LOCUS       sequence-256160-       10520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pDEF-SpCas9, Empty Vector for CRISPRi, PbEF1a promoter, 1st 
            Generation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10520)
  AUTHORS   Walker MP, Lindner SE
  TITLE     Ribozyme-mediated, multiplex CRISPR gene editing and CRISPR 
            interference (CRISPRi) in rodent-infectious Plasmodium yoelii.
  JOURNAL   J Biol Chem. 2019 Jun 14;294(24):9555-9566. doi: 
            10.1074/jbc.RA118.007121. Epub 2019 May 1.
  PUBMED    31043479
REFERENCE   2  (bases 1 to 10520)
  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: 
            "10.1074/jbc.RA118.007121"; journalName: "J Biol Chem"; date: 
            "2019-06-14- 14"; volume: "294"; issue: "24"; pages: "9555-9566"
COMMENT     Sequence Label: pSL1045
FEATURES             Location/Qualifiers
     source          1..10520
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1658..1684
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1688..1708
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1718..1741,1742..1744,1745..4300,4301..4303,
                     4304..5818)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(1718..1741,1742..1744,1745..4231,4232..4234,
                     4235..5818)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5822..5842
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     5894..5914
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(6483..6502)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6602..6624
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6662..6680)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6748..6852
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6853..6921,6922..7713)
                     /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(7071..7090)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7884..8472
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8373..8392
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8626..8643
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8760..8781
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8796..8801,8802..8819,8820..8826)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8834..8850
                     /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     8839..8861
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8858..8874
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8858..8874
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ggtaccccgc ggtggcggcc gcagcttaat tcttttcgag ctctttatgc ttaagtttac
       61 aatttaatat tcatacttta agtatttttt gtagtatcct agatattgtg ctttaaatgc
      121 tcacccctca aagcaccagt aatattttca tccactgaaa taccattaaa ttttcaaaaa
      181 aatactatgc atataatgtt atacatataa acataaaacg ccatgtaaat caaaaaatat
      241 ataaaaatat gtataaaaat aaatatgcac taaatataag ctaattatgc ataaaaatta
      301 aagtgccctt tattaactag ctagtcgtaa ttatttatat ttctatgtta taaaaaaatc
      361 ctcatataat aatataatta atatatgtaa tgtttttttt attttataat tttaatataa
      421 aataatatgt aaattaattc aaaaaataaa tataattgtt gtgaaacaaa aaacgtaatt
      481 ttttcatttg ccttcaaaat ttaaatttat tttaatattt cctaaaatat atatactttg
      541 tgtataaata tataaaaata tatatttgct tataaataaa taaaaatttt ataaaaatgg
      601 ctagcaggcc taccggtgtt tttcttactt atatatttat accaattgat tgtatttata
      661 actgtaaaaa tgtgtatgtt gtgtgcatat ttttttttgt gcatgcacat gcatgtaaat
      721 agctaaaatt atgaacattt tattttttgt tcagaaaaaa aaaactttac acacataaaa
      781 tggctagtat gaatagccat attttatata aattaaatcc tatgaattta tgaccatatt
      841 aaaaatttag atatttatgg aacataatat gtttgaaaca ataagacaaa attattatta
      901 ttattattat ttttactgtt ataattatgt tgtctcttca atgattcata aatagttgga
      961 cttgattttt aaaatgttta taatatgatt agcatagtta aataaaaaaa gttgaaaaat
     1021 taaaaaaaaa catataaaca caaatgatgt tttttccttc aatttctcga gagcttaatt
     1081 cttttcgagc tctttatgct taagtttaca atttaatatt catactttaa gtattttttg
     1141 tagtatccta gatattgtgc tttaaatgct cacccctcaa agcaccagta atattttcat
     1201 ccactgaaat accattaaat tttcaaaaaa atactatgca tataatgtta tacatataaa
     1261 cataaaacgc catgtaaatc aaaaaatata taaaaatatg tataaaaata aatatgcact
     1321 aaatataagc taattatgca taaaaattaa agtgcccttt attaactagc tagtcgtaat
     1381 tatttatatt tctatgttat aaaaaaatcc tcatataata atataattaa tatatgtaat
     1441 gtttttttta ttttataatt ttaatataaa ataatatgta aattaattca aaaaataaat
     1501 ataattgttg tgaaacaaaa aacgtaattt tttcatttgc cttcaaaatt taaatttatt
     1561 ttaatatttc ctaaaatata tatactttgt gtataaatat ataaaaatat atatttgctt
     1621 ataaataaat aaaaatttta taaaaatgac tagtatgtac ccatacgatg ttccagatta
     1681 cgcttcgccg aagaaaaagc gcaaggtcga agcgtccgac aagaagtaca gcatcggcct
     1741 ggccatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag
     1801 caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg
     1861 agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag
     1921 aagaagatac accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga
     1981 gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga
     2041 ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca
     2101 cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc
     2161 cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct
     2221 gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt
     2281 gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa
     2341 ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct
     2401 gcccggcgag aagaagaatg gcctgttcgg caacctgatt gccctgagcc tgggcctgac
     2461 ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga
     2521 cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct
     2581 gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa
     2641 caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca
     2701 ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga
     2761 gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca
     2821 ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact
     2881 gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag
     2941 catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt
     3001 ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc
     3061 ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag
     3121 cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca
     3181 gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc
     3241 caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata
     3301 cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt
     3361 ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt
     3421 caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc
     3481 ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa
     3541 tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag
     3601 agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa
     3661 gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg
     3721 catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc
     3781 caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca
     3841 gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg
     3901 cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa
     3961 agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac
     4021 cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa
