LOCUS       sequence-273960-        9521 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  gRNA to delete control region near HoxD. Use with HoxD del control 
            us.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9521)
  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 9521)
  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 control ds
FEATURES             Location/Qualifiers
     source          1..9521
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          215..442
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      454..463
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             463..528
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             535..555
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             580..4680
                     /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             580..4680
                     /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             4681..4728
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4744..4797
                     /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(4798..4800,4801..5511)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4798..4800,4801..5511)
                     /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(5101..5120)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5448..5469
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5539..5556)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5545..5752
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5761..5901
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5761..5890
                     /label=AAV2 ITR
     rep_origin      5976..6431
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6063..6082)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6273..6294
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6448..6467)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6567..6589
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6627..6645)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6713..6817
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6818..6886,6887..7678)
                     /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(7036..7055)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7849..8437
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8338..8357
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8499..8747
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8499..8519
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8670..8689
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8769..8844
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8941..9226
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     gap             9426..9484
                     /estimated_length=59
ORIGIN
        1 tkttkrtktt ktttktwatt tttkkggggg gggggggggg gggggggggg ggggggggcg
       61 cgcgcsrggg ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc
      121 ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg
      181 gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc
      241 cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact
      301 cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc tgagcaagag
      361 gtaagggttt aagggatggt tggttggtgg ggtattaatg tttaattacc tggagcacct
      421 gcctgaaatc actttttttc aggttggacc ggtgccacca tggactataa ggaccacgac
      481 ggagactaca aggatcatga tattgattac aaagacgatg acgataagat ggccccaaag
      541 aagaagcgga aggtcggtat ccacggagtc ccagcagccg acaagaagta cagcatcggc
      601 ctggacatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc
      661 agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc
      721 ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc
      781 agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac
      841 gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa
      901 gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac
      961 cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag
     1021 gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc
     1081 ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg
     1141 gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc
     1201 aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag
     1261 ctgcccggcg agaagaagaa tggcctgttc ggaaacctga ttgccctgag cctgggcctg
     1321 acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag
     1381 gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac
     1441 ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg
     1501 aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac
     1561 caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa
     1621 gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc
     1681 caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa
     1741 ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc
     1801 agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat
     1861 ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc
     1921 ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag
     1981 agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc
     2041 cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg
     2101 cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa
     2161 tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc
     2221 gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac
     2281 ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac
     2341 gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac
     2401 aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac
     2461 agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg
     2521 aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac
     2581 ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc
     2641 gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc
     2701 cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc
     2761 ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg
     2821 aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag
     2881 accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc
     2941 aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac
     3001 gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg
     3061 gacatcaacc ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag
     3121 gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac
     3181 aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac
     3241 gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg
     3301 agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca
     3361 aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag
     3421 ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag
     3481 gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac
     3541 ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc
     3601 gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa
     3661 atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc
     3721 gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa
     3781 accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc
     3841 atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag
     3901 tctatcctgc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct
     3961 aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa
     4021 gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc
     4081 atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa
     4141 gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac
     4201 ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg
     4261 ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc
     4321 cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag
     4381 atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac
     4441 aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat
     4501 atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac
     4561 accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc
     4621 caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac
     4681 aaaaggccgg cggccacgaa aaaggccggc caggcaaaaa agaaaaagga attcggcagt
     4741 ggagagggca gaggaagtct gctaacatgc ggtgacgtcg aggagaatcc tggcccagtg
     4801 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     4861 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
     4921 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     4981 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     5041 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
     5101 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     5161 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     5221 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     5281 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     5341 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     5401 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     5461 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa ggaattctaa
     5521 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     5581 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     5641 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     5701 ggcaggacag caagggggag gattgggaag agaatagcag gcatgctggg gagcggccgc
     5761 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     5821 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     5881 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     5941 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     6001 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     6061 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     6121 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     6181 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     6241 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     6301 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     6361 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     6421 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     6481 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     6541 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     6601 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     6661 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     6721 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6781 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6841 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6901 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     6961 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     7021 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     7081 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     7141 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     7201 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     7261 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     7321 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     7381 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     7441 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     7501 tattgctgat aaatctggag ccggtgagcg tggaagccgc ggtatcattg cagcactggg
     7561 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     7621 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     7681 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     7741 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     7801 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     7861 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     7921 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     7981 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     8041 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     8101 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     8161 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     8221 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     8281 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     8341 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     8401 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     8461 acggttcctg gccttttgct ggccttttgc tcacatgtga gggcctattt cccatgattc
     8521 cttcatattt gcatatacga tacaaggctg ttagagagat aattggaatt aatttgactg
     8581 taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt
     8641 ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt
     8701 atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccggc taatttagaa
     8761 cagcgcctgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
     8821 gaaaaagtgg caccgagtcg gtgctttttt gttttagagc tagaaatagc aagttaaaat
     8881 aaggctagtc cgtttttagc gcgtgcgcca attctgcaga caaatggctc tagaggtacc
     8941 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     9001 gacgtcaata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     9061 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     9121 cgtcaatgac ggtaaatggc ccgcctggca ttgtgcccag tacatgacct tatgggactt
     9181 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc
     9241 cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt
     9301 tattttttaa ttattttgtg cagcgatggg ggcggggggg ggggggggng cgcgccmggs
     9361 rnggggcggg gsgggggggg gggggggggg gggggggggg gggggggggg gggggggggg
     9421 gggggnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn
     9481 nnnngggggg gggggggggg gggggggsgs ggssrggggg g
//
