LOCUS       sequence-177239-        8488 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  For transient expression of spCas9-nuclease and a sgRNA targeting 
            the mouse Rosa26 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8488)
  AUTHORS   Nora EP, Goloborodko A, Valton AL, Gibcus JH, Uebersohn A, Abdennur 
            N, Dekker J, Mirny LA, Bruneau BG
  TITLE     Targeted Degradation of CTCF Decouples Local Insulation of 
            Chromosome Domains from Genomic Compartmentalization.
  JOURNAL   Cell. 2017 May 18;169(5):930-944.e22. doi: 
            10.1016/j.cell.2017.05.004.
  PUBMED    28525758
REFERENCE   2  (bases 1 to 8488)
  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: 
            "10.1016/j.cell.2017.05"; journalName: "Cell"; date: "2017-05-18- 
            18"; volume: "169"; issue: "5"; pages: "930-944.e22"
COMMENT     Sequence Label: pX330-EN479
FEATURES             Location/Qualifiers
     source          1..8488
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          141..368
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      380..389
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             389..454
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             461..481
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             506..4606
                     /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             506..4606
                     /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             4607..4654
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4682..4699)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4688..4895
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4904..5044
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4904..5033
                     /label=AAV2 ITR
     rep_origin      5119..5574
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5206..5225)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5416..5437
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5591..5610)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5710..5732
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5770..5788)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5856..5960
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5961..6029,6030..6821)
                     /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(6179..6198)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6992..7580
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7481..7500
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7642..7890
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7642..7662
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7813..7832
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7912..7987
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8084..8369
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 cggggcgggg cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga
       61 gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa
      121 agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg
      181 ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg
      241 ggcgggacgg cccttctcct ccgggctgta attagctgag caagaggtaa gggtttaagg
      301 gatggttggt tggtggggta ttaatgttta attacctgga gcacctgcct gaaatcactt
      361 tttttcaggt tggaccggtg ccaccatgga ctataaggac cacgacggag actacaagga
      421 tcatgatatt gattacaaag acgatgacga taagatggcc ccaaagaaga agcggaaggt
      481 cggtatccac ggagtcccag cagccgacaa gaagtacagc atcggcctgg acatcggcac
      541 caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa
      601 ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt
      661 cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac
      721 cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt
      781 ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca
      841 cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc
      901 caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct
      961 gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga
     1021 cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa
     1081 ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc
     1141 tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa
     1201 gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa
     1261 gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga
     1321 cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc
     1381 caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac
     1441 caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac
     1501 cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga
     1561 ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta
     1621 caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct
     1681 gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca
     1741 gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct
     1801 gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg
     1861 ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat
     1921 caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga
     1981 gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct
     2041 gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg
     2101 aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt
     2161 caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga
     2221 gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac
     2281 ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga
     2341 ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga
     2401 ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg
     2461 gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa
     2521 gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt
     2581 catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt
     2641 gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat
     2701 taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg
     2761 gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg
     2821 acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag
     2881 ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct
     2941 gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct
     3001 gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg actccatcga
     3061 caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga
     3121 agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac
     3181 ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa
     3241 ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca
     3301 gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt
     3361 gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta
     3421 caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt
     3481 gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta
     3541 caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac
     3601 cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc
     3661 caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt
     3721 gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa
     3781 tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa
     3841 gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg
     3901 cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa
     3961 gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag
     4021 cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga
     4081 cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat
     4141 gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt
     4201 gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga
     4261 gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat
     4321 cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc
     4381 ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt
     4441 taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg
     4501 gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac
     4561 cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacaaaa ggccggcggc
     4621 cacgaaaaag gccggccagg caaaaaagaa aaagtaagaa ttcctagagc tcgctgatca
     4681 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     4741 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     4801 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     4861 gaggattggg aagagaatag caggcatgct ggggagcggc cgcaggaacc cctagtgatg
     4921 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     4981 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     5041 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     5101 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     5161 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     5221 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     5281 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     5341 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     5401 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     5461 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     5521 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     5581 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     5641 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     5701 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     5761 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     5821 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     5881 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     5941 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     6001 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     6061 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     6121 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     6181 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     6241 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     6301 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6361 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6421 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6481 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     6541 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     6601 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6661 gagccggtga gcgtggaagc cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6721 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6781 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     6841 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     6901 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     6961 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     7021 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     7081 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     7141 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     7201 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     7261 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7321 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7381 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7441 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     7501 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     7561 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     7621 gctggccttt tgctcacatg tgagggccta tttcccatga ttccttcata tttgcatata
     7681 cgatacaagg ctgttagaga gataattgga attaatttga ctgtaaacac aaagatatta
     7741 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta
     7801 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct
     7861 ttatatatct tgtggaaagg acgaaacacc gtgggcggga gtcttctggg cgttttagag
     7921 ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
     7981 tcggtgcttt tttgttttag agctagaaat agcaagttaa aataaggcta gtccgttttt
     8041 agcgcgtgcg ccaattctgc agacaaatgg ctctagaggt acccgttaca taacttacgg
     8101 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca atagtaacgc
     8161 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     8221 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     8281 ggcccgcctg gcattgtgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     8341 tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc
     8401 tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt
     8461 gtgcagcgat gggggcgggg gggggggg
//
