LOCUS       sequence-452213-        8887 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SpCas9, the PITCh gRNA, and a sgRNA targeting the 
            N-terminus of MYO1C for knock-in..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8887)
  AUTHORS   Philip R, Sharma A, Matellan L, Erpf AC, Hsu WH, Tkach JM, Wyatt 
            HDM, Pelletier L
  TITLE     qTAG: an adaptable plasmid scaffold for CRISPR-based endogenous 
            tagging.
  JOURNAL   EMBO J. 2024 Dec 12. doi: 10.1038/s44318-024-00337-5.
  PUBMED    39668248
REFERENCE   2  (bases 1 to 8887)
  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: "EMBO J. 
            2024 Dec 12. doi: 10.1038/s44318-024-00337-5."
COMMENT     Sequence Label: pX330-PITCh-MYO1C
FEATURES             Location/Qualifiers
     source          1..8887
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      734..743
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             743..808
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             815..835
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             860..4960
                     /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             860..4960
                     /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             4961..5008
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5036..5053)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5042..5249
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5258..5398
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5258..5387
                     /label=AAV2 ITR
     rep_origin      5473..5928
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5560..5579)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5770..5791
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5945..5964)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6064..6086
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6124..6142)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6210..6314
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6315..6383,6384..7175)
                     /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(6533..6552)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7346..7934
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7835..7854
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7996..8244
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        8266..8341
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        8439..8687
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        8708..8783
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        join(8880..8887,1..278)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 aacttacagt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
       61 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      121 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      181 acggtaaatg gcccgcctgg cattgtgccc agtacatgac cttatgggac tttcctactt
      241 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct
      301 gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt
      361 aattattttg tgcagcgatg ggggcggggg gggggggggg ggcggcccct gggggggggg
      421 gggcccgaag gttccttttt gggggggggg gggggcgggg gcggccttaa aaaaggcgag
      481 gcgcgcggcg ggcgggggcc gcggcgcgcc gcctccgccc cgtgccccgc tccgccgccg
      541 cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg
      601 acggcccttc tcctccgggc tgtaattagc tgagcaagag gtaagggttt aagggatggt
      661 tggttggtgg ggtattaatg tttaattacc tggagcacct gcctgaaatc actttttttc
      721 aggttggacc ggtgccacca tggactataa ggaccacgac ggagactaca aggatcatga
      781 tattgattac aaagacgatg acgataagat ggccccaaag aagaagcgga aggtcggtat
      841 ccacggagtc ccagcagccg acaagaagta cagcatcggc ctggacatcg gcaccaactc
      901 tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct
      961 gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc tgttcgacag
     1021 cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg
     1081 gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga
     1141 cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg
     1201 gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat
     1261 ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta
     1321 tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa
     1381 ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct
     1441 gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag
     1501 actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa
     1561 tggcctgttc ggaaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa
     1621 cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct
     1681 ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa
     1741 cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc
     1801 ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct
     1861 gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag
     1921 caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt tctacaagtt
     1981 catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaacag
     2041 agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca
     2101 cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat tcctgaagga
     2161 caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg tgggccctct
     2221 ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc
     2281 ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca tcgagcggat
     2341 gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta
     2401 cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg agggaatgag
     2461 aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc tgttcaagac
     2521 caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt
     2581 cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca
     2641 cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa acgaggacat
     2701 tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga tcgaggaacg
     2761 gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga agcggcggag
     2821 atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg acaagcagtc
     2881 cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa acttcatgca
     2941 gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc aggtgtccgg
     3001 ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg ccattaagaa
     3061 gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa
     3121 gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga agggacagaa
     3181 gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg gcagccagat
     3241 cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt acctgtacta
     3301 cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc ggctgtccga
     3361 ctacgatgtg gaccatatcg tgcctcagag ctttctgaag gacgactcca tcgacaacaa
     3421 ggtgctgacc agaagcgaca agaaccgggg caagagcgac aacgtgccct ccgaagaggt
     3481 cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga ttacccagag
     3541 aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg ataaggccgg
     3601 cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg cacagatcct
     3661 ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg aagtgaaagt
     3721 gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt tttacaaagt
     3781 gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg tcgtgggaac
     3841 cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg actacaaggt
     3901 gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa
     3961 gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc tggccaacgg
     4021 cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga tcgtgtggga
     4081 taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt
     4141 gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc ccaagaggaa
     4201 cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg gcggcttcga
     4261 cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa
     4321 gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa gcagcttcga
     4381 gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa aggacctgat
     4441 catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga gaatgctggc
     4501 ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat atgtgaactt
     4561 cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata atgagcagaa
     4621 acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc agatcagcga
     4681 gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt ccgcctacaa
     4741 caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc tgtttaccct
     4801 gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag
     4861 gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct
     4921 gtacgagaca cggatcgacc tgtctcagct gggaggcgac aaaaggccgg cggccacgaa
     4981 aaaggccggc caggcaaaaa agaaaaagta agaattccta gagctcgctg atcagcctcg
     5041 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     5101 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     5161 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     5221 tgggaagaga atagcaggca tgctggggag cggccgcagg aacccctagt gatggagttg
     5281 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     5341 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     5401 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     5461 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     5521 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt
     5581 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     5641 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     5701 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     5761 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt
     5821 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt
     5881 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     5941 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     6001 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     6061 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     6121 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     6181 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     6241 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     6301 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     6361 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     6421 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     6481 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     6541 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     6601 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     6661 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     6721 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     6781 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     6841 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     6901 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     6961 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     7021 gtgagcgtgg aagccgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     7081 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     7141 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     7201 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     7261 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     7321 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     7381 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     7441 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     7501 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     7561 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7621 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     7681 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7741 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7801 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     7861 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7921 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     7981 cttttgctca catgtgaggg cctatttccc atgattcctt catatttgca tatacgatac
     8041 aaggctgtta gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa
     8101 aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt
     8161 aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat
     8221 atcttgtgga aaggacgaaa caccgggcgg tgagcgcact ctccagtttt agagctagaa
     8281 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
     8341 cttttttgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt ttttagcgcg
     8401 tgcgccaatt ctgcagacaa atggctctag aggcatgtga gggcctattt cccatgattc
     8461 cttcatattt gcatatacga tacaaggctg ttagagagat aattggaatt aatttgactg
     8521 taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt
     8581 ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt
     8641 atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccgca tcgtacgcgt
     8701 acgtgttgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg
     8761 aaaaagtggc accgagtcgg tgcttttttg ttttagagct agaaatagca agttaaaata
     8821 aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac aaatggctct agaggtaccc
     8881 gttacat
//
