LOCUS       sequence-451404-        8949 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SpCas9, the PITCh gRNA, and a sgRNA targeting the 
            C-terminus of MAPRE1 for knock-in..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8949)
  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 8949)
  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-MAPRE1
FEATURES             Location/Qualifiers
     source          1..8949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        271..346
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        444..692
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        713..788
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        885..1170
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1172..1448
                     /label=chicken beta-actin promoter
     intron          1449..1676
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1688..1697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1697..1762
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1769..1789
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1814..5914
                     /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             1814..5914
                     /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             5915..5962
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5990..6007)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5996..6203
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6212..6352
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6212..6341
                     /label=AAV2 ITR
     rep_origin      6427..6882
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6514..6533)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6724..6745
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6899..6918)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7018..7040
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7078..7096)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7164..7268
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7269..7337,7338..8129)
                     /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(7487..7506)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8300..8888
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8789..8808
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg ctttgtgata cctgatgaag gttttagagc tagaaatagc aagttaaaat
      301 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga
      361 gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca
      421 gacaaatggc tctagaggca tgtgagggcc tatttcccat gattccttca tatttgcata
      481 tacgatacaa ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat
      541 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
      601 tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg
      661 ctttatatat cttgtggaaa ggacgaaaca ccgcatcgta cgcgtacgtg ttgttttaga
      721 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
      781 gtcggtgctt ttttgtttta gagctagaaa tagcaagtta aaataaggct agtccgtttt
      841 tagcgcgtgc gccaattctg cagacaaatg gctctagagg tacccgttac ataacttacg
      901 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aatagtaacg
      961 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     1021 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     1081 tggcccgcct ggcattgtgc ccagtacatg accttatggg actttcctac ttggcagtac
     1141 atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact
     1201 ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt
     1261 tgtgcagcga tgggggcggg gggggggggg ggcgcgcgcc agggggggcg gggcggggcg
     1321 aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc
     1381 gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc
     1441 ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc
     1501 gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc
     1561 cttctcctcc gggctgtaat tagctgagca agaggtaagg gtttaaggga tggttggttg
     1621 gtggggtatt aatgtttaat tacctggagc acctgcctga aatcactttt tttcaggttg
     1681 gaccggtgcc accatggact ataaggacca cgacggagac tacaaggatc atgatattga
     1741 ttacaaagac gatgacgata agatggcccc aaagaagaag cggaaggtcg gtatccacgg
     1801 agtcccagca gccgacaaga agtacagcat cggcctggac atcggcacca actctgtggg
     1861 ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa
     1921 caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga
     1981 aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa
     2041 ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt
     2101 cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc
     2161 catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca
     2221 cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc
     2281 cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga
     2341 caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga
     2401 ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag
     2461 caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct
     2521 gttcggaaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga
     2581 cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa
     2641 cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc
     2701 cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct
     2761 gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc
     2821 tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa
     2881 cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa
     2941 gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga
     3001 cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg
     3061 agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg
     3121 ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag
     3181 gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa
     3241 cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa
     3301 cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta
     3361 cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc
     3421 cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg
     3481 gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc
     3541 cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct
     3601 gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga
     3661 agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa
     3721 aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac
     3781 cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa
     3841 gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat
     3901 ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg
     3961 cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat
     4021 cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga
     4081 gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag
     4141 ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa
     4201 agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca
     4261 gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga
     4321 tgtggaccat atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct
     4381 gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa
     4441 gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt
     4501 cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat
     4561 caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc
     4621 ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac
     4681 cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga
     4741 gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct
     4801 gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga
     4861 cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt
     4921 cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat
     4981 ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg
     5041 ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa
     5101 gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga
     5161 taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc
     5221 caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact
     5281 gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa
     5341 tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa
     5401 gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc
     5461 cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta
     5521 cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct
     5581 gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca gcgagttctc
     5641 caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct acaacaagca
     5701 ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa
     5761 tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac
     5821 cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga
     5881 gacacggatc gacctgtctc agctgggagg cgacaaaagg ccggcggcca cgaaaaaggc
     5941 cggccaggca aaaaagaaaa agtaagaatt cctagagctc gctgatcagc ctcgactgtg
     6001 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     6061 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     6121 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     6181 gagaatagca ggcatgctgg ggagcggccg caggaacccc tagtgatgga gttggccact
     6241 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     6301 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     6361 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     6421 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     6481 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     6541 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     6601 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     6661 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     6721 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     6781 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     6841 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     6901 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     6961 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     7021 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     7081 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     7141 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     7201 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     7261 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     7321 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     7381 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     7441 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     7501 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     7561 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     7621 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     7681 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     7741 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     7801 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     7861 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     7921 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     7981 gtggaagccg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     8041 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     8101 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     8161 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     8221 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     8281 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     8341 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     8401 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     8461 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     8521 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     8581 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     8641 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     8701 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     8761 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     8821 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     8881 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     8941 ctcacatgt
//
