LOCUS       sequence-417236-       12623 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR/Rad52-Cas9 expression vector for Animal gene editing, 
            harboring the ScRad52-SpCas9 and a VEGFA.gRNA expression cassette. 
            Key Construct: hU6-VEGFa.gRNA-CMV-ScRad52-SpCas9..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12623)
  AUTHORS   Shao S, Ren C, Liu Z, Bai Y, Chen Z, Wei Z, Wang X, Zhang Z, Xu K
  TITLE     Enhancing CRISPR/Cas9-mediated homology-directed repair in mammalian
            cells by expressing Saccharomyces cerevisiae Rad52.
  JOURNAL   Int J Biochem Cell Biol. 2017 Nov;92:43-52. doi: 
            10.1016/j.biocel.2017.09.012. Epub 2017 Sep 18.
  PUBMED    28928041
REFERENCE   2  (bases 1 to 12623)
  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: "Int J 
            Biochem Cell Biol."; date: "2017-11"; volume: "92:43-52. doi"; 
            pages: " 10.1016/j.biocel.2017.09.012. Epub 2017 Sep 18"
COMMENT     Sequence Label: pScRad52-SpCas9 (pXK-1646)
FEATURES             Location/Qualifiers
     source          1..12623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     214..233
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        312..387
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        401..686
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        688..962
                     /label=chicken beta-actin promoter
     intron          963..1190
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1202..1211
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1211..1276
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1283..1303
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2729..6829
                     /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             2729..6829
                     /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"
     protein_bind    complement(6855..6888)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    6944..7532
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6997..7017)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7415..7426)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(7535..7551)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7536..7552)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8061..8241
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(8303..8891)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8383..8402)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9062..9853,9854..9922))
                     /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     9685..9704
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9923..10027)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10102..10121)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        10293..10672
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        10674..10872
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     10827..10847
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             10890..11070
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11117..11242
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    11739..11972
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             12157..12201
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    12499..12616
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 attacaggga cagcagagat ccagtttggt tagtaccggg cccgctctag atttcccatg
       61 attccttcat atttgcatat acgatacaag gctgttagag agataattgg aattaatttg
      121 actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg
      181 tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga
      241 aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac cctcggccac
      301 cacagggaag cgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa
      361 cttgaaaaag tggcaccgag tcggtgcttt tttgcggccg cgttacataa cttacggtaa
      421 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata gtaacgccaa
      481 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      541 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      601 ccgcctggca ttgtgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      661 acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc
      721 ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg
      781 cagcgatggg ggcggggggg ggggggggcg cgccaggcgg ggcggggcgg ggcgaggggc
      841 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      901 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      961 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
     1021 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
     1081 ctccgggctg taattagctg agcaagaggt aagggtttaa gggatggttg gttggtgggg
     1141 tattaatgtt taattacctg gagcacctgc ctgaaatcac tttttttcag gttggaccgg
     1201 tgccaccatg gactataagg accacgacgg agactacaag gatcatgata ttgattacaa
     1261 agacgatgac gataagatgg ccccaaagaa gaagcggaag gttgctagaa tgaatgaaat
     1321 tatggatatg gatgagaaga agcccgtttt cggtaaccat tccgaggaca tacagaccaa
     1381 attggacaag aaattaggac ctgagtatat ctccaagaga gttgggtttg gaacaagcag
     1441 gattgcatac atcgaaggtt ggagagtaat taatcttgct aatcaaatct ttggatataa
     1501 tggctggtct acggaggtaa aaagtgtagc tatcgatttt ttggatgagc gacagggaaa
     1561 gtttagcata gggtgtactg caattgttcg tgttacgttg actagcggga cttataggga
     1621 agatattggg tatggtaccg tggagaacga aagacggaga cctgccgctt tcgaaagggc
     1681 caagaaatct gccgttacgg atgccttgaa aagatctttg agagggtttg gtaatgctct
     1741 aggaaattgt ctttacgata aagattttct ggcaaagatc gataaggtga agttcgatcc
     1801 gccagatttt gacgaaaaca atttgtttag gccaacggat gaaatcagcg aaagctcaag
     1861 aacgaatact ttgcatgaaa atcaagaaca acaacaatac ccgaataaaa gaaggcaatt
     1921 gactaaagtt acaaatacca atcccgactc gacgaagaac ctggtgaaaa tagaaaatac
     1981 agtaagtcga ggtaccccaa tgatggccgc acctgctgag gctaacagta aaaacagcag
     2041 taataaagat actgatctca aatctctcga tgcttccaaa caggatcaag atgaccttct
     2101 cgatgattct cttatgttta gcgatgattt tcaagacgac gacttgataa atatgggcaa
     2161 cacaaacagt aatgttttaa cgacagagaa ggatcccgtt gtagctaagc aaagcccaac
     2221 tgctagctca aacccagaga cagaacaaat cacctttgtt acagctaagg ttgcaacttc
     2281 cgtacaaaat gaacgttata ttggtgaaga gagcattttt gatcccaaat accaggcaca
     2341 atccattagg cacaccgttg atcagactac gtccaagcat gttccagcga gtgtgctaaa
     2401 agataaaacg atgaccaccg cgagggattc tgtctatgaa aaatttgcac caaagggaaa
     2461 acaattgagc atgaagaaca atgataaaga actggggcct catatgccag aaggggctgg
     2521 gaatcaagta ccgcgtgaaa ccacaccaat taaaacgaac gctacagctt ttccgcctgc
     2581 agctgctcca cggtttgcgc cgccttctaa ggtagtacat cctaatggaa atggtgcagt
     2641 gccagccctt ccgcaacaaa gatcgacacg aagagaagtt ggaagaccaa agatcaatcc
     2701 cctgcatgca cgcaagccta ctcgatccga caagaagtac agcatcggcc tggacatcgg
     2761 caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt
     2821 caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg gagccctgct
     2881 gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata
     2941 caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa
     3001 ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa
     3061 gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta
     3121 ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg
     3181 gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg
     3241 cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta
     3301 caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca aggccatcct
     3361 gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga
     3421 gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga cccccaactt
     3481 caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga
     3541 cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc
     3601 cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat
     3661 caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct
     3721 gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt
     3781 cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc aggaagagtt
     3841 ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa
     3901 gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca
     3961 ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt
     4021 cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt
     4081 gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac
     4141 catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat
     4201 cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag
     4261 cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga
     4321 gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct
     4381 gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat
     4441 cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg
     4501 cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa
     4561 cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat
