LOCUS       sequence-182382-        9128 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  expresses Cas9 and gRNA targeting PAFAH1B1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9128)
  AUTHORS   Josipovic I, Fork C, Preussner J, Prior KK, Iloska D, Vasconez AE, 
            Labocha S, Angioni C, Thomas D, Ferreiros N, Looso M, Pullamsetti 
            SS, Geisslinger G, Steinhilber D, Brandes RP, Leisegang MS
  TITLE     PAFAH1B1 and the lncRNA NONHSAT073641 maintain an angiogenic 
            phenotype in human endothelial cells.
  JOURNAL   Acta Physiol (Oxf). 2016 Apr 28. doi: 10.1111/apha.12700.
  PUBMED    27124368
REFERENCE   2  (bases 1 to 9128)
  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: "Acta 
            Physiol (Oxf). 2016 Apr 28. doi: 10.1111/apha.12700."
COMMENT     Sequence Label: pSpCas9(BB)-2A-Puro-Exon4-PAFAH1B1
FEATURES             Location/Qualifiers
     source          1..9128
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          94..321
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      333..342
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             342..407
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             414..434
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             459..4559
                     /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             459..4559
                     /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             4560..4607
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4623..4676
                     /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             4677..5276
                     /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(4677..4696)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5173..5193
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5304..5321)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5310..5517
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5526..5666
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5526..5655
                     /label=AAV2 ITR
     rep_origin      5741..6196
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5828..5847)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6038..6059
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6213..6232)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6332..6354
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6392..6410)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6478..6582
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6583..6651,6652..7443)
                     /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(6801..6820)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7614..8202
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8103..8122
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8264..8512
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8264..8284
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8435..8454
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8534..8609
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8706..8991
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
       61 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc
      121 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
      181 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagct gagcaagagg
      241 taagggttta agggatggtt ggttggtggg gtattaatgt ttaattacct ggagcacctg
      301 cctgaaatca ctttttttca ggttggaccg gtgccaccat ggactataag gaccacgacg
      361 gagactacaa ggatcatgat attgattaca aagacgatga cgataagatg gccccaaaga
      421 agaagcggaa ggtcggtatc cacggagtcc cagcagccga caagaagtac agcatcggcc
      481 tggacatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca
      541 gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg
      601 gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca
      661 gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg
      721 agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag
      781 aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc
      841 acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg
      901 ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc
      961 tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg
     1021 tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca
     1081 aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc
     1141 tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga
     1201 cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg
     1261 acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc
     1321 tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga
     1381 acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc
     1441 accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag
     1501 agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc
     1561 aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac
     1621 tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca
     1681 gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt
     1741 tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc
     1801 cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga
     1861 gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc
     1921 agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc
     1981 ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat
     2041 acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg
     2101 tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact
     2161 tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg
     2221 cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca
     2281 atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca
     2341 gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga
     2401 agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg
     2461 gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg
     2521 ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc
     2581 agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg
     2641 gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga
     2701 aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga
     2761 ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca
     2821 aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg
     2881 agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg
     2941 acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc tttctgaagg
     3001 acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca
     3061 acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg
     3121 ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga
     3181 gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa
     3241 agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc
     3301 tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg
     3361 atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc
     3421 tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg
     3481 tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa
     3541 tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg
     3601 agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa
     3661 ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca
     3721 tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt
     3781 ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta
     3841 agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag
     3901 tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca
     3961 tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag
     4021 aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg
     4081 gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc
     4141 cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc
     4201 ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga
     4261 tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca
     4321 aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata
     4381 tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca
     4441 ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc
     4501 accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca
     4561 aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa gaaaaaggaa ttcggcagtg
     4621 gagagggcag aggaagtctg ctaacatgcg gtgacgtcga ggagaatcct ggcccaatga
     4681 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca
     4741 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc
     4801 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     4861 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     4921 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     4981 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     5041 cgtggttcct ggccaccgtc ggagtctcgc ccgaccacca gggcaagggt ctgggcagcg
     5101 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     5161 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     5221 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgagaat
     5281 tctaactaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     5341 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     5401 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     5461 ggtggggcag gacagcaagg gggaggattg ggaagagaat agcaggcatg ctggggagcg
     5521 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     5581 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     5641 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     5701 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     5761 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     5821 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     5881 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     5941 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     6001 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     6061 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     6121 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     6181 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     6241 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     6301 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     6361 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     6421 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     6481 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     6541 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     6601 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     6661 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     6721 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     6781 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     6841 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     6901 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     6961 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     7021 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     7081 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     7141 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     7201 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     7261 tggtttattg ctgataaatc tggagccggt gagcgtggaa gccgcggtat cattgcagca
     7321 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     7381 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     7441 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     7501 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     7561 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     7621 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     7681 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     7741 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     7801 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     7861 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     7921 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     7981 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     8041 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     8101 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     8161 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     8221 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtgagggcc tatttcccat
     8281 gattccttca tatttgcata tacgatacaa ggctgttaga gagataattg gaattaattt
     8341 gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg
     8401 gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg
     8461 aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccggcatatt
     8521 tttctggcgg acggttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
     8581 aacttgaaaa agtggcaccg agtcggtgct tttttgtttt agagctagaa atagcaagtt
     8641 aaaataaggc tagtccgttt ttagcgcgtg cgccaattct gcagacaaat ggctctagag
     8701 gtacccgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     8761 ccattgacgt caatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     8821 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     8881 attgacgtca atgacggtaa atggcccgcc tggcattgtg cccagtacat gaccttatgg
     8941 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg
     9001 agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat
     9061 ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggggcgcgcg
     9121 ccaggcgg
//
