LOCUS       sequence-175360-        8319 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  To express sgRNA target HUWE1 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8319)
  AUTHORS   Xu Y, Anderson DE, Ye Y
  TITLE     The HECT domain ubiquitin ligase HUWE1 targets unassembled soluble 
            proteins for degradation.
  JOURNAL   Cell Discov. 2016 Nov 8;2:16040. eCollection 2016.
  PUBMED    27867533
REFERENCE   2  (bases 1 to 8319)
  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: "Cell 
            Discov."; date: "2016-11-8"; volume: "2"; pages: "16040. eCollection
            2016"
COMMENT     Sequence Label: HUWE1-sgRNA-1
FEATURES             Location/Qualifiers
     source          1..8319
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      224..233
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             233..298
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             305..325
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(350..373,374..376,377..4450)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             350..4450
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4451..4498
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4526..4543)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4532..4739
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4748..4888
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4748..4877
                     /label=AAV2 ITR
     rep_origin      4963..5418
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5050..5069)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5260..5281
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5435..5454)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5554..5576
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5614..5632)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5700..5804
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5805..5873,5874..6665)
                     /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(6023..6042)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6836..7424
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7325..7344
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7486..7734
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7486..7506
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7657..7676
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7756..7831
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        7928..8213
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga
       61 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
      121 tgagcaagag gtaagggttt aagggatggt tggttggtgg ggtattaatg tttaattacc
      181 tggagcacct gcctgaaatc actttttttc aggttggacc ggtgccacca tggactataa
      241 ggaccacgac ggagactaca aggatcatga tattgattac aaagacgatg acgataagat
      301 ggccccaaag aagaagcgga aggtcggtat ccacggagtc ccagcagccg acaagaagta
      361 cagcatcggc ctggccatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta
      421 caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa
      481 gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa
      541 gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat
      601 cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt
      661 cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga
      721 ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag
      781 caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg
      841 gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt
      901 catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg
      961 cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct
     1021 gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggaaacctga ttgccctgag
     1081 cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca
     1141 gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca
     1201 gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat
     1261 cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata
     1321 cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga
     1381 gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg
     1441 cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg
     1501 caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt
     1561 cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg
     1621 gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac
     1681 cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat
     1741 gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg
     1801 cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga
     1861 gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac
     1921 caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa
     1981 aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa
     2041 agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga
     2101 tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga
     2161 cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact
     2221 gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga
     2281 caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa
     2341 gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc
     2401 cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa
     2461 agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc
     2521 caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga
     2581 cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag
     2641 agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga
     2701 agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca
     2761 gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga
     2821 ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gaccatatcg tgcctcagag
     2881 ctttctgaag gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg
     2941 caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca
     3001 gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag
     3061 aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg
     3121 gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga
     3181 gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga
     3241 tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca
     3301 cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga
     3361 aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag
     3421 cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt
     3481 tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac
     3541 aaacggcgaa accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa
     3601 agtgctgagc atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt
     3661 cagcaaagag tctatcctgc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga
     3721 ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt
     3781 ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg
     3841 gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa
     3901 gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga
     3961 gctggaaaac ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga
     4021 actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct
     4081 gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta
     4141 cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc
     4201 taatctggac aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca
     4261 ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa
     4321 gtactttgac accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc
     4381 caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct
     4441 gggaggcgac aaaaggccgg cggccacgaa aaaggccggc caggcaaaaa agaaaaagta
     4501 agaattccta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     4561 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     4621 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     4681 ggggtggggc aggacagcaa gggggaggat tgggaagaga atagcaggca tgctggggag
     4741 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     4801 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     4861 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
     4921 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     4981 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct
     5041 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     5101 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     5161 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     5221 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     5281 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     5341 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     5401 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     5461 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     5521 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     5581 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     5641 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     5701 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     5761 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     5821 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5881 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     5941 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     6001 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     6061 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     6121 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     6181 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     6241 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     6301 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     6361 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     6421 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     6481 gctggtttat tgctgataaa tctggagccg gtgagcgtgg aagccgcggt atcattgcag
     6541 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6601 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6661 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     6721 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6781 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6841 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6901 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6961 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     7021 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     7081 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     7141 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     7201 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     7261 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     7321 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     7381 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     7441 cctttttacg gttcctggcc ttttgctggc cttttgctca catgtgaggg cctatttccc
     7501 atgattcctt catatttgca tatacgatac aaggctgtta gagagataat tggaattaat
     7561 ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt
     7621 gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact
     7681 tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccgataca
     7741 cctactcccc gattcgtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
     7801 tcaacttgaa aaagtggcac cgagtcggtg cttttttgtt ttagagctag aaatagcaag
     7861 ttaaaataag gctagtccgt ttttagcgcg tgcgccaatt ctgcagacaa atggctctag
     7921 aggtacccgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     7981 gcccattgac gtcaatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     8041 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     8101 ctattgacgt caatgacggt aaatggcccg cctggcattg tgcccagtac atgaccttat
     8161 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg
     8221 tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt
     8281 atttatttat tttttaatta ttttgtgcag cgatggggg
//
