LOCUS       sequence-267596-        9272 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression vector for sgRNAs cloned into the BsaI sites and for 
            expression 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 9272)
  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 9272)
  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-Cas9-T2A-mCherry
FEATURES             Location/Qualifiers
     source          1..9272
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          156..383
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      395..404
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             404..469
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             476..496
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             521..4621
                     /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
                     ATLXHQSITGLYETRIDLSQLGGD"
     CDS             4622..4669
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4673..4726
                     /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             4733..5443
                     /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(4878..4896)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(5468..5485)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5474..5681
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5690..5830
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5690..5819
                     /label=AAV2 ITR
     rep_origin      5905..6360
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5992..6011)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6202..6223
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6377..6396)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6496..6518
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6556..6574)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6642..6746
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6747..6815,6816..7607)
                     /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(6965..6984)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7778..8366
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8267..8286
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8428..8676
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8428..8448
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8599..8618
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8697..8772
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8869..9154
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 smgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg
       61 cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc
      121 ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc
      181 ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac
      241 tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag ctgagcaaga
      301 ggtaagggtt taagggatgg ttggttggtg gggtattaat gtttaattac ctggagcacc
      361 tgcctgaaat cacttttttt caggttggac cggtgccacc atggactata aggaccacga
      421 cggagactac aaggatcatg atattgatta caaagacgat gacgataaga tggccccaaa
      481 gaagaagcgg aaggtcggta tccacggagt cccagcagcc gacaagaagt acagcatcgg
      541 cctggacatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc
      601 cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat
      661 cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga agagaaccgc
      721 cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa
      781 cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga
      841 agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta
      901 ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa
      961 ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt
     1021 cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct
     1081 ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc
     1141 caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca
     1201 gctgcccggc gagaagaaga atggcctgtt cggaaacctg attgccctga gcctgggcct
     1261 gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa
     1321 ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga
     1381 cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt
     1441 gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca
     1501 ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa
     1561 agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag
     1621 ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga
     1681 actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg
     1741 cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga
     1801 tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat
     1861 cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa
     1921 gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc
     1981 ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct
     2041 gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa
     2101 atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat
     2161 cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta
     2221 cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa
     2281 cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga
     2341 caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga
     2401 cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat
     2461 gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa
     2521 cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt
     2581 cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat
     2641 ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc
     2701 cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt
     2761 gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca
     2821 gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat
     2881 caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa
     2941 cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact
     3001 ggacatcaac cggctgtccg actacgatgt ggaccatatc gtgcctcaga gctttctgaa
     3061 ggacgactcc atcgacaaca aggtgctgac cagaagcgac aagaaccggg gcaagagcga
     3121 caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcggc agctgctgaa
     3181 cgccaagctg attacccaga gaaagttcga caatctgacc aaggccgaga gaggcggcct
     3241 gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac
     3301 aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg agaatgacaa
     3361 gctgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa
     3421 ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta
     3481 cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg aaagcgagtt
     3541 cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga
     3601 aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac
     3661 cgagattacc ctggccaacg gcgagatccg gaagcggcct ctgatcgaga caaacggcga
     3721 aaccggggag atcgtgtggg ataagggccg ggattttgcc accgtgcgga aagtgctgag
     3781 catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga
     3841 gtctatcctg cccaagagga acagcgataa gctgatcgcc agaaagaagg actgggaccc
     3901 taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg tggtggccaa
     3961 agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat
     4021 catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa
     4081 agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa
     4141 cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg aactggccct
     4201 gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc
     4261 ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaagcact acctggacga
     4321 gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga
     4381 caaagtgctg tccgcctaca acaagcaccg ggataagccc atcagagagc aggccgagaa
     4441 tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca agtactttga
     4501 caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg ccaccctgay
     4561 ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggcga
     4621 caaaaggccg gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagc ttgagggcag
     4681 aggaagtctg ctaacatgcg gtgacgtgga ggagaatccc ggccctgcta gcatggtgag
     4741 caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat
     4801 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     4861 cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
     4921 ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga
     4981 catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     5041 cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
     5101 catctacaag gtgaagctgc gcggcaccaa cttcccctca gacggccccg taatgcagaa
     5161 gaaaaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa
     5221 gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa
     5281 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa
     5341 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
     5401 gggccgccac tccaccggcg gcatggacga gctgtacaag taagaattcc tagagctcgc
     5461 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     5521 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     5581 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     5641 aagggggagg attgggaaga gaatagcagg catgctgggg agcggccgca ggaaccccta
     5701 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     5761 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     5821 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     5881 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     5941 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg
     6001 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     6061 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     6121 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     6181 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta
     6241 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa
     6301 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     6361 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     6421 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     6481 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     6541 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     6601 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     6661 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     6721 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     6781 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     6841 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     6901 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     6961 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     7021 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     7081 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     7141 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     7201 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     7261 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     7321 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     7381 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     7441 aatctggagc cggtgagcgt ggaagccgcg gtatcattgc agcactgggg ccagatggta
     7501 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     7561 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     7621 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     7681 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     7741 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     7801 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     7861 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     7921 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     7981 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     8041 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     8101 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     8161 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     8221 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     8281 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     8341 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     8401 ccttttgctg gccttttgct cacatgtgag ggcctatttc ccatgattcc ttcatatttg
     8461 catatacgat acaaggctgt tagagagata attggaatta atttgactgt aaacacaaag
     8521 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
     8581 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
     8641 ttggctttat atatcttgtg gaaaggacga aacaccggag acctgtacag gtctctgttt
     8701 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
     8761 ccgagtcggt gcttttttgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg
     8821 tttttagcgc gtgcgccaat tctgcagaca aatggctcta gaggtacccg ttacataact
     8881 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaatagt
     8941 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     9001 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     9061 taaatggccc gcctggcatt gtgcccagta catgacctta tgggactttc ctacttggca
     9121 gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt
     9181 cactctcccc atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt
     9241 attttgtgca gcgatggggg cggggggggg gg
//
