LOCUS       sequence-180045-       10675 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 10675)
  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 10675)
  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)_IRF8gRNA4
FEATURES             Location/Qualifiers
     source          1..10675
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          1..228
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             249..275
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             279..299
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(309..332,333..335,336..4409)
                     /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             309..4409
                     /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             4413..4433
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4470..4487)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4476..4683
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4692..4832
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4692..4821
                     /label=AAV2 ITR
     promoter        4977..5188
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             5201..5469
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      5498..5507
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5528..6190
                     /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             6260..6313
                     /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             6314..6913
                     /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(6314..6333)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6810..6830
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(6967..6987)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    7029..7150
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7066..7085)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7120..7139
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      7301..7756
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7388..7407)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7598..7619
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7773..7792)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7892..7914
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7952..7970)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8038..8142
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8143..8211,8212..9003)
                     /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(8361..8380)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9174..9762
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9663..9682
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        9824..10072
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     9824..9844
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     9995..10014
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        10093..10168
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        10265..10550
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
       61 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
      121 ccgggctgta attagctgag caagaggtaa gggtttaagg gatggttggt tggtggggta
      181 ttaatgttta attacctgga gcacctgcct gaaatcactt tttttcaggt tggaccggtg
      241 ccaccatgta cccatacgat gttccagatt acgcttcgcc gaagaaaaag cgcaaggtcg
      301 aagcgtccga caagaagtac agcatcggcc tggccatcgg caccaactct gtgggctggg
      361 ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg
      421 accggcacag catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag
      481 ccgaggccac ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga
      541 tctgctatct gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc
      601 acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct
      661 tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga
      721 gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg
      781 cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca
      841 gcgacgtgga caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa
      901 accccatcaa cgccagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga
      961 gcagacggct ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg
     1021 gcaacctgat tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg
     1081 ccgaggatgc caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc
     1141 tggcccagat cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg
     1201 ccatcctgct gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg
     1261 cctctatgat caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg
     1321 tgcggcagca gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct
     1381 acgccggcta cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca
     1441 tcctggaaaa gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc
     1501 tgcggaagca gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc
     1561 tgcacgccat tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa
     1621 agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa
     1681 acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg
     1741 aggaagtggt ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg
     1801 ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca
     1861 ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct
     1921 tcctgagcgg cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag
     1981 tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg
     2041 aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga
     2101 aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata
     2161 tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct
     2221 atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct
     2281 ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa
     2341 tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg
     2401 acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata
     2461 gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc
     2521 agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca
     2581 tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg
     2641 agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac
     2701 accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg
     2761 ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg
     2821 accatatcgt gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca
     2881 gaagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga
     2941 tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca
     3001 atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga
     3061 gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga
     3121 tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga
     3181 agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca
     3241 acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca
     3301 aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc
     3361 ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct
     3421 acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga
     3481 agcggcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg
     3541 attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg
     3601 aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc
     3661 tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg
     3721 tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga
     3781 gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca
     3841 tcgactttct ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc
     3901 ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg
     3961 aactgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg
     4021 ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg
     4081 tggaacagca caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga
     4141 gagtgatcct ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg
     4201 ataagcccat cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg
     4261 gagcccctgc cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca
     4321 ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac
     4381 ggatcgacct gtctcagctg ggaggcgaca gccccaagaa gaagagaaag gtggaggcca
     4441 gctaagaatt cctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     4501 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     4561 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     4621 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gagaatagca ggcatgctgg
     4681 ggagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     4741 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     4801 agtgagcgag cgagcgcgca gctgcctgca ggaaagagcc cacaacccct cacttggtgg
     4861 gccagtcctc tgatagactg tgtcccctgg atacccgtat tctagagcta gcgaattcga
     4921 atttaaatcg gatccgcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc
     4981 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg
     5041 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct
     5101 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt
     5161 tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc
     5221 acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc
     5281 tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc
     5341 gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac
     5401 gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg
     5461 ttacagatcc aagctgtgac cggcgcctac gctagacgcc accatggaga gcgacgagag
     5521 cggcctgccc gccatggaga tcgagtgccg catcaccggc accctgaacg gcgtggagtt
     5581 cgagctggtg ggcggcggag agggcacccc caagcagggc cgcatgacca acaagatgaa
     5641 gagcaccaaa ggcgccctga ccttcagccc ctacctgctg agccacgtga tgggctacgg
     5701 cttctaccac ttcggcacct accccagcgg ctacgagaac cccttcctgc acgccatcaa
     5761 caacggcggc tacaccaaca cccgcatcga gaagtacgag gacggcggcg tgctgcacgt
     5821 gagcttcagc taccgctacg aggccggccg cgtgatcggc gacttcaagg tggtgggcac
     5881 cggcttcccc gaggacagcg tgatcttcac cgacaagatc atccgcagca acgccaccgt
     5941 ggagcacctg caccccatgg gcgataacgt gctggtgggc agcttcgccc gcaccttcag
     6001 cctgcgcgac ggcggctact acagcttcgt ggtggacagc cacatgcact tcaagagcgc
     6061 catccacccc agcatcctgc agaacggggg ccccatgttc gccttccgcc gcgtggagga
     6121 gctgcacagc aacaccgagc tgggcatcgt ggagtaccag cacgccttca agacccccat
     6181 cgccttcgcc agatcccgcg ctcagtcgtc caattctgcc gtggacggca ccgccggacc
     6241 cggctccacc ggatctcgcg agggcagagg aagtcttcta acatgcggtg acgtggagga
     6301 gaatcccggc cctatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     6361 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     6421 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     6481 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     6541 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     6601 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     6661 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     6721 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     6781 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     6841 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     6901 gcccggtgcc tgaaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     6961 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     7021 gttaactaaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     7081 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     7141 atgtatctta tcatgtctgg aattgactca aatgatgtca attagtctat cagaagctca
     7201 tctggtctcc cttccgggcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     7261 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     7321 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     7381 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     7441 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     7501 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     7561 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     7621 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     7681 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     7741 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     7801 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     7861 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     7921 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     7981 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     8041 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     8101 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     8161 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     8221 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     8281 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     8341 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     8401 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     8461 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     8521 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     8581 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     8641 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     8701 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     8761 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     8821 tggtttattg ctgataaatc tggagccggt gagcgtggaa gccgcggtat cattgcagca
     8881 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     8941 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     9001 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     9061 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     9121 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     9181 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     9241 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     9301 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     9361 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     9421 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     9481 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     9541 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     9601 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     9661 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     9721 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     9781 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtgagggcc tatttcccat
     9841 gattccttca tatttgcata tacgatacaa ggctgttaga gagataattg gaattaattt
     9901 gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg
     9961 gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg
    10021 aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccgaataaga
    10081 gcccagattt tggttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca
    10141 acttgaaaaa gtggcaccga gtcggtgctt ttttgtttta gagctagaaa tagcaagtta
    10201 aaataaggct agtccgtttt tagcgcgtgc gccaattctg cagacaaatg gctctagagg
    10261 tacccgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
    10321 cattgacgtc aatagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
    10381 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
    10441 ttgacgtcaa tgacggtaaa tggcccgcct ggcattgtgc ccagtacatg accttatggg
    10501 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga
    10561 gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt
    10621 tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg gggcg
//
