LOCUS       sequence-273439-        8734 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SpCas9 and sgRNA Scaffold.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8734)
  AUTHORS   Lu Z, Yao X, Lyu P, Yadav M, Yoo K, Atala A, Lu B
  TITLE     Lentiviral Capsid-Mediated Streptococcus pyogenes Cas9 
            Ribonucleoprotein Delivery for Efficient and Safe Multiplex Genome 
            Editing.
  JOURNAL   CRISPR J. 2021 Mar 16. doi: 10.1089/crispr.2020.0106.
  PUBMED    33733873
REFERENCE   2  (bases 1 to 8734)
  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: "CRISPR J. 
            2021 Mar 16. doi: 10.1089/crispr.2020.0106."
COMMENT     Sequence Label: pspCas9-3'UTR-ST2-com-vector
FEATURES             Location/Qualifiers
     source          1..8734
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        77..353
                     /label=chicken beta-actin promoter
     intron          354..581
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      593..602
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             602..667
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             674..694
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             719..4819
                     /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             719..4819
                     /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             4820..4867
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5177..5194)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5183..5390
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5402..5542
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5402..5531
                     /label=AAV2 ITR
     rep_origin      5617..6072
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5704..5723)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5914..5935
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6089..6108)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6208..6230
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6268..6286)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6354..6458
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6459..6527,6528..7319)
                     /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(6677..6696)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7490..8078
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7979..7998
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8140..8388
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8140..8160
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8311..8330
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        join(8524..8734,1..75)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 tgtgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
       61 atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc
      121 ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg
      181 gcgggggggg gggggggsgc gcgccaggcg gggcggggcg gggcgagggg cggggcgggg
      241 cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta
      301 tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg
      361 ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc
      421 tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct
      481 gtaattagct gagcaagagg taagggttta agggatggtt ggttggtggg gtattaatgt
      541 ttaattacct ggagcacctg cctgaaatca ctttttttca ggttggaccg gtgccaccat
      601 ggactataag gaccacgacg gagactacaa ggatcatgat attgattaca aagacgatga
      661 cgataagatg gccccaaaga agaagcggaa ggtcggtatc cacggagtcc cagcagccga
      721 caagaagtac agcatcggcc tggacatcgg caccaactct gtgggctggg ccgtgatcac
      781 cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag
      841 catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac
      901 ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct
      961 gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga
     1021 agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat
     1081 cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact
     1141 ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat
     1201 caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga
     1261 caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa
     1321 cgccagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct
     1381 ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat
     1441 tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc
     1501 caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat
     1561 cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct
     1621 gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat
     1681 caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca
     1741 gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta
     1801 cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa
     1861 gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca
     1921 gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat
     1981 tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa
     2041 gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt
     2101 cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt
     2161 ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct
     2221 gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa
     2281 cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg
     2341 cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa
     2401 gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg
     2461 cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa
     2521 ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac
     2581 cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct
     2641 gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct
     2701 gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt
     2761 cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct
     2821 gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga
     2881 gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa
     2941 ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga
     3001 aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa
     3061 gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga
     3121 aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat
     3181 gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt
     3241 gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa
     3301 gaaccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta
     3361 ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa
     3421 ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt
     3481 ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa
     3541 gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct
     3601 ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca
     3661 ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc
     3721 taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat
     3781 cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat
     3841 catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct
     3901 gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac
     3961 cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac
     4021 aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag
     4081 aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc
     4141 tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga
     4201 gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct
     4261 ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc
     4321 cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa
     4381 gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta
     4441 tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca
     4501 caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct
     4561 ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat
     4621 cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc
     4681 cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt
     4741 gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct
     4801 gtctcagctg ggaggcgaca aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa
     4861 gaaaaaggga tcctaagctc gctttcttgc tgtccaattt ctattaaagg ttcctttgtt
     4921 ccctaagtcc aactactaaa ctgggggata ttatgaaggg ccttgagcat ctggattctg
     4981 cctaataaaa aacatttatt ttcattgcta gctcgctttc ttgctgtcca atttctatta
     5041 aaggttcctt tgttccctaa gtccaactac taaactgggg gatattatga agggccttga
     5101 gcatctggat tctgcctaat aaaaaacatt tattttcatt gcgaattcta gaaaattcct
     5161 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     5221 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     5281 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     5341 caggacagca agggggagga ttgggaagag aatagcaggc atgctgggga ggtacggccg
     5401 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     5461 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     5521 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     5581 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     5641 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     5701 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     5761 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     5821 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     5881 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     5941 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     6001 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     6061 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     6121 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     6181 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     6241 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     6301 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     6361 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     6421 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     6481 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     6541 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     6601 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     6661 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     6721 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     6781 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     6841 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     6901 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     6961 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     7021 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     7081 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     7141 ttattgctga taaatctgga gccggtgagc gtggaagccg cggtatcatt gcagcactgg
     7201 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     7261 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     7321 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     7381 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     7441 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     7501 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     7561 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     7621 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     7681 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     7741 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     7801 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     7861 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     7921 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     7981 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     8041 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     8101 tacggttcct ggccttttgc tggccttttg ctcacatgtg agggcctatt tcccatgatt
     8161 ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat taatttgact
     8221 gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag
     8281 tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag
     8341 tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgg tgtcttcctc
     8401 gaggaagacc cgtttgagag ctagaaatag caagttcaaa taaggctagt ccgttatcaa
     8461 cttggctgaa tgcctgcgag catcccaccc aagtggcacc gagtcggtgc tttttttggt
     8521 acccgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     8581 attgacgtca atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     8641 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     8701 tgacgtcaat gacggtaaat ggcccgcctg gcat
//
