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4469..4516
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4532..4585
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4586..5185
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4586..4605)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     complement(5213..5230)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5219..5426
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5435..5575
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5435..5564
                     /label=AAV2 ITR
     rep_origin      5650..6105
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5737..5756)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5947..5968
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6122..6141)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6241..6263
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6301..6319)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6387..6491
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6492..6560,6561..7352)
                     /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(6710..6729)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7523..8111
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8012..8031
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8173..8421
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8173..8193
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8344..8363
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8443..8518
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8615..8900
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
       61 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      121 ctccgggctg taattagctg agcaagaggt aagggtttaa gggatggttg gttggtgggg
      181 tattaatgtt taattacctg gagcacctgc ctgaaatcac tttttttcag gttggaccgg
      241 tgccaccatg gactataagg accacgacgg agactacaag gatcatgata ttgattacaa
      301 agacgatgac gataagatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc
      361 agcagccgac aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc
      421 cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga
      481 ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc
      541 cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat
      601 ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca
      661 cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt
      721 cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag
      781 aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc
      841 ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag
      901 cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa
      961 ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag
     1021 cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg
     1081 aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc
     1141 cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct
     1201 ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc
     1261 catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc
     1321 ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt
     1381 gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta
     1441 cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat
     1501 cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct
     1561 gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct
     1621 gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa
     1681 gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa
     1741 cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga
     1801 ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga
     1861 taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac
     1921 cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt
     1981 cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt
     2041 gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga
     2101 aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa
     2161 aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat
     2221 cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta
     2281 tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg
     2341 gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat
     2401 cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga
     2461 cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag
     2521 cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca
     2581 gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat
     2641 cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga
     2701 gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca
     2761 ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg
     2821 gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga
     2881 ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag
     2941 aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat
     3001 gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa
     3061 tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag
     3121 acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat
     3181 gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa
     3241 gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa
     3301 caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa
     3361 aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg
     3421 gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta
     3481 cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa
     3541 gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga
     3601 ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga
     3661 ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct
     3721 gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt
     3781 ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag
     3841 tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat
     3901 cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc
     3961 taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga
     4021 actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc
     4081 cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt
     4141 ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag
     4201 agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga
     4261 taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg
     4321 agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac
     4381 caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg
     4441 gatcgacctg tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa aggccggcca
     4501 ggcaaaaaag aaaaaggaat tcggcagtgg agagggcaga ggaagtctgc taacatgcgg
     4561 tgacgtcgag gagaatcctg gcccaatgac cgagtacaag cccacggtgc gcctcgccac
     4621 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     4681 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     4741 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     4801 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     4861 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     4921 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     4981 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     5041 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cacaacctcc ccttctacga
     5101 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     5161 catgacccgc aagcccggtg cctgagaatt ctaactagag ctcgctgatc agcctcgact
     5221 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     5281 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     5341 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     5401 gaagagaata gcaggcatgc tggggagcgg ccgcaggaac ccctagtgat ggagttggcc
     5461 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     5521 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     5581 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     5641 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     5701 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     5761 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5821 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     5881 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5941 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     6001 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     6061 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     6121 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     6181 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     6241 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     6301 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     6361 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     6421 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     6481 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     6541 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     6601 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     6661 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     6721 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     6781 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     6841 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     6901 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     6961 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     7021 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     7081 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     7141 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     7201 agcgtggaag ccgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     7261 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     7321 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     7381 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     7441 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     7501 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     7561 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     7621 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     7681 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     7741 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     7801 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7861 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     7921 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7981 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     8041 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     8101 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     8161 ttgctcacat gtgagggcct atttcccatg attccttcat atttgcatat acgatacaag
     8221 gctgttagag agataattgg aattaatttg actgtaaaca caaagatatt agtacaaaat
     8281 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa
     8341 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc
     8401 ttgtggaaag gacgaaacac cgctgcgtga tgagctgcac gggttttaga gctagaaata
     8461 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
     8521 ttttgtttta gagctagaaa tagcaagtta aaataaggct agtccgtttt tagcgcgtgc
     8581 gccaattctg cagacaaatg gctctagagg tacccgttac ataacttacg gtaaatggcc
     8641 cgcctggctg accgcccaac gacccccgcc cattgacgtc aatagtaacg ccaataggga
     8701 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     8761 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     8821 ggcattgtgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     8881 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
     8941 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
     9001 tgggggcgg
//
