LOCUS       sequence-180034-       10717 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Knockout IRF8 in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 10717)
  AUTHORS   Sontag S, Forster M, Qin J, Wanek P, Mitzka S, Schuler HM, 
            Koschmieder S, Rose-John S, Sere K, Zenke M
  TITLE     Modelling IRF8 Deficient Human Hematopoiesis and Dendritic Cell 
            Development with Engineered iPS Cells.
  JOURNAL   Stem Cells. 2017 Jan 16. doi: 10.1002/stem.2565.
  PUBMED    28090699
REFERENCE   2  (bases 1 to 10717)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Stem Cells.
            2017 Jan 16. doi: 10.1002/stem.2565."
COMMENT     Sequence Label: pX335A_hCas9(D10A)_IRF8gRNA3
FEATURES             Location/Qualifiers
     source          1..10717
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          3..230
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             251..277
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             281..301
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(311..334,335..337,338..4411)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             311..4411
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4415..4435
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4472..4489)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4478..4685
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4694..4834
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4694..4823
                     /label=AAV2 ITR
     promoter        4979..5190
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             5203..5471
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      5500..5509
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5530..6192
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
     CDS             6262..6315
                     /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             6316..6915
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6316..6335)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6812..6832
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6969..6989)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    7031..7152
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7068..7087)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7122..7141
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7303..7758
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7390..7409)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7600..7621
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7775..7794)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7894..7916
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7954..7972)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8040..8144
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8145..8213,8214..9005)
                     /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(8363..8382)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9176..9764
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9665..9684
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        9826..10074
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     9826..9846
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9997..10016
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        10095..10170
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        10267..10552
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
       61 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      121 ctccgggctg taattagctg agcaagaggt aagggtttaa gggatggttg gttggtgggg
      181 tattaatgtt taattacctg gagcacctgc ctgaaatcac tttttttcag gttggaccgg
      241 tgccaccatg tacccatacg atgttccaga ttacgcttcg ccgaagaaaa agcgcaaggt
      301 cgaagcgtcc gacaagaagt acagcatcgg cctggccatc ggcaccaact ctgtgggctg
      361 ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac
      421 cgaccggcac agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac
      481 agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg
      541 gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt
      601 ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat
      661 cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct
      721 gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct
      781 ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa
      841 cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga
      901 aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa
      961 gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt
     1021 cggcaacctg attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct
     1081 ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct
     1141 gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga
     1201 cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag
     1261 cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct
     1321 cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg
     1381 ctacgccggc tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc
     1441 catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct
     1501 gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga
     1561 gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga
     1621 aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg
     1681 aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt
     1741 cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt
     1801 cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt
     1861 caccgtgtat aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc
     1921 cttcctgagc ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa
     1981 agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt
     2041 ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct
     2101 gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga
     2161 tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac
     2221 ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg
     2281 ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac
     2341 aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca
     2401 cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga
     2461 tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct
     2521 gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa
     2581 catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg
     2641 cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga
     2701 acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa
     2761 tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt
     2821 ggaccatatc gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac
     2881 cagaagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa
     2941 gatgaagaac tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga
     3001 caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa
     3061 gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg
     3121 gatgaacact aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct
     3181 gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat
     3241 caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat
     3301 caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt
     3361 gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt
     3421 ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg
     3481 gaagcggcct ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg
     3541 ggattttgcc accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac
     3601 cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa
     3661 gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac
     3721 cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa
     3781 gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc
     3841 catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct
     3901 gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg
     3961 cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct
     4021 ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt
     4081 tgtggaacag cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa
     4141 gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg
     4201 ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct
     4261 gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag
     4321 caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac
     4381 acggatcgac ctgtctcagc tgggaggcga cagccccaag aagaagagaa aggtggaggc
     4441 cagctaagaa ttcctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     4501 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     4561 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     4621 gggggtgggg tggggcagga cagcaagggg gaggattggg aagagaatag caggcatgct
     4681 ggggagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     4741 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     4801 tcagtgagcg agcgagcgcg cagctgcctg caggaaagag cccacaaccc ctcacttggt
     4861 gggccagtcc tctgatagac tgtgtcccct ggatacccgt attctagagc tagcgaattc
     4921 gaatttaaat cggatccgcg gccgcgaagg atctgcgatc gctccggtgc ccgtcagtgg
     4981 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacg
     5041 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
     5101 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
     5161 tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc gaggggctcg catctctcct
     5221 tcacgcgccc gccgccctac ctgaggccgc catccacgcc ggttgagtcg cgttctgccg
     5281 cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt aaagctcagg
     5341 tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag ccggctctcc
     5401 acgctttgcc tgaccctgct tgctcaactc tacgtctttg tttcgttttc tgttctgcgc
     5461 cgttacagat ccaagctgtg accggcgcct acgctagacg ccaccatgga gagcgacgag
     5521 agcggcctgc ccgccatgga gatcgagtgc cgcatcaccg gcaccctgaa cggcgtggag
     5581 ttcgagctgg tgggcggcgg agagggcacc cccaagcagg gccgcatgac caacaagatg
     5641 aagagcacca aaggcgccct gaccttcagc ccctacctgc tgagccacgt gatgggctac
     5701 ggcttctacc acttcggcac ctaccccagc ggctacgaga accccttcct gcacgccatc
     5761 aacaacggcg gctacaccaa cacccgcatc gagaagtacg aggacggcgg cgtgctgcac
     5821 gtgagcttca gctaccgcta cgaggccggc cgcgtgatcg gcgacttcaa ggtggtgggc
     5881 accggcttcc ccgaggacag cgtgatcttc accgacaaga tcatccgcag caacgccacc
     5941 gtggagcacc tgcaccccat gggcgataac gtgctggtgg gcagcttcgc ccgcaccttc
     6001 agcctgcgcg acggcggcta ctacagcttc gtggtggaca gccacatgca cttcaagagc
     6061 gccatccacc ccagcatcct gcagaacggg ggccccatgt tcgccttccg ccgcgtggag
     6121 gagctgcaca gcaacaccga gctgggcatc gtggagtacc agcacgcctt caagaccccc
     6181 atcgccttcg ccagatcccg cgctcagtcg tccaattctg ccgtggacgg caccgccgga
     6241 cccggctcca ccggatctcg cgagggcaga ggaagtcttc taacatgcgg tgacgtggag
     6301 gagaatcccg gccctatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     6361 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     6421 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     6481 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     6541 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     6601 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     6661 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     6721 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     6781 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     6841 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     6901 aagcccggtg cctgaaatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     6961 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     7021 gcgttaacta aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     7081 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     7141 caatgtatct tatcatgtct ggaattgact caaatgatgt caattagtct atcagaagct
     7201 catctggtct cccttccggg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
     7261 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     7321 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct
     7381 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     7441 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     7501 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     7561 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     7621 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     7681 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     7741 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     7801 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     7861 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     7921 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     7981 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     8041 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     8101 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     8161 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     8221 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     8281 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     8341 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     8401 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     8461 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     8521 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     8581 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     8641 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     8701 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     8761 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     8821 gctggtttat tgctgataaa tctggagccg gtgagcgtgg aagccgcggt atcattgcag
     8881 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     8941 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     9001 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     9061 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     9121 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     9181 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     9241 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     9301 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     9361 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     9421 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     9481 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     9541 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     9601 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     9661 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     9721 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     9781 cctttttacg gttcctggcc ttttgctggc cttttgctca catgtgaggg cctatttccc
     9841 atgattcctt catatttgca tatacgatac aaggctgtta gagagataat tggaattaat
     9901 ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt
     9961 gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact
    10021 tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccgcgtaa
    10081 cctcgtcttc caaggtttta gagctagaaa tagcaagtta aaataaggct agtccgttat
    10141 caacttgaaa aagtggcacc gagtcggtgc ttttttgttt tagagctaga aatagcaagt
    10201 taaaataagg ctagtccgtt tttagcgcgt gcgccaattc tgcagacaaa tggctctaga
    10261 ggtacccgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
    10321 cccattgacg tcaatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
    10381 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
    10441 tattgacgtc aatgacggta aatggcccgc ctggcattgt gcccagtaca tgaccttatg
    10501 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt
    10561 gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta
    10621 tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggcgcgc
    10681 gccaggcggg gcggggcggg gcgaggggcg gggcggg
//
