LOCUS       sequence-349296-        9255 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SpCas9 and sgRNA. Can be integrated into the genome upon 
            co-delivery of Sleeping Beauty transposase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9255)
  AUTHORS   Kaltenbacher T, Loprich J, Maresch R, Weber J, Muller S, Oellinger 
            R, Gross N, Griger J, de Andrade Kratzig N, Avramopoulos P, 
            Ramanujam D, Brummer S, Widholz SA, Barthel S, Falcomata C, Pfaus A,
            Alnatsha A, Mayerle J, Schmidt-Supprian M, Reichert M, Schneider G, 
            Ehmer U, Braun CJ, Saur D, Engelhardt S, Rad R
  TITLE     CRISPR somatic genome engineering and cancer modeling in the mouse 
            pancreas and liver.
  JOURNAL   Nat Protoc. 2022 Apr;17(4):1142-1188. doi: 
            10.1038/s41596-021-00677-0. Epub 2022 Mar 14.
  PUBMED    35288718
REFERENCE   2  (bases 1 to 9255)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41596-021-00677-0"; journalName: "Nat Protoc"; date: 
            "2022-04"; volume: "17"; issue: "4"; pages: "1142-1188"
COMMENT     Sequence Label: CRISPR-SB
FEATURES             Location/Qualifiers
     source          1..9255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        100..348
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     100..120
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     271..290
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        367..442
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        539..824
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          1075..1302
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1314..1323
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1323..1388
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1395..1415
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1440..5540
                     /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             1440..5540
                     /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             5541..5588
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5616..5633)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5622..5829
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6240..6380
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6240..6369
                     /label=AAV2 ITR
     rep_origin      6455..6910
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6542..6561)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6752..6773
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6927..6946)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7046..7068
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7106..7124)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7192..7296
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7297..7365,7366..8157)
                     /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(7515..7534)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8328..8916
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8817..8836
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaaaatgatg tcatggcttt agaagcttct gatagactaa ttgacatcat ttgagtcaat
       61 tggaggtgta cctgtggatg tatttcaagg cctacatgtg agggcctatt tcccatgatt
      121 ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat taatttgact
      181 gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt tcttgggtag
      241 tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt aacttgaaag
      301 tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgg gtcttcgaga
      361 agacctgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
      421 aaaagtggca ccgagtcggt gcttttttgt tttagagcta gaaatagcaa gttaaaataa
      481 ggctagtccg tttttagcgc gtgcgccaat tctgcagaca aatggctcta gaggtacccg
      541 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      601 cgtcaatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      661 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      721 tcaatgacgg taaatggccc gcctggcatt gtgcccagta catgacctta tgggactttc
      781 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca
      841 cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg tatttattta
      901 ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggggg gcggggcggg
      961 gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt
     1021 atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc
     1081 gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg
     1141 ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc
     1201 tgtaattagc tgagcaagag gtaagggttt aagggatggt tggttggtgg ggtattaatg
     1261 tttaattacc tggagcacct gcctgaaatc actttttttc aggttggacc ggtgccacca
     1321 tggactataa ggaccacgac ggagactaca aggatcatga tattgattac aaagacgatg
     1381 acgataagat ggccccaaag aagaagcgga aggtcggtat ccacggagtc ccagcagccg
     1441 acaagaagta cagcatcggc ctggacatcg gcaccaactc tgtgggctgg gccgtgatca
     1501 ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca
     1561 gcatcaagaa gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca
     1621 cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc
     1681 tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg
     1741 aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca
     1801 tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac
     1861 tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga
     1921 tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg
     1981 acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca
     2041 acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc
     2101 tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggaaacctga
     2161 ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg
     2221 ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga
     2281 tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc
     2341 tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga
     2401 tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc
     2461 agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct
     2521 acattgacgg cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa
     2581 agatggacgg caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc
     2641 agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca
     2701 ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga
     2761 agatcctgac cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat
     2821 tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg
     2881 tggacaaggg cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc
     2941 tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata
     3001 acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg
     3061 gcgagcagaa aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga
     3121 agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg
     3181 gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca
