LOCUS       sequence-273966-        9215 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gRNA to delete a nucleation site near HoxD. Use with HoxD del us1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9215)
  AUTHORS   Oksuz O, Narendra V, Lee CH, Descostes N, LeRoy G, Raviram R, 
            Blumenberg L, Karch K, Rocha PP, Garcia BA, Skok JA, Reinberg D
  TITLE     Capturing the Onset of PRC2-Mediated Repressive Domain Formation.
  JOURNAL   Mol Cell. 2018 Jun 21;70(6):1149-1162.e5. doi: 
            10.1016/j.molcel.2018.05.023.
  PUBMED    29932905
REFERENCE   2  (bases 1 to 9215)
  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.molcel.2018.05"; journalName: "Mol Cell"; date: 
            "2018-06-21- 21"; volume: "70"; issue: "6"; pages: "1149-1162.e5"
COMMENT     Sequence Label: HoxD del  ds1
FEATURES             Location/Qualifiers
     source          1..9215
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          93..320
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      332..341
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             341..406
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             413..433
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             458..4558
                     /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             458..4558
                     /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             4559..4606
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4622..4675
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(4676..4678,4679..5389)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4676..4678,4679..5389)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4979..4998)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5326..5347
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5417..5434)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5423..5630
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5639..5779
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5639..5768
                     /label=AAV2 ITR
     rep_origin      5854..6309
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5941..5960)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6151..6172
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6326..6345)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6445..6467
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6505..6523)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6591..6695
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6696..6764,6765..7556)
                     /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(6914..6933)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7727..8315
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8216..8235
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8377..8625
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8377..8397
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8548..8567
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8647..8722
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8819..9104
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc
       61 ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg
      121 tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc
      181 cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagctg agcaagaggt
      241 aagggtttaa gggatggttg gttggtgggg tattaatgtt taattacctg gagcacctgc
      301 ctgaaatcac tttttttcag gttggaccgg tgccaccatg gactataagg accacgacgg
      361 agactacaag gatcatgata ttgattacaa agacgatgac gataagatgg ccccaaagaa
      421 gaagcggaag gtcggtatcc acggagtccc agcagccgac aagaagtaca gcatcggcct
      481 ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag
      541 caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg
      601 agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag
      661 aagaagatac accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga
      721 gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga
      781 ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca
      841 cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc
      901 cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct
      961 gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt
     1021 gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa
     1081 ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct
     1141 gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc tgggcctgac
     1201 ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga
     1261 cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct
     1321 gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa
     1381 caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca
     1441 ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga
     1501 gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca
     1561 ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact
     1621 gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag
     1681 catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt
     1741 ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc
     1801 ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag
     1861 cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca
     1921 gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc
     1981 caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata
     2041 cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt
     2101 ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt
     2161 caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc
     2221 ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa
     2281 tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag
     2341 agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa
     2401 gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg
     2461 catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc
     2521 caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca
     2581 gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg
     2641 cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa
     2701 agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac
     2761 cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa
     2821 agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga
     2881 gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga
     2941 catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctgaagga
     3001 cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa
     3061 cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc
     3121 caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag
     3181 cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa
     3241 gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct
     3301 gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga
     3361 tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct
     3421 gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt
     3481 gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat
     3541 cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga
     3601 gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac
     3661 cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat
     3721 gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc
     3781 tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa
     3841 gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt
     3901 ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat
     3961 ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga
     4021 agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg
     4081 ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc
     4141 ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc
     4201 cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat
     4261 catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa
     4321 agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat
     4381 catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac
     4441 caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca
     4501 ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacaa
     4561 aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag aaaaaggaat tcggcagtgg
     4621 agagggcaga ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg gcccagtgag
     4681 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     4741 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     4801 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     4861 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     4921 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     4981 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     5041 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     5101 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     5161 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     5221 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     5281 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     5341 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg aattctaact
     5401 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     5461 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     5521 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     5581 caggacagca agggggagga ttgggaagag aatagcaggc atgctgggga gcggccgcag
     5641 gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc
     5701 gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga
     5761 gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     5821 atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg
     5881 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg
     5941 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     6001 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     6061 aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     6121 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     6181 tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt
     6241 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     6301 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     6361 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     6421 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     6481 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     6541 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     6601 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     6661 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     6721 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     6781 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     6841 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     6901 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     6961 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     7021 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     7081 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     7141 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     7201 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     7261 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     7321 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     7381 ttgctgataa atctggagcc ggtgagcgtg gaagccgcgg tatcattgca gcactggggc
     7441 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     7501 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     7561 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     7621 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     7681 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     7741 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     7801 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     7861 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     7921 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     7981 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     8041 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     8101 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     8161 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     8221 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     8281 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     8341 ggttcctggc cttttgctgg ccttttgctc acatgtgagg gcctatttcc catgattcct
     8401 tcatatttgc atatacgata caaggctgtt agagagataa ttggaattaa tttgactgta
     8461 aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt
     8521 gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat
     8581 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccggaca tctagcgcca
     8641 agcgcggttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
     8701 aaaagtggca ccgagtcggt gcttttttgt tttagagcta gaaatagcaa gttaaaataa
     8761 ggctagtccg tttttagcgc gtgcgccaat tctgcagaca aatggctcta gaggtacccg
     8821 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     8881 cgtcaatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     8941 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     9001 tcaatgacgg taaatggccc gcctggcatt gtgcccagta catgacctta tgggactttc
     9061 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca
     9121 cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg tatttattta
     9181 ttttttaatt attttgtgca gcgatggggg cgggg
//
