LOCUS       sequence-175036-        9249 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  For cloning and expression of sgRNA together with expression of a 
            Cas9-Venus fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9249)
  AUTHORS   Yumlu S, Bashir S, Stumm J, Kuhn R
  TITLE     Efficient Gene Editing of Human Induced Pluripotent Stem Cells Using
            CRISPR/Cas9.
  JOURNAL   Methods Mol Biol. 2019;1961:137-151. doi: 
            10.1007/978-1-4939-9170-9_10.
  PUBMED    30912045
REFERENCE   2  (bases 1 to 9249)
  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: "Methods Mol
            Biol."; date: "2019"; volume: "1961:137-151. doi"; pages: " 
            10.1007/978-1-4939-9170-9_10"
COMMENT     Sequence Label: pU6-(BbsI)sgRNA_CAG-Cas9-venus-bpA
FEATURES             Location/Qualifiers
     source          1..9249
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      24..33
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             36..56
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             66..89
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             114..4214
                     /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             114..4214
                     /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             4224..4244
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4245..4247,4248..4250,4251..4964)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4551..4570)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4898..4919
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4986..5002)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4986..5002)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5016..5033)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5022..5246
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5295..5312)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5295..5311)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5304..5326)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(5452..5907)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5589..5610)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5801..5820
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5934..6038
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6039..6107,6108..6899)
                     /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(6257..6276)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7070..7658
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7559..7578
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7812..7829
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7946..7967
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7982..7987,7988..8005,8006..8012)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8020..8036
                     /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)."
     primer_bind     8025..8047
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8044..8060
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8044..8060
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8085..8333
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8085..8105
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8256..8275
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8352..8427
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8462..8841
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     9161..9183
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
ORIGIN
        1 taatacgact cactataggg gccgccacca tgggacctaa gaaaaagagg aaggtggcgg
       61 ccgctgacta caaggatgac gacgataaag gtatccacgg agtcccagca gccgacaaga
      121 agtacagcat cggcctggac atcggcacca actctgtggg ctgggccgtg atcaccgacg
      181 agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca
      241 agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag gccacccggc
      301 tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag
      361 agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt
      421 ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg
      481 acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg
      541 acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt
      601 tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc
      661 tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca
      721 gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa
      781 atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggcaac ctgattgccc
      841 tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac
      901 tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg
      961 accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg
     1021 acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga
     1081 gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc
     1141 ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacattg
     1201 acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg
     1261 acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga
     1321 ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc
     1381 ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc
     1441 tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct
     1501 ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca
     1561 agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca
     1621 acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tataacgagc
     1681 tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc
     1741 agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc
     1801 tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg
     1861 aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca
     1921 aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga
     1981 cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg
     2041 acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc
     2101 ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga
     2161 agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct
     2221 ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca
     2281 ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg
     2341 tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg
     2401 ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga
     2461 tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca
     2521 cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg
     2581 tggaccagga actggacatc aaccggctgt ccgactacga tgtggaccat atcgtgcctc
     2641 agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc gacaagaacc
     2701 ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc
     2761 ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg accaaggccg
     2821 agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa
     2881 cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg
     2941 acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt
     3001 ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg
     3061 cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc
     3121 tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca
     3181 agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga
     3241 actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg
     3301 agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt gccaccgtgc
     3361 ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg
     3421 gcttcagcaa agagtctatc ctgcccaaga ggaacagcga taagctgatc gccagaaaga
     3481 aggactggga ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc
     3541 tggtggtggc caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc
     3601 tggggatcac catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag
     3661 ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt
     3721 tcgagctgga aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa
     3781 acgaactggc cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga
     3841 agctgaaggg ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaagc
     3901 actacctgga cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg
     3961 acgctaatct ggacaaagtg ctgtccgcct acaacaagca ccgggataag cccatcagag
     4021 agcaggccga gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct
     4081 tcaagtactt tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg
     4141 acgccaccct gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc
     4201 agctgggagg cgacgaggga gctcccaaga aaaagcgcaa ggtaatggtg agcaagggcg
     4261 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     4321 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     4381 agctgatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctgg
     4441 gctacggcct gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     4501 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     4561 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     4621 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     4681 acaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact
     4741 tcaagatccg ccacaacatc gaggacggcg gcgtgcagct cgccgaccac taccagcaga
     4801 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt
     4861 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     4921 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtgataaaac gcgtaaatga
     4981 ttgcagatcc actagttcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca
     5041 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     5101 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     5161 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaga atagcaggca
     5221 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct gggctcgagg
     5281 ctagagcggc cgccactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     5341 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     5401 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gaaattgtaa
     5461 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     5521 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     5581 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     5641 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     5701 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     5761 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     5821 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     5881 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5941 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     6001 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     6061 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     6121 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     6181 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     6241 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     6301 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     6361 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     6421 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     6481 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     6541 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     6601 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     6661 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6721 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6781 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6841 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6901 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6961 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     7021 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     7081 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     7141 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     7201 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     7261 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     7321 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     7381 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     7441 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     7501 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     7561 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     7621 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     7681 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     7741 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     7801 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     7861 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     7921 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     7981 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     8041 acacaggaaa cagctatgac catgaggcgc gccccccctt caccgagggc ctatttccca
     8101 tgattccttc atatttgcat atacgataca aggctgttag agagataatt ggaattaatt
     8161 tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg
     8221 ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt
     8281 gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac accgggtctt
     8341 cgagaagacc tgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa
     8401 cttgaaaaag tggcaccgag tcggtgcttt ttttgatcca agggtggggc gcgccggatt
     8461 cgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     8521 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     8581 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     8641 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     8701 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     8761 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     8821 ttagtcatcg ctattaccat gggctcgggg gaggggcgcg gcggcccccg gagcgccggc
     8881 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     8941 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
     9001 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
     9061 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
     9121 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
     9181 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagatc
     9241 cttaattaa
//