     4621 cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa
     4681 gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga
     4741 caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa
     4801 cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca
     4861 ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc
     4921 cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg
     4981 ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa
     5041 gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg
     5101 cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta
     5161 cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg
     5221 gctgtccgac tacgatgtgg accatatcgt gcctcagagc tttctgaagg acgactccat
     5281 cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca acgtgccctc
     5341 cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat
     5401 tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga
     5461 taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc
     5521 acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga
     5581 agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt
     5641 ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt
     5701 cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga
     5761 ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc
     5821 taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct
     5881 ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat
     5941 cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt
     6001 gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc
     6061 caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg
     6121 cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg
     6181 caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag
     6241 cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa
     6301 ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag
     6361 aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata
     6421 tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa
     6481 tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca
     6541 gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc
     6601 cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct
     6661 gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga
     6721 ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat
     6781 caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgacc tcgagcaatt
     6841 cgtcgaggga cctaataact tcgtatagca tacattatac gaagttatac atgtttaagg
     6901 gttccggttc cactaggtac aattcgatat caagcttatc gataatcaac ctctggatta
     6961 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     7021 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     7081 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     7141 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     7201 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     7261 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     7321 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     7381 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     7441 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     7501 gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gatcgagacc
     7561 tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct gattgtgcct
     7621 ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag gtacctttaa
     7681 gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac
     7741 tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg atctaccaca
     7801 cacaaggcta cttccctgat tggcagaact acacaccagg gccagggatc agatatccac
     7861 tgacctttgg atggtgctac aagctagtac cagttgagca agagaaggta gaagaagcca
     7921 atgaaggaga gaacacccgc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
     7981 agagagaagt attagagtgg aggtttgaca gccgcctagc atttcatcac atggcccgag
     8041 agctgcatcc ggactgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     8101 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     8161 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     8221 cagtgtggaa aatctctagc agcatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     8281 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     8341 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     8401 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     8461 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     8521 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     8581 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     8641 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     8701 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     8761 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     8821 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     8881 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     8941 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     9001 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     9061 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     9121 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     9181 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     9241 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     9301 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     9361 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     9421 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     9481 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     9541 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     9601 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     9661 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     9721 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     9781 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     9841 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     9901 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     9961 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
    10021 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
    10081 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
    10141 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
    10201 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
    10261 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
    10321 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
    10381 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
    10441 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
    10501 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
    10561 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
    10621 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
    10681 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
    10741 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
    10801 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
    10861 ctatataagc agcgcgtttt gcctgtactg ggtctctctg gttagaccag atctgagcct
    10921 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
    10981 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
    11041 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
    11101 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
    11161 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
    11221 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
    11281 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
    11341 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
    11401 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
    11461 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
    11521 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
    11581 ggccggccgc tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa
    11641 ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag
    11701 agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc
    11761 ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga
    11821 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
    11881 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
    11941 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
    12001 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
    12061 tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata
    12121 cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa
    12181 ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata
    12241 aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt
    12301 tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca
    12361 accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga
    12421 gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat
    12481 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
    12541 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
    12601 aaaaattcaa aattttcggg ttt
//