     4081 agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga
     4141 gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga
     4201 catcaaccgg ctgtccgact acgatgtgga cgccatcgtg cctcagagct ttctgaagga
     4261 cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa
     4321 cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc
     4381 caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag
     4441 cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa
     4501 gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct
     4561 gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga
     4621 tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct
     4681 gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt
     4741 gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat
     4801 cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga
     4861 gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac
     4921 cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat
     4981 gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc
     5041 tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa
     5101 gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt
     5161 ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat
     5221 ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga
     5281 agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg
     5341 ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc
     5401 ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc
     5461 cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat
     5521 catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa
     5581 agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat
     5641 catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac
     5701 caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca
     5761 ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacag
     5821 ccccaagaag aagagaaagg tggaggccag cgggtaggaa ttcgaaagat cccaacgaaa
     5881 agagagacca catggtcctt cttgagtttg taacagctgc tgggattaca catggcatgg
     5941 atgaactata caaataagga tccgtttttc ttacttatat atttatacca attgattgta
     6001 tttataactg taaaaatgtg tatgttgtgt gcatattttt ttttgtgcat gcacatgcat
     6061 gtaaatagct aaaattatga acattttatt ttttgttcag aaaaaaaaaa ctttacacac
     6121 ataaaatggc tagtatgaat agccatattt tatataaatt aaatcctatg aatttatgac
     6181 catattaaaa atttagatat ttatggaaca taatatgttt gaaacaataa gacaaaatta
     6241 ttattattat tattattttt actgttataa ttatgttgtc tcttcaatga ttcataaata
     6301 gttggacttg atttttaaaa tgtttataat atgattagca tagttaaata aaaaaagttg
     6361 aaaaattaaa aaaaaacata taaacacaaa tgatgttttt tccttcaatt tcggcgcctg
     6421 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     6481 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     6541 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     6601 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     6661 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     6721 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     6781 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     6841 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     6901 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     6961 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     7021 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     7081 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     7141 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     7201 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     7261 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     7321 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     7381 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     7441 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     7501 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     7561 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     7621 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     7681 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     7741 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     7801 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     7861 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     7921 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     7981 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     8041 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     8101 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     8161 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     8221 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     8281 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     8341 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     8401 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     8461 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     8521 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     8581 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     8641 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     8701 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     8761 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     8821 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     8881 tacgccaagc ttgataattc ctgcagccca gcttaattct tttcgagctc tttatgctta
     8941 agtttacaat ttaatattca tactttaagt attttttgta gtatcctaga tattgtgctt
     9001 taaatgctca cccctcaaag caccagtaat attttcatcc actgaaatac cattaaattt
     9061 tcaaaaaaat actatgcata taatgttata catataaaca taaaacgcca tgtaaatcaa
     9121 aaaatatata aaaatatgta taaaaataaa tatgcactaa atataagcta attatgcata
     9181 aaaattaaag tgccctttat taactagcta gtcgtaatta tttatatttc tatgttataa
     9241 aaaaatcctc atataataat ataattaata tatgtaatgt tttttttatt ttataatttt
     9301 aatataaaat aatatgtaaa ttaattcaaa aaataaatat aattgttgtg aaacaaaaaa
     9361 cgtaattttt tcatttgcct tcaaaattta aatttatttt aatatttcct aaaatatata
     9421 tactttgtgt ataaatatat aaaaatatat atttgcttat aaataaataa aaaattttat
     9481 aaaacatagg gggatccatg gttggttcgc taaactgcat cgtcgctgtg tcccagaaca
     9541 tgggcatcgg caagaacggg gacctgccct ggccaccgct caggaacgaa tttagatatt
     9601 tccagagaat gaccacaacc tcttcagtag aaggtaaaca gaatctggtg attatgggta
     9661 agaagacctg gttctccatt cctgagaaga atcgaccttt aaagggtaga attaatttag
     9721 ttctcagcag agaactcaag gaacctccac aaggagctca ttttctttcc agaagtctag
     9781 atgatgcctt aaaacttact gaacaaccag aattagcaaa taaagtagac atggtctgga
     9841 tagttggtgg cagttctgtt tataaggaag ccatgaatca cccaggccat cttaaactat
     9901 ttgtgacaag gatcatgcaa gactttgaaa gtgacacgtt ttttccagaa attgatttgg
     9961 agaaatataa acttctgcca gaatacccag gtgttctctc tgatgtccag gaggagaaag
    10021 gcattaagta caaatttgaa gtatatgaga agaatgatta aggatcccgt ttttcttact
    10081 tatatattta taccaattga ttgtatttat aactgtaaaa atgtgtatgt tgtgtgcata
    10141 tttttttttg tgcatgcaca tgcatgtaaa tagctaaaat tatgaacatt ttattttttg
    10201 ttcagaaaaa aaaactttac acacataaaa tggctagtat gaatagccat attttatata
    10261 aattaaatcc tatgaattta tgaccatatt aaaaatttag atatttatgg aacataatat
    10321 gtttgaaaca ataagacaaa attattatta ttattattat ttttactgtt ataattatgt
    10381 tgtctcttca atgattcata aatagttgga cttgattttt aaaatgttta taatatgatt
    10441 agcatagtta aataaaaaaa gttgaaaaat taaaaaaaaa catataaaca caaatgatgt
    10501 tttttccttc aatttcgatg
//
