LOCUS       sequence-458440-        9290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  single guide RNA targeting human RB1 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9290)
  AUTHORS   Lindstrom L, Zhou J, Villoutreix BO, Malycheva D, Otrocka M, 
            Gustavsson AL, Lundback T, Bliman D, Shameem MA, Straw M, Riesbeck 
            K, Olsson R, Alvarado-Kristensson M
  TITLE     Targeting TUBG1 in RB1-negative tumors.
  JOURNAL   FASEB J. 2025 Mar 15;39(5):e70431. doi: 10.1096/fj.202403180RR.
  PUBMED    40019206
REFERENCE   2  (bases 1 to 9290)
  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: "FASEB J."; 
            date: "2025-03-15"; volume: "39(5):e70431. doi"; pages: " 
            10.1096/fj.202403180RR"
COMMENT     Sequence Label: pSpCas9 (BB)-2A-GFP-RB1sg
FEATURES             Location/Qualifiers
     source          1..9290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             192..239
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             255..308
                     /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(309..311,312..1022)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(309..311,312..1022)
                     /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(612..631)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     959..980
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1050..1067)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1056..1263
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   1272..1412
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1272..1401
                     /label=AAV2 ITR
     rep_origin      1487..1942
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1574..1593)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1784..1805
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1959..1978)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2078..2100
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2138..2156)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2224..2328
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2329..2397,2398..3189)
                     /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(2547..2566)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3360..3948
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3849..3868
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4010..4258
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     4010..4030
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     4181..4200
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        4280..4355
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        4452..4737
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        4739..5015
                     /label=chicken beta-actin promoter
     intron          5016..5243
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      5255..5264
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5264..5329
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5336..5356
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(5381..9290,1..191)
                     /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             join(5381..9290,1..191)
                     /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"
ORIGIN
        1 aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca
       61 agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg
      121 ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc
      181 tgggaggcga caaaaggccg gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagg
      241 aattcggcag tggagagggc agaggaagtc tgctaacatg cggtgacgtc gaggagaatc
      301 ctggcccagt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
      361 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
      421 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
      481 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
      541 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
      601 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
      661 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
      721 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
      781 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
      841 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
      901 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
      961 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1021 aggaattcta actagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     1081 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     1141 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     1201 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gagaatagca ggcatgctgg
     1261 ggagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     1321 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     1381 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     1441 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     1501 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     1561 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     1621 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     1681 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     1741 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     1801 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
     1861 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     1921 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     1981 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     2041 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     2101 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     2161 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     2221 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2281 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2341 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2401 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2461 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2521 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     2581 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2641 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2701 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     2761 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     2821 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     2881 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     2941 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3001 gctggctggt ttattgctga taaatctgga gccggtgagc gtggaagccg cggtatcatt
     3061 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3121 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3181 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3241 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3301 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3361 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3421 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3481 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     3541 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3601 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3661 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3721 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3781 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3841 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3901 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3961 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtg agggcctatt
     4021 tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat
     4081 taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt
     4141 tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt
     4201 aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgc
     4261 ggtggcggcc gtttttcggg ttttagagct agaaatagca agttaaaata aggctagtcc
     4321 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgttttagag ctagaaatag
     4381 caagttaaaa taaggctagt ccgtttttag cgcgtgcgcc aattctgcag acaaatggct
     4441 ctagaggtac ccgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     4501 ccccgcccat tgacgtcaat agtaacgcca atagggactt tccattgacg tcaatgggtg
     4561 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     4621 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attgtgccca gtacatgacc
     4681 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc
     4741 gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt
     4801 ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg gggggggggc
     4861 gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg
     4921 cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc
     4981 ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc
     5041 ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac
     5101 tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag ctgagcaaga
     5161 ggtaagggtt taagggatgg ttggttggtg gggtattaat gtttaattac ctggagcacc
     5221 tgcctgaaat cacttttttt caggttggac cggtgccacc atggactata aggaccacga
     5281 cggagactac aaggatcatg atattgatta caaagacgat gacgataaga tggccccaaa
     5341 gaagaagcgg aaggtcggta tccacggagt cccagcagcc gacaagaagt acagcatcgg
     5401 cctggacatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc
     5461 cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat
     5521 cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga agagaaccgc
     5581 cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa
     5641 cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga
     5701 agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta
     5761 ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa
     5821 ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt
     5881 cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct
     5941 ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc
     6001 caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca
     6061 gctgcccggc gagaagaaga atggcctgtt cggaaacctg attgccctga gcctgggcct
     6121 gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa
     6181 ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga
     6241 cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt
     6301 gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca
     6361 ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa
     6421 agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag
     6481 ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga
     6541 actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg
     6601 cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga
     6661 tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat
     6721 cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa
     6781 gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc
     6841 ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct
     6901 gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa
     6961 atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat
     7021 cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta
     7081 cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa
     7141 cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga
     7201 caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga
     7261 cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat
     7321 gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa
     7381 cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt
     7441 cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat
     7501 ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc
     7561 cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt
     7621 gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca
     7681 gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat
     7741 caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa
     7801 cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact
     7861 ggacatcaac cggctgtccg actacgatgt ggaccatatc gtgcctcaga gctttctgaa
     7921 ggacgactcc atcgacaaca aggtgctgac cagaagcgac aagaaccggg gcaagagcga
     7981 caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcggc agctgctgaa
     8041 cgccaagctg attacccaga gaaagttcga caatctgacc aaggccgaga gaggcggcct
     8101 gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac
     8161 aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg agaatgacaa
     8221 gctgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa
     8281 ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta
     8341 cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg aaagcgagtt
     8401 cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga
     8461 aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac
     8521 cgagattacc ctggccaacg gcgagatccg gaagcggcct ctgatcgaga caaacggcga
     8581 aaccggggag atcgtgtggg ataagggccg ggattttgcc accgtgcgga aagtgctgag
     8641 catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga
     8701 gtctatcctg cccaagagga acagcgataa gctgatcgcc agaaagaagg actgggaccc
     8761 taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg tggtggccaa
     8821 agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat
     8881 catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa
     8941 agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa
     9001 cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg aactggccct
     9061 gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc
     9121 ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaagcact acctggacga
     9181 gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga
     9241 caaagtgctg tccgcctaca acaagcaccg ggataagccc atcagagagc
//
