LOCUS       sequence-273958-        9230 bp    DNA     circular SYN 15-APR-2025
DEFINITION  gRNA  to delete a nucleation site near HoxD. Use with HoxD del ds1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9230)
  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 9230)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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  us1
FEATURES             Location/Qualifiers
     source          1..9230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          105..332
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      344..353
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             353..418
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             425..445
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             470..4570
                     /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             470..4570
                     /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             4571..4618
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4634..4687
                     /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(4688..4690,4691..5401)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4688..4690,4691..5401)
                     /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(4991..5010)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5338..5359
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5429..5446)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5435..5642
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5651..5791
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5651..5780
                     /label=AAV2 ITR
     rep_origin      5866..6321
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5953..5972)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6163..6184
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6338..6357)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6457..6479
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6517..6535)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6603..6707
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6708..6776,6777..7568)
                     /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(6926..6945)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7739..8327
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8228..8247
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8389..8637
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8389..8409
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8560..8579
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8659..8734
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8831..9116
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
       61 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct
      121 gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga
      181 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
      241 tgagcaagag gtaagggttt aagggatggt tggttggtgg ggtattaatg tttaattacc
      301 tggagcacct gcctgaaatc actttttttc aggttggacc ggtgccacca tggactataa
      361 ggaccacgac ggagactaca aggatcatga tattgattac aaagacgatg acgataagat
      421 ggccccaaag aagaagcgga aggtcggtat ccacggagtc ccagcagccg acaagaagta
      481 cagcatcggc ctggacatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta
      541 caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa
      601 gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa
      661 gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat
      721 cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt
      781 cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga
      841 ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag
      901 caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg
      961 gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt
     1021 catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg
     1081 cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct
     1141 gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggaaacctga ttgccctgag
     1201 cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca
     1261 gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca
     1321 gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat
     1381 cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata
     1441 cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga
     1501 gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg
     1561 cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg
     1621 caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt
     1681 cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg
     1741 gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac
     1801 cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat
     1861 gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg
     1921 cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga
     1981 gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac
     2041 caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa
     2101 aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa
     2161 agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga
     2221 tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga
     2281 cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact
     2341 gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga
     2401 caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa
     2461 gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc
     2521 cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa
     2581 agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc
     2641 caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga
     2701 cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag
     2761 agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga
     2821 agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca
     2881 gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga
     2941 ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gaccatatcg tgcctcagag
     3001 ctttctgaag gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg
     3061 caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca
     3121 gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag
     3181 aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg
     3241 gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga
     3301 gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga
     3361 tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca
     3421 cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga
     3481 aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag
     3541 cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt
     3601 tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac
     3661 aaacggcgaa accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa
     3721 agtgctgagc atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt
     3781 cagcaaagag tctatcctgc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga
     3841 ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt
     3901 ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg
     3961 gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa
     4021 gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga
     4081 gctggaaaac ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga
     4141 actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct
     4201 gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta
     4261 cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc
     4321 taatctggac aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca
     4381 ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa
     4441 gtactttgac accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc
     4501 caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct
     4561 gggaggcgac aaaaggccgg cggccacgaa aaaggccggc caggcaaaaa agaaaaagga
     4621 attcggcagt ggagagggca gaggaagtct gctaacatgc ggtgacgtcg aggagaatcc
     4681 tggcccagtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     4741 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     4801 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     4861 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     4921 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     4981 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     5041 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     5101 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     5161 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     5221 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     5281 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     5341 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     5401 ggaattctaa ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     5461 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     5521 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     5581 ggtggggtgg ggcaggacag caagggggag gattgggaag agaatagcag gcatgctggg
     5641 gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg
     5701 ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca
     5761 gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg
     5821 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg
     5881 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     5941 ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     6001 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     6061 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     6121 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     6181 ctggaacaac actcaactct atctcgggct attcttttga tttataaggg attttgccga
     6241 tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     6301 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     6361 agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     6421 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     6481 tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat
     6541 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     6601 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     6661 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     6721 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     6781 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     6841 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     6901 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     6961 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     7021 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     7081 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     7141 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     7201 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     7261 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     7321 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     7381 ctggctggtt tattgctgat aaatctggag ccggtgagcg tggaagccgc ggtatcattg
     7441 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     7501 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     7561 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     7621 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     7681 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     7741 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     7801 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     7861 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     7921 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     7981 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     8041 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     8101 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     8161 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     8221 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     8281 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     8341 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtga gggcctattt
     8401 cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat aattggaatt
     8461 aatttgactg taaacacaaa gatattagta caaaatacgt gacgtagaaa gtaataattt
     8521 cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg actatcatat gcttaccgta
     8581 acttgaaagt atttcgattt cttggcttta tatatcttgt ggaaaggacg aaacaccggt
     8641 tgtggctgat cagcctcggt tttagagcta gaaatagcaa gttaaaataa ggctagtccg
     8701 ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt gttttagagc tagaaatagc
     8761 aagttaaaat aaggctagtc cgtttttagc gcgtgcgcca attctgcaga caaatggctc
     8821 tagaggtacc cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     8881 cccgcccatt gacgtcaata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     8941 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     9001 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttgtgcccag tacatgacct
     9061 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg
     9121 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
     9181 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg
//
