LOCUS       sequence-482883-        8947 bp    DNA     circular SYN 12-AUG-2025
DEFINITION  For user to clone their gRNA oligos to BbsI site. PITCh gRNA 
            expression cassette is already build in so there is no need to 
            perform Golden Gate assembly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8947)
  AUTHORS   Lin DW, Chung BP, Huang JW, Wang X, Huang L, Kaiser P
  TITLE     Microhomology based CRISPR tagging tools for protein tracking, 
            purification, and depletion.
  JOURNAL   J Biol Chem. 2019 May 28. pii: RA119.008422. doi: 
            10.1074/jbc.RA119.008422.
  PUBMED    31138654
REFERENCE   2  (bases 1 to 8947)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 12, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Chem. 2019 May 28. pii: RA119.008422. doi: 
            10.1074/jbc.RA119.008422."
COMMENT     Sequence Label: pX330-BbsI-PITCh
FEATURES             Location/Qualifiers
     source          1..8947
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        51..336
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        338..615
                     /label=chicken beta-actin promoter
     intron          616..843
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      855..864
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             864..929
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             936..956
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             981..5081
                     /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             981..5081
                     /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             5082..5129
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5157..5174)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5163..5370
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5379..5519
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5379..5508
                     /label=AAV2 ITR
     rep_origin      5594..6049
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5681..5700)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5891..5912
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6066..6085)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6185..6207
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6245..6263)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6331..6435
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6436..6504,6505..7296)
                     /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(6654..6673)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7467..8055
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7956..7975
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8117..8365
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        8384..8459
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        8557..8805
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        8826..8901
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 cgtttttagc gcgtgcgcca attctgcaga caaatggctc tagaggtacc cgttacataa
       61 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      121 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      181 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      241 ggtaaatggc ccgcctggca ttgtgcccag tacatgacct tatgggactt tcctacttgg
      301 cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc
      361 ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa
      421 ttattttgtg cagcgatggg ggcggggggg gggggggggc gcgcgccagg cggggcgggg
      481 cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc
      541 gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga
      601 agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc
      661 gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg
      721 gacggccctt ctcctccggg ctgtaattag ctgagcaaga ggtaagggtt taagggatgg
      781 ttggttggtg gggtattaat gtttaattac ctggagcacc tgcctgaaat cacttttttt
      841 caggttggac cggtgccacc atggactata aggaccacga cggagactac aaggatcatg
      901 atattgatta caaagacgat gacgataaga tggccccaaa gaagaagcgg aaggtcggta
      961 tccacggagt cccagcagcc gacaagaagt acagcatcgg cctggacatc ggcaccaact
     1021 ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc
     1081 tgggcaacac cgaccggcac agcatcaaga agaacctgat cggagccctg ctgttcgaca
     1141 gcggcgaaac agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac
     1201 ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg
     1261 acagcttctt ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc
     1321 ggcaccccat cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca
     1381 tctaccacct gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct
     1441 atctggccct ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga
     1501 accccgacaa cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc
     1561 tgttcgagga aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca
     1621 gactgagcaa gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga
     1681 atggcctgtt cggaaacctg attgccctga gcctgggcct gacccccaac ttcaagagca
     1741 acttcgacct ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc
     1801 tggacaacct gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga
     1861 acctgtccga cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg
     1921 cccccctgag cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc
     1981 tgaaagctct cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga
     2041 gcaagaacgg ctacgccggc tacattgacg gcggagccag ccaggaagag ttctacaagt
     2101 tcatcaagcc catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca
     2161 gagaggacct gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc
     2221 acctgggaga gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg
     2281 acaaccggga aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc
     2341 tggccagggg aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc
     2401 cctggaactt cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc atcgagcgga
     2461 tgaccaactt cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt
     2521 acgagtactt caccgtgtat aacgagctga ccaaagtgaa atacgtgacc gagggaatga
     2581 gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga
     2641 ccaaccggaa agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct
     2701 tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc
     2761 acgatctgct gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca
     2821 ttctggaaga tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac
     2881 ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga
     2941 gatacaccgg ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt
     3001 ccggcaagac aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc
     3061 agctgatcca cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg
     3121 gccagggcga tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga
     3181 agggcatcct gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca
     3241 agcccgagaa catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga
     3301 agaacagccg cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga
     3361 tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact
     3421 acctgcagaa tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg
     3481 actacgatgt ggaccatatc gtgcctcaga gctttctgaa ggacgactcc atcgacaaca
     3541 aggtgctgac cagaagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg
     3601 tcgtgaagaa gatgaagaac tactggcggc agctgctgaa cgccaagctg attacccaga
     3661 gaaagttcga caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg
     3721 gcttcatcaa gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc
     3781 tggactcccg gatgaacact aagtacgacg agaatgacaa gctgatccgg gaagtgaaag
     3841 tgatcaccct gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag
     3901 tgcgcgagat caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa
     3961 ccgccctgat caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg
     4021 tgtacgacgt gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca
     4081 agtacttctt ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg
     4141 gcgagatccg gaagcggcct ctgatcgaga caaacggcga aaccggggag atcgtgtggg
     4201 ataagggccg ggattttgcc accgtgcgga aagtgctgag catgccccaa gtgaatatcg
     4261 tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga
     4321 acagcgataa gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg
     4381 acagccccac cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca
     4441 agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg
     4501 agaagaatcc catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga
     4561 tcatcaagct gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg
     4621 cctctgccgg cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact
     4681 tcctgtacct ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga
     4741 aacagctgtt tgtggaacag cacaagcact acctggacga gatcatcgag cagatcagcg
     4801 agttctccaa gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca
     4861 acaagcaccg ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc
     4921 tgaccaatct gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga
     4981 ggtacaccag caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc
     5041 tgtacgagac acggatcgac ctgtctcagc tgggaggcga caaaaggccg gcggccacga
     5101 aaaaggccgg ccaggcaaaa aagaaaaagt aagaattcct agagctcgct gatcagcctc
     5161 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     5221 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     5281 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     5341 ttgggaagag aatagcaggc atgctgggga gcggccgcag gaacccctag tgatggagtt
     5401 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     5461 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     5521 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     5581 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5641 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     5701 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     5761 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     5821 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     5881 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     5941 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     6001 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     6061 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     6121 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     6181 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     6241 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     6301 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     6361 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     6421 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     6481 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     6541 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     6601 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     6661 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     6721 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     6781 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     6841 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     6901 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6961 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     7021 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     7081 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     7141 ggtgagcgtg gaagccgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     7201 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     7261 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     7321 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     7381 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     7441 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     7501 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     7561 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     7621 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     7681 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     7741 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     7801 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     7861 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     7921 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7981 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     8041 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     8101 ccttttgctc acatgtgagg gcctatttcc catgattcct tcatatttgc atatacgata
     8161 caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca
     8221 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
     8281 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
     8341 tatcttgtgg aaaggacgaa acaccgggtc ttcgagaaga cctgttttag agctagaaat
     8401 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
     8461 tttttgtttt agagctagaa atagcaagtt aaaataaggc tagtccgttt ttagcgcgtg
     8521 cgccaattct gcagacaaat ggctctagag gcatgtgagg gcctatttcc catgattcct
     8581 tcatatttgc atatacgata caaggctgtt agagagataa ttggaattaa tttgactgta
     8641 aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt
     8701 gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat
     8761 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccgcatc gtacgcgtac
     8821 gtgttgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa
     8881 aaagtggcac cgagtcggtg cttttttgtt ttagagctag aaatagcaag ttaaaataag
     8941 gctagtc
//
