LOCUS       sequence-192102-        8830 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses DD-SpCas9 in mammalian cells and has a cloning site for an
            sgRNA. The FKBP12 L106P destabilization domain allows for inducible 
            stabilization of Cas9 through the addition of Shield-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8830)
  AUTHORS   Geisinger JM, Turan S, Hernandez S, Spector LP, Calos MP
  TITLE     In vivo blunt-end cloning through CRISPR/Cas9-facilitated 
            non-homologous end-joining.
  JOURNAL   Nucleic Acids Res. 2016 Jan 13. pii: gkv1542.
  PUBMED    26762978
REFERENCE   2  (bases 1 to 8830)
  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: "Nucleic 
            Acids Res. 2016 Jan 13. pii: gkv1542."
COMMENT     Sequence Label: pX330-BsaIx2-DD-Cas9
FEATURES             Location/Qualifiers
     source          1..8830
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          172..399
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      411..420
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(417..419,420..740)
                     /codon_start=1
                     /product="destabilization domain that can be stabilized by 
                     Shield1 in the ProteoTuner(TM) system"
                     /label=DD
                     /note="L106P mutant of FKBP12"
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKPE"
     CDS             747..812
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             819..839
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             864..4964
                     /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             864..4964
                     /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             4965..5012
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5040..5057)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5046..5253
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5262..5402
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5262..5391
                     /label=AAV2 ITR
     rep_origin      5477..5932
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5564..5583)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5774..5795
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5949..5968)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6068..6090
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6128..6146)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6214..6318
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6319..6387,6388..7179)
                     /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(6537..6556)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7350..7938
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7839..7858
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8000..8248
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8000..8020
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8171..8190
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8266..8341
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8438..8723
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        join(8725..8830,1..171)
                     /label=chicken beta-actin promoter
ORIGIN
        1 gggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg gggcggggcg
       61 aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg
      121 gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct
      181 gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc
      241 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
      301 aattagctga gcaagaggta agggtttaag ggatggttgg ttggtggggt attaatgttt
      361 aattacctgg agcacctgcc tgaaatcact ttttttcagg ttggaccggt gccaccatgg
      421 gagtgcaggt ggaaaccatc tccccaggag acgggcgcac cttccccaag cgcggccaga
      481 cctgtgtggt gcactacacc gggatgcttg aagatggaaa gaaagtcgat tcctcccggg
      541 acagaaacaa gccctttaag tttatgctag gcaagcagga ggtgatccga ggctgggaag
      601 aaggggttgc ccagatgagt gtgggtcaga gagccaaact gactatatct ccagattatg
      661 cctatggtgc cactgggcac ccaggcatca tcccaccaca tgccactctc gtcttcgatg
      721 tggagcttct aaaaccggaa gaattcgact ataaggacca cgacggagac tacaaggatc
      781 atgatattga ttacaaagac gatgacgata agatggcccc aaagaagaag cggaaggtcg
      841 gtatccacgg agtcccagca gccgacaaga agtacagcat cggcctggac atcggcacca
      901 actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg
      961 tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc ctgctgttcg
     1021 acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca
     1081 gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg
     1141 acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg
     1201 agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca
     1261 ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga
     1321 tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc
     1381 tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc
     1441 agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg
     1501 ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga
     1561 agaatggcct gttcggaaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga
     1621 gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg
     1681 acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca
     1741 agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca
     1801 aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc
     1861 tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc
     1921 agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa gagttctaca
     1981 agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga
     2041 acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga
     2101 tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga
     2161 aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc
     2221 ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca
     2281 ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc ttcatcgagc
     2341 ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc
     2401 tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg accgagggaa
     2461 tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac ctgctgttca
     2521 agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt
     2581 gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat
     2641 accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg
     2701 acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg
     2761 aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc
     2821 ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc
     2881 agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca
     2941 tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt
     3001 ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta
     3061 agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc
     3121 acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac
     3181 agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc
     3241 agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt
     3301 actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt
     3361 ccgactacga tgtggaccat atcgtgcctc agagctttct gaaggacgac tccatcgaca
     3421 acaaggtgct gaccagaagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag
     3481 aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag ctgattaccc
     3541 agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg
     3601 ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga
     3661 tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc cgggaagtga
     3721 aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca
     3781 aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg
     3841 gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca
     3901 aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg
     3961 ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca
     4021 acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt
     4081 gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc caagtgaata
     4141 tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga
     4201 ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct
     4261 tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt
     4321 ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct
     4381 tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc
     4441 tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc
     4501 tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga
     4561 acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc
     4621 agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc gagcagatca
     4681 gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg ctgtccgcct
     4741 acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc cacctgttta
     4801 ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga
     4861 agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg
     4921 gcctgtacga gacacggatc gacctgtctc agctgggagg cgacaaaagg ccggcggcca
     4981 cgaaaaaggc cggccaggca aaaaagaaaa agtaagaatt cctagagctc gctgatcagc
     5041 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     5101 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     5161 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     5221 ggattgggaa gagaatagca ggcatgctgg ggagcggccg caggaacccc tagtgatgga
     5281 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     5341 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     5401 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     5461 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     5521 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
     5581 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     5641 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     5701 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     5761 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
     5821 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
     5881 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     5941 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     6001 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     6061 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     6121 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     6181 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     6241 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     6301 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     6361 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     6421 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     6481 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     6541 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     6601 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6661 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6721 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6781 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6841 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     6901 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6961 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     7021 gccggtgagc gtggaagccg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     7081 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     7141 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     7201 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     7261 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     7321 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     7381 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7441 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     7501 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7561 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7621 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7681 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7741 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7801 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7861 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7921 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7981 tggccttttg ctcacatgtg agggcctatt tcccatgatt ccttcatatt tgcatatacg
     8041 atacaaggct gttagagaga taattggaat taatttgact gtaaacacaa agatattagt
     8101 acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg
     8161 ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt
     8221 atatatcttg tggaaaggac gaaacaccgg gagaccgagg tctctgtttt agagctagaa
     8281 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
     8341 cttttttgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt ttttagcgcg
     8401 tgcgccaatt ctgcagacaa atggctctag aggtacccgt tacataactt acggtaaatg
     8461 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaatagta acgccaatag
     8521 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     8581 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     8641 cctggcattg tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     8701 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
     8761 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
     8821 cgatgggggc
//
