LOCUS       sequence-226982-        9426 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Confers constitutive expression of SpCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9426)
  AUTHORS   Shen MW, Arbab M, Hsu JY, Worstell D, Culbertson SJ, Krabbe O, Cassa
            CA, Liu DR, Gifford DK, Sherwood RI
  TITLE     Predictable and precise template-free CRISPR editing of pathogenic 
            variants.
  JOURNAL   Nature. 2018 Nov;563(7733):646-651. doi: 10.1038/s41586-018-0686-x. 
            Epub 2018 Nov 7.
  PUBMED    30405244
REFERENCE   2  (bases 1 to 9426)
  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"; doi: 
            "10.1038/s41586-018-0686-x"; journalName: "Nature"; date: "2018-11";
            volume: "563"; issue: "7733"; pages: "646-651"
COMMENT     Sequence Label: p2T-CAG-SpCas9-BlastR
FEATURES             Location/Qualifiers
     source          1..9426
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      10..19
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             19..84
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             91..111
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             136..4236
                     /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             136..4236
                     /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             4237..4284
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4312..4329)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4318..4525
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    4549..4670
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    4918..5039
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(5197..5595)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(5614..5661)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(5709..6038)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5723..5858
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5777..5796)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6023..6043
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     misc_feature    join(6053..6240,6241..6252)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(6309..6329)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6309..6327)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6348..6364)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6348..6364)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6361..6383)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6372..6388
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6396..6402,6403..6420,6421..6426))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6441..6462
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6579..6596)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6750..7338)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6830..6849)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7509..8300,8301..8369))
                     /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     8132..8151
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8370..8474)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8500..8955)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8637..8658)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8849..8868
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9082..9104
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9096..9113
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     9097..9113
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9123..9142
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        9123..9141
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(9172..9183,9184..9346)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     regulatory      9371..9380
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(9386..9419)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 tccaccggag ccaccatgga ctataaggac cacgacggag actacaagga tcatgatatt
       61 gattacaaag acgatgacga taagatggcc ccaaagaaga agcggaaggt cggtatccac
      121 ggagtcccag cagccgacaa gaagtacagc atcggcctgg acatcggcac caactctgtg
      181 ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc
      241 aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc
      301 gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag
      361 aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc
      421 ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac
      481 cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac
      541 cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg
      601 gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc
      661 gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc
      721 gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg
      781 agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc
      841 ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc
      901 gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac
      961 aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg
     1021 tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc
     1081 ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa
     1141 gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag
     1201 aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc
     1261 aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag
     1321 gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg
     1381 ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac
     1441 cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc
     1501 aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg
     1561 aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc
     1621 aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag
     1681 tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag
     1741 cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac
     1801 cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac
     1861 tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat
     1921 ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg
     1981 gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg
     2041 aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac
     2101 accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc
     2161 aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg
     2221 atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag
     2281 ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc
     2341 atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc
     2401 gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac
     2461 agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg
     2521 aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg
     2581 cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac
     2641 gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg
     2701 ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg
     2761 aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag
     2821 ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc
     2881 atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac
     2941 tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc
     3001 accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc
     3061 gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc
     3121 ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac
     3181 gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac
     3241 ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag
     3301 atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag
     3361 ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa
     3421 aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc
     3481 gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc
     3541 cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa
     3601 ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag
     3661 aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc
     3721 aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct
     3781 gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg
     3841 tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag
     3901 ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc
     3961 tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag
     4021 caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc
     4081 aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac
     4141 accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac
     4201 gagacacgga tcgacctgtc tcagctggga ggcgacaaaa ggccggcggc cacgaaaaag
     4261 gccggccagg caaaaaagaa aaagtaagaa ttcctagagc tcgctgatca gcctcgactg
     4321 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     4381 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     4441 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     4501 aagagaatag caggcatgct ggggagcggc cgcttcgagc agacatgata agatacattg
     4561 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     4621 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     4681 attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt taaagcaagt
     4741 aaaacctcta caaatgtggt aaaatcgcga taaggatccg ggctggcgta atagcggaga
     4801 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gcgataagga
     4861 tccgggctgg cgtaatagcg aagaggcccg caccgggccg ctcgaataca gacatgataa
     4921 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     4981 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagttg
     5041 gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca gccggcgtcc
     5101 cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc gaaatctcgt
     5161 agcacgtgtc agtcctgctc ctcggccacg aagtgcttag ccctcccaca cataaccaga
     5221 gggcagcaat tcacgaatcc caactgccgt cggctgtcca tcactgtcct tcactatggc
     5281 tttgatccca ggatgcagat cgagaagcac ctgtcggcac cgtccgcagg ggctcaagat
     5341 gcccctgttc tcatttccga tcgcgacgat acaagtcagg ttgccagctg ccgcagcagc
     5401 agcagtgccc agcaccacga gttctgcaca aggtccccca gtaaaatgat atacattgac
     5461 accagtgaag atgcggccgt cgctagagag agctgcgctg gcgacgctgt agtcttcaga
     5521 gatggggatg ctgttgattg tagccgttgc tctttcaatg agggtggatt cttcttgaga
     5581 caaaggcttg gccatggttt agttcctcac cttgtcgtat tatactatgc cgatatacta
     5641 tgccgatgat taattgtcaa cacgtgctga tcagatccga aaatggatat acaagctccc
     5701 gggagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaatagc
     5761 tcagaggcag aggcggcctc ggcctctgca taaataaaaa aaattagtca gccatggggc
     5821 ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag gggcgggact
     5881 atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
     5941 gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct
     6001 ggggagcctg gggactttcc acaccctaac tgacacacat tccacagaat tcaagtgatc
     6061 tccaaaaaat aagtactttt tgactgtaaa taaaattgta aggagtaaaa agtacttttt
     6121 tttctaaaaa aatgtaatta agtaaaagta aaagtattga tttttaattg tactcaagta
     6181 aagtaaaaat ccccaaaaat aatacttaag tacagtaatc aagtaaaatt actcaagtac
     6241 tttacacctc tggttcttga ccccctacct tcagcaagcc cagcagatcc gagctccagc
     6301 ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt
     6361 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     6421 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     6481 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6541 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6601 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6661 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6721 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6781 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6841 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6901 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6961 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     7021 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     7081 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     7141 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     7201 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     7261 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     7321 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     7381 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     7441 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7501 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     7561 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     7621 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7681 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7741 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7801 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     7861 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7921 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7981 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     8041 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     8101 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     8161 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     8221 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     8281 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     8341 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     8401 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     8461 ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt
     8521 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     8581 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     8641 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     8701 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     8761 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     8821 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     8881 cgcggggggg tgggggggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     8941 cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     9001 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     9061 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg
     9121 cgtaatacga ctcactatag ggcgaattgg gtaccggcat atggttcttg acagaggtgt
     9181 aaaaagtact caaaaatttt actcaagtga aagtacaagt acttagggaa aattttactc
     9241 aattaaaagt aaaagtatct ggctagaatc ttacttgagt aaaagtaaaa aagtactcca
     9301 ttaaaattgt acttgagtat taaggaagta aaagtaaaag caagaaagat cgatctcgaa
     9361 ggatctggag gccaccatgg tgtcgataac ttcgtatagc atacattata cgaagttatc
     9421 gataag
//