     3241 aggacaagga cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga
     3301 ccctgacact gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc
     3361 tgttcgacga caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc
     3421 tgagccggaa gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt
     3481 tcctgaagtc cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc
     3541 tgacctttaa agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg
     3601 agcacattgc caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga
     3661 aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg
     3721 aaatggccag agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga
     3781 agcggatcga agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg
     3841 aaaacaccca gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata
     3901 tgtacgtgga ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gaccatatcg
     3961 tgcctcagag ctttctgaag gacgactcca tcgacaacaa ggtgctgacc agaagcgaca
     4021 agaaccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact
     4081 actggcggca gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca
     4141 aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg
     4201 tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta
     4261 agtacgacga gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc
     4321 tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc
     4381 accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc
     4441 ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga
     4501 tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca
     4561 tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc
     4621 tgatcgagac aaacggcgaa accggggaga tcgtgtggga taagggccgg gattttgcca
     4681 ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga
     4741 caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgataag ctgatcgcca
     4801 gaaagaagga ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt
     4861 ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag
     4921 agctgctggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc
     4981 tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact
     5041 ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccggc gaactgcaga
     5101 agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact
     5161 atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc
     5221 acaagcacta cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc
     5281 tggccgacgc taatctggac aaagtgctgt ccgcctacaa caagcaccgg gataagccca
     5341 tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg
     5401 ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc accaaagagg
     5461 tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc
     5521 tgtctcagct gggaggcgac aaaaggccgg cggccacgaa aaaggccggc caggcaaaaa
     5581 agaaaaagta agaattccta gagctcgctg atcagcctcg actgtgcctt ctagttgcca
     5641 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     5701 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     5761 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaga atagcaggca
     5821 tgctggggag cggccgcaca cattccacag aattccttga aatacatcca caggtacacc
     5881 tccaattgac tcaaatgatg tcaattagtc tatcagaagc ttctaaagcc atgacatcat
     5941 tttctggaat tttccaagct gtttaaaggc acagtcaact tagtgtatgt aaacttctga
     6001 cccactggaa ttgtgataca gtgaattata agtgaaataa tctgtctgta aacaattgtt
     6061 ggaaaaatga cttgtgtcat gcacaaagta gatgtcctaa ctgacttgcc aaaactattg
     6121 tttgttaaca agaaatttgt ggagtagttg aaaaacgagt tttaatgact ccaacttaag
     6181 tgtatgtaaa cttccgactt caactgtagg gtaccgagct cgaattcact ggcggccgca
     6241 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     6301 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     6361 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     6421 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     6481 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     6541 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     6601 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     6661 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     6721 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     6781 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     6841 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     6901 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     6961 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     7021 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     7081 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     7141 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     7201 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     7261 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     7321 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     7381 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     7441 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     7501 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     7561 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     7621 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     7681 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     7741 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     7801 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     7861 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     7921 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     7981 attgctgata aatctggagc cggtgagcgt ggaagccgcg gtatcattgc agcactgggg
     8041 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     8101 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     8161 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     8221 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     8281 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     8341 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     8401 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     8461 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     8521 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     8581 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     8641 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     8701 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     8761 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     8821 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     8881 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     8941 cggttcctgg ccttttgctg gccttttgct cacatgtcga gctcggtacc ctacagttga
     9001 agtcggaagt ttacatacac ttaagttgga gtcattaaaa ctcgtttttc aactactcca
     9061 caaatttctt gttaacaaac aatagttttg gcaagtcagt taggacatct actttgtgca
     9121 tgacacaagt catttttcca acaattgttt acagacagat tatttcactt ataattcact
     9181 gtatcacaat tccagtgggt cagaagttta catacactaa gttgactgtg cctttaaaca
     9241 gcttggaaaa ttcca
//
