LOCUS       sequence-120925-        8509 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  specific sgRNA against mouse Dicer1 gene cloned in the pX330 
            backbone (Addgene Number 42230). Generation of Dicer1 knockout 
            mESCs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8509)
  AUTHORS   Bodak M, Cirera-Salinas D, Yu J, Ngondo RP, Ciaudo C
  TITLE     Dicer, a new regulator of pluripotency exit and LINE-1 elements in 
            mouse embryonic stem cells.
  JOURNAL   FEBS Open Bio. 2017 Jan 11;7(2):204-220. doi: 
            10.1002/2211-5463.12174. eCollection 2017 Feb.
  PUBMED    28174687
REFERENCE   2  (bases 1 to 8509)
  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: 
            "10.1002/2211-5463.12174"; journalName: "FEBS Open Bio"; date: 
            "2017-01-11- 11"; volume: "7"; issue: "2"; pages: "204-220"
COMMENT     Sequence Label: pX330-sgRNA_Dicer_1
FEATURES             Location/Qualifiers
     source          1..8509
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          45..273
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      285..294
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             294..359
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             366..386
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             411..4511
                     /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             411..4511
                     /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             4512..4559
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(4587..4604)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4593..4800
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4809..4949
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4809..4938
                     /label=AAV2 ITR
     rep_origin      5024..5479
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5111..5130)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5321..5342
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5496..5515)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5615..5637
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5675..5693)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5761..5865
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5866..5934,5935..6726)
                     /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(6084..6103)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6897..7485
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7386..7405
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        7547..7795
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     7547..7567
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7718..7737
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        7817..7892
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        7989..8274
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        join(8276..8509,1..44)
                     /label=chicken beta-actin promoter
ORIGIN
        1 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgacgc
       61 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
      121 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
      181 ctgagcaaga ggtaagggtt taagggatgg ttggttggtg gggtattaat gtttaattac
      241 ctggagcacc tgcctgaaat cacttttttt caggttggac cggtgccacc atggactata
      301 aggaccacga cggagactac aaggatcatg atattgatta caaagacgat gacgataaga
      361 tggccccaaa gaagaagcgg aaggtcggta tccacggagt cccagcagcc gacaagaagt
      421 acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt
      481 acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga
      541 agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga
      601 agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga
      661 tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct
      721 tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg
      781 aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca
      841 gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc
      901 ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt
      961 tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg
     1021 gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc
     1081 tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg attgccctga
     1141 gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc
     1201 agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc
     1261 agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca
     1321 tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat
     1381 acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg
     1441 agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg
     1501 gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg
     1561 gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct
     1621 tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc
     1681 ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga
     1741 ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga
     1801 tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg
     1861 gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg
     1921 agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga
     1981 ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga
     2041 aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga
     2101 aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag
     2161 atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg
     2221 acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac
     2281 tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg
     2341 acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga
     2401 agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt
     2461 ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta
     2521 aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg
     2581 ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg
     2641 acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca
     2701 gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg
     2761 aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc
     2821 agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg
     2881 accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc gtgcctcaga
     2941 gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac aagaaccggg
     3001 gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcggc
     3061 agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc aaggccgaga
     3121 gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc
     3181 ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg
     3241 agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg
     3301 atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc
     3361 acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg
     3421 aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga
     3481 gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact
     3541 ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct ctgatcgaga
     3601 caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc accgtgcgga
     3661 aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct
     3721 tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc agaaagaagg
     3781 actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg
     3841 tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg
     3901 ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca
     3961 agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg
     4021 agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg
     4081 aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc
     4141 tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaagcact
     4201 acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg
     4261 ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc atcagagagc
     4321 aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca
     4381 agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg
     4441 ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc
     4501 tgggaggcga caaaaggccg gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagt
     4561 aagaattcct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     4621 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     4681 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     4741 tggggtgggg caggacagca agggggagga ttgggaagag aatagcaggc atgctgggga
     4801 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     4861 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     4921 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
     4981 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     5041 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     5101 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     5161 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     5221 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     5281 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     5341 ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     5401 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     5461 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     5521 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     5581 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     5641 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     5701 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     5761 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5821 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5881 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5941 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     6001 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     6061 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     6121 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     6181 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     6241 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     6301 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6361 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6421 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6481 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6541 ggctggttta ttgctgataa atctggagcc ggtgagcgtg gaagccgcgg tatcattgca
     6601 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6661 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6721 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6781 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6841 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6901 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6961 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7021 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     7081 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7141 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7201 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7261 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7321 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7381 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7441 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7501 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtgagg gcctatttcc
     7561 catgattcct tcatatttgc atatacgata caaggctgtt agagagataa ttggaattaa
     7621 tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct
     7681 tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac
     7741 ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccgggag
     7801 tttaaaccta caggcggttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
     7861 atcaacttga aaaagtggca ccgagtcggt gcttttttgt tttagagcta gaaatagcaa
     7921 gttaaaataa ggctagtccg tttttagcgc gtgcgccaat tctgcagaca aatggctcta
     7981 gaggtacccg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     8041 cgcccattga cgtcaatagt aacgccaata gggactttcc attgacgtca atgggtggag
     8101 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     8161 cctattgacg tcaatgacgg taaatggccc gcctggcatt gtgcccagta catgacctta
     8221 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag
     8281 gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg
     8341 tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc
     8401 gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg
     8461 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggc
//
