LOCUS       sequence-267599-        9656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for sgRNAs cloned into the BsaI sites and for 
            expression of Cas9 with 126aa MRN-recruiting domain from HSV-1 UL12 
            fused to C-terminus of Cas9, linked to mCherry via a T2A peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9656)
  AUTHORS   Reuven N, Adler J, Broennimann K, Myers N, Shaul Y
  TITLE     Recruitment of DNA Repair MRN Complex by Intrinsically Disordered 
            Protein Domain Fused to Cas9 Improves Efficiency of CRISPR-Mediated 
            Genome Editing.
  JOURNAL   Biomolecules. 2019 Oct 8;9(10). pii: biom9100584. doi: 
            10.3390/biom9100584.
  PUBMED    31597252
REFERENCE   2  (bases 1 to 9656)
  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: 
            "Biomolecules."; date: "2019-10-8"; volume: "9(10). pii: 
            biom9100584. doi"; pages: " 10.3390/biom9100584"
COMMENT     Sequence Label: pU6-(BsaI)_CBh-UC-Cas9-T2A-mCherry
FEATURES             Location/Qualifiers
     source          1..9656
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          275..502
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      514..523
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             523..588
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             595..615
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             640..4740
                     /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             640..4740
                     /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             5170..5223
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             5230..5940
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5375..5393)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(5965..5982)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5971..6178
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6187..6327
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6187..6316
                     /label=AAV2 ITR
     rep_origin      6402..6857
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6489..6508)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6699..6720
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6874..6893)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6993..7015
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7053..7071)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7139..7243
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7244..7312,7313..8104)
                     /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(7462..7481)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8275..8863
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8764..8783
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8925..9173
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8925..8945
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9096..9115
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        9194..9269
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        9366..9651
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        join(9653..9656,1..274)
                     /label=chicken beta-actin promoter
ORIGIN
        1 ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt
       61 gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg ggggggggcg
      121 cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc
      181 ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg
      241 gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc
      301 cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact
      361 cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc tgagcaagag
      421 gtaagggttt aagggatggt tggttggtgg ggtattaatg tttaattacc tggagcacct
      481 gcctgaaatc actttttttc aggttggacc ggtgccacca tggactataa ggaccacgac
      541 ggagactaca aggatcatga tattgattac aaagacgatg acgataagat ggccccaaag
      601 aagaagcgga aggtcggtat ccacggagtc ccagcagccg acaagaagta cagcatcggc
      661 ctggacatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc
      721 agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc
      781 ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc
      841 agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac
      901 gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa
      961 gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac
     1021 cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag
     1081 gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc
     1141 ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg
     1201 gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc
     1261 aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag
     1321 ctgcccggcg agaagaagaa tggcctgttc ggaaacctga ttgccctgag cctgggcctg
     1381 acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag
     1441 gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac
     1501 ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg
     1561 aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac
     1621 caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa
     1681 gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc
     1741 caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa
     1801 ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc
     1861 agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat
     1921 ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc
     1981 ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag
     2041 agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc
     2101 cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg
     2161 cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa
     2221 tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc
     2281 gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac
     2341 ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac
     2401 gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac
     2461 aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac
     2521 agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg
     2581 aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac
     2641 ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc
     2701 gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc
     2761 cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc
     2821 ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg
     2881 aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag
     2941 accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc
     3001 aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac
     3061 gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg
     3121 gacatcaacc ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag
     3181 gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac
     3241 aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac
     3301 gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg
     3361 agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca
     3421 aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag
     3481 ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag
     3541 gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac
     3601 ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc
     3661 gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa
     3721 atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc
     3781 gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa
     3841 accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc
     3901 atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag
     3961 tctatcctgc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct
     4021 aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa
     4081 gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc
     4141 atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa
     4201 gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac
     4261 ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg
     4321 ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc
     4381 cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag
     4441 atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac
     4501 aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat
     4561 atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac
     4621 accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc
     4681 caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac
     4741 aaaaggccgg cggccacgaa aaaggccggc caggcaaaaa agaaaatgga gtccacggga
     4801 ggcccagcat gtccgccggg acgcaccgtg actaagcgtt cctgggccct ggccgaggac
     4861 acccctcgtg gccccgacag cccccccaag cgcccccgcc ctaacagtct tccgctaaca
     4921 accaccttcc gtcccctgcc ccccccaccc cagacgacgt cagctgtgga cccgagctcc
     4981 cattcgcccg ttaacccccc acgtgatcag cacgccaccg acaccgcaga cgaaaagccc
     5041 cgggccgcgt cgccggcact ttctgacgcc tcagggcctc cgaccccaga cattccgcta
     5101 tctcctgggg gcacccacgc ccgcgacccg gacgccgatc ccgactcccc ggaccttgac
     5161 tctaagcttg agggcagagg aagtctgcta acatgcggtg acgtggagga gaatcccggc
     5221 cctgctagca tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg
     5281 cgcttcaagg tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag
     5341 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc
     5401 cccctgccct tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac
     5461 gtgaagcacc ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag
     5521 tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc
     5581 ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctcagac
     5641 ggccccgtaa tgcagaagaa aaccatgggc tgggaggcct cctccgagcg gatgtacccc
     5701 gaggacggcg ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac
     5761 tacgacgctg aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc
     5821 tacaacgtca acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa
     5881 cagtacgaac gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa
     5941 gaattcctag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     6001 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     6061 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     6121 gggtggggca ggacagcaag ggggaggatt gggaagagaa tagcaggcat gctggggagc
     6181 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     6241 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     6301 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     6361 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     6421 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     6481 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     6541 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     6601 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     6661 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     6721 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     6781 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     6841 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     6901 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     6961 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     7021 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     7081 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     7141 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     7201 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     7261 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     7321 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     7381 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     7441 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     7501 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     7561 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     7621 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     7681 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     7741 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     7801 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     7861 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     7921 ctggtttatt gctgataaat ctggagccgg tgagcgtgga agccgcggta tcattgcagc
     7981 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     8041 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     8101 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     8161 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     8221 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     8281 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     8341 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     8401 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     8461 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     8521 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     8581 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     8641 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     8701 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     8761 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     8821 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     8881 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtgagggc ctatttccca
     8941 tgattccttc atatttgcat atacgataca aggctgttag agagataatt ggaattaatt
     9001 tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg
     9061 ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt
     9121 gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac accggagacc
     9181 tgtacaggtc tctgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
     9241 aacttgaaaa agtggcaccg agtcggtgct tttttgtttt agagctagaa atagcaagtt
     9301 aaaataaggc tagtccgttt ttagcgcgtg cgccaattct gcagacaaat ggctctagag
     9361 gtacccgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     9421 ccattgacgt caatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     9481 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     9541 attgacgtca atgacggtaa atggcccgcc tggcattgtg cccagtacat gaccttatgg
     9601 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtcga
//
