LOCUS       sequence-196970-        9018 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  sgRNA/Cas9 expression plasmid with Cas9 VQR mutations (NGA PAM).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9018)
  AUTHORS   Lambrus BG, Moyer TC, Holland AJ
  TITLE     Applying the auxin-inducible degradation system for rapid protein 
            depletion in mammalian cells.
  JOURNAL   Methods Cell Biol. 2018;144:107-135. doi: 
            10.1016/bs.mcb.2018.03.004. Epub 2018 Apr 5.
  PUBMED    29804665
REFERENCE   2  (bases 1 to 9018)
  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: "Methods 
            Cell Biol."; date: "2018"; volume: "144:107-135. doi"; pages: " 
            10.1016/bs.mcb.2018.03.004. Epub 2018 Apr 5"
COMMENT     Sequence Label: px459 VQR
FEATURES             Location/Qualifiers
     source          1..9018
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          6..233
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      245..254
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             254..319
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             326..346
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(371..3769,3770..3772,3773..4369,4370..4372,4373..4375,
                     4376..4378,4379..4471)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/R1335Q/T1337R mutations"
                     /label=Cas9 VQR
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKQYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4472..4519
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4535..4588
                     /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             4589..5188
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4589..4608)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5085..5105
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5216..5233)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5222..5429
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5438..5578
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5438..5567
                     /label=AAV2 ITR
     rep_origin      5653..6108
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5740..5759)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5950..5971
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6125..6144)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6244..6266
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6304..6322)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6390..6494
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6495..6563,6564..7355)
                     /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(6713..6732)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7526..8114
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8015..8034
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8176..8424
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8176..8196
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8347..8366
                     /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 gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc
       61 gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt
      121 ctcctccggg ctgtaattag ctgagcaaga ggtaagggtt taagggatgg ttggttggtg
      181 gggtattaat gtttaattac ctggagcacc tgcctgaaat cacttttttt caggttggac
      241 cggtgccacc atggactata aggaccacga cggagactac aaggatcatg atattgatta
      301 caaagacgat gacgataaga tggccccaaa gaagaagcgg aaggtcggta tccacggagt
      361 cccagcagcc gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg
      421 ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac
      481 cgaccggcac agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac
      541 agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg
      601 gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt
      661 ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat
      721 cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct
      781 gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct
      841 ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa
      901 cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga
      961 aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa
     1021 gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt
     1081 cggaaacctg attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct
     1141 ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct
     1201 gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga
     1261 cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag
     1321 cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct
     1381 cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg
     1441 ctacgccggc tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc
     1501 catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct
     1561 gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga
     1621 gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga
     1681 aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg
     1741 aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt
     1801 cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt
     1861 cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt
     1921 caccgtgtat aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc
     1981 cttcctgagc ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa
     2041 agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt
     2101 ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct
     2161 gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga
     2221 tatagtgttg actcttaccc tctttgaaga tcgggaaatg attgaggaaa gactaaaaac
     2281 atacgctcac ctgttcgacg ataaggttat gaaacagtta aagaggcgtc gctatacggg
     2341 ctggggacga ttgtcgcgga aacttatcaa cgggataaga gacaagcaaa gtggtaaaac
     2401 tattctcgat tttctaaaga gcgacggctt cgccaatagg aactttatgc agctgatcca
     2461 tgatgactct ttaaccttca aagaggatat acaaaaggca caggtttccg gacaagggga
     2521 ctcattgcac gaacatattg cgaatcttgc tggttcgcca gccatcaaaa agggcatact
     2581 ccagacagtc aaagtagtgg atgagctagt taaggtcatg ggacgtcaca aaccggaaaa
     2641 cattgtaatc gagatggcac gcgaaaatca aacgactcag aaggggcaaa aaaacagtcg
     2701 agagcggatg aagagaatag aagagggtat taaagaactg ggcagccaga tcttaaagga
     2761 gcatcctgtg gaaaataccc aattgcagaa cgagaaactt tacctctatt acctacaaaa
