LOCUS       sequence-175664-        8328 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 8328)
  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 8328)
  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-2
FEATURES             Location/Qualifiers
     source          1..8328
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      223..232
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             232..297
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             304..324
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(349..372,373..375,376..4449)
                     /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             349..4449
                     /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             4450..4497
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4525..4542)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4531..4738
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4747..4887
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4747..4876
                     /label=AAV2 ITR
     rep_origin      4962..5417
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5049..5068)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5259..5280
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5434..5453)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5553..5575
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5613..5631)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5699..5803
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5804..5872,5873..6664)
                     /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(6022..6041)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6835..7423
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7324..7343
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7485..7733
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7485..7505
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7656..7675
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7754..7829
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        7926..8211
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac
       61 cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagct
      121 gagcaagagg taagggttta agggatggtt ggttggtggg gtattaatgt ttaattacct
      181 ggagcacctg cctgaaatca ctttttttca ggttggaccg gtgccaccat ggactataag
      241 gaccacgacg gagactacaa ggatcatgat attgattaca aagacgatga cgataagatg
      301 gccccaaaga agaagcggaa ggtcggtatc cacggagtcc cagcagccga caagaagtac
      361 agcatcggcc tggccatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac
      421 aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag
      481 aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag
      541 agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc
      601 ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc
      661 ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag
      721 gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc
      781 accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg
      841 ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc
      901 atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc
      961 gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg
     1021 atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc
     1081 ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag
     1141 ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag
     1201 tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc
     1261 ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac
     1321 gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag
     1381 aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc
     1441 ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc
     1501 accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc
     1561 gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg
     1621 caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc
     1681 ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg
     1741 accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc
     1801 gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag
     1861 aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc
     1921 aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa
     1981 aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa
     2041 gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat
     2101 cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac
     2161 ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg
     2221 tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac
     2281 aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag
     2341 ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc
     2401 gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa
     2461 gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc
     2521 aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac
     2581 gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga
     2641 gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa
     2701 gagggcatca aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag
     2761 ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac
     2821 caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc
     2881 tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc
     2941 aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag
     3001 ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga
     3061 ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg
     3121 cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag
     3181 aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat
     3241 ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac
     3301 gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa
     3361 agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc
     3421 gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt
     3481 ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca
     3541 aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa
     3601 gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc
     3661 agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac
     3721 tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg
     3781 gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg
     3841 atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag
     3901 ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag
     3961 ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa
     4021 ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg
     4081 aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac
     4141 ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct
     4201 aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag
     4261 gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag
     4321 tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc
     4381 accctgatcc accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg
     4441 ggaggcgaca aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa gaaaaagtaa
     4501 gaattcctag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     4561 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     4621 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     4681 gggtggggca ggacagcaag ggggaggatt gggaagagaa tagcaggcat gctggggagc
     4741 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     4801 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     4861 gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc
     4921 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     4981 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt
     5041 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     5101 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     5161 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     5221 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     5281 aacaacactc aactctatct cgggctattc ttttgattta taagggattt tgccgatttc
     5341 ggtctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     5401 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     5461 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     5521 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     5581 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     5641 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     5701 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5761 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     5821 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     5881 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     5941 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     6001 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     6061 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     6121 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     6181 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     6241 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     6301 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     6361 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     6421 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     6481 ctggtttatt gctgataaat ctggagccgg tgagcgtgga agccgcggta tcattgcagc
     6541 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6601 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     6661 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     6721 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6781 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6841 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6901 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6961 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     7021 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     7081 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7141 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     7201 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     7261 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     7321 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     7381 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7441 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtgagggc ctatttccca
     7501 tgattccttc atatttgcat atacgataca aggctgttag agagataatt ggaattaatt
     7561 tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg
     7621 ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt
     7681 gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgaaac accgaggctg
     7741 ttgatcgcgc ggcgttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
     7801 aacttgaaaa agtggcaccg agtcggtgct tttttgtttt agagctagaa atagcaagtt
     7861 aaaataaggc tagtccgttt ttagcgcgtg cgccaattct gcagacaaat ggctctagag
     7921 gtacccgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     7981 ccattgacgt caatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     8041 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     8101 attgacgtca atgacggtaa atggcccgcc tggcattgtg cccagtacat gaccttatgg
     8161 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg
     8221 agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat
     8281 ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggg
//