     2821 tggaagggac atgtatgttg atcaggaact ggacataaac cgtttatctg attacgacgt
     2881 cgatcacatt gtaccccaat cctttttgaa ggacgattca atcgacaata aagtgcttac
     2941 acgctcggat aagaaccgag ggaaaagtga caatgttcca agcgaggaag tcgtaaagaa
     3001 aatgaagaac tattggcggc agctcctaaa tgcgaaactg ataacgcaaa gaaagttcga
     3061 taacttaact aaagctgaga ggggtggctt gtctgaactt gacaaggccg gatttattaa
     3121 acgtcagctc gtggaaaccc gccaaatcac aaagcatgtt gcacagatac tagattcccg
     3181 aatgaatacg aaatacgacg agaacgataa gctgattcgg gaagtcaaag taatcacttt
     3241 aaagtcaaaa ttggtgtcgg acttcagaaa ggattttcaa ttctataaag ttagggagat
     3301 aaataactac caccatgcgc acgacgctta tcttaatgcc gtcgtaggga ccgcactcat
     3361 taagaaatac ccgaagctag aaagtgagtt tgtgtatggt gattacaaag tttatgacgt
     3421 ccgtaagatg atcgcgaaaa gcgaacagga gataggcaag gctacagcca aatacttctt
     3481 ttattctaac attatgaatt tctttaagac ggaaatcact ctggcaaacg gagagatacg
     3541 caaacgacct ttaattgaaa ccaatgggga gacaggtgaa atcgtatggg ataagggccg
     3601 ggacttcgcg acggtgagaa aagttttgtc catgccccaa gtcaacatag taaagaaaac
     3661 tgaggtgcag accggagggt tttcaaagga atcgattctt ccaaaaagga atagtgataa
     3721 gctcatcgct cgtaaaaagg actgggaccc gaaaaagtac ggtggcttcg tgagccctac
     3781 agttgcctat tctgtcctag tagtggcaaa agttgagaag ggaaaatcca agaaactgaa
     3841 gtcagtcaaa gaattattgg ggataacgat tatggagcgc tcgtcttttg aaaagaaccc
     3901 catcgacttc cttgaggcga aaggttacaa ggaagtaaaa aaggatctca taattaaact
     3961 accaaagtat agtctgtttg agttagaaaa tggccgaaaa cggatgttgg ctagcgccgg
     4021 agagcttcaa aaggggaacg aactcgcact accgtctaaa tacgtgaatt tcctgtattt
     4081 agcgtcccat tacgagaagt tgaaaggttc acctgaagat aacgaacaga agcaactttt
     4141 tgttgagcag cacaaacatt atctcgacga aatcatagag caaatttcgg aattcagtaa
     4201 gagagtcatc ctagctgatg ccaatctgga caaagtatta agcgcataca acaagcacag
     4261 ggataaaccc atacgtgagc aggcggaaaa tattatccat ttgtttactc ttaccaacct
     4321 cggcgctcca gccgcattca agtattttga cacaacgata gatcgcaaac agtacagatc
     4381 taccaaggag gtgctagacg cgacactgat tcaccaatcc atcacgggat tatatgaaac
     4441 tcggatagat ttgtcacagc ttgggggtga caaaaggccg gcggccacga aaaaggccgg
     4501 ccaggcaaaa aagaaaaagg aattcggcag tggagagggc agaggaagtc tgctaacatg
     4561 cggtgacgtc gaggagaatc ctggcccaat gaccgagtac aagcccacgg tgcgcctcgc
     4621 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     4681 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     4741 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     4801 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     4861 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     4921 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggagtctc
     4981 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     5041 cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta
     5101 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     5161 gtgcatgacc cgcaagcccg gtgcctgaga attctaacta gagctcgctg atcagcctcg
     5221 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     5281 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     5341 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     5401 tgggaagaga atagcaggca tgctggggag cggccgcagg aacccctagt gatggagttg
     5461 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     5521 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     5581 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     5641 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     5701 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     5761 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     5821 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     5881 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     5941 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     6001 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     6061 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     6121 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     6181 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     6241 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     6301 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     6361 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     6421 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     6481 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     6541 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     6601 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     6661 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     6721 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     6781 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     6841 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     6901 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     6961 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     7021 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     7081 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     7141 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     7201 gtgagcgtgg aagccgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     7261 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     7321 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     7381 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     7441 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     7501 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     7561 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     7621 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     7681 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     7741 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7801 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     7861 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7921 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7981 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     8041 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     8101 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     8161 cttttgctca catgtgaggg cctatttccc atgattcctt catatttgca tatacgatac
     8221 aaggctgtta gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa
     8281 aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt
     8341 aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat
     8401 atcttgtgga aaggacgaaa caccgggtct tcgagaagac ctgttttaga 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 tgggggcggg gggggggg
//
