LOCUS       sequence-203412-        9263 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  CRISPR knockout. Expresses Cas9, EGFP, and sgRNA targeting CELF1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9263)
  AUTHORS   Meyer C, Garzia A, Mazzola M, Gerstberger S, Molina H, Tuschl T
  TITLE     The TIA1 RNA-Binding Protein Family Regulates EIF2AK2-Mediated 
            Stress Response and Cell Cycle Progression.
  JOURNAL   Mol Cell. 2018 Feb 15;69(4):622-635.e6. doi: 
            10.1016/j.molcel.2018.01.011.
  PUBMED    29429924
REFERENCE   2  (bases 1 to 9263)
  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"; doi: 
            "10.1016/j.molcel.2018.01"; journalName: "Mol Cell"; date: 
            "2018-02-15- 15"; volume: "69"; issue: "4"; pages: "622-635.e6"
COMMENT     Sequence Label: px458_2A_GFP_sgRNA_CELF1
FEATURES             Location/Qualifiers
     source          1..9263
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          134..361
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      373..382
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             382..447
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             454..474
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             499..4599
                     /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             499..4599
                     /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             4600..4647
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4663..4716
                     /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(4717..4719,4720..5430)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4717..4719,4720..5430)
                     /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(5020..5039)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5367..5388
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5458..5475)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5464..5671
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5680..5820
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5680..5809
                     /label=AAV2 ITR
     rep_origin      5895..6350
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5982..6001)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6192..6213
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6367..6386)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6486..6508
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6546..6564)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6632..6736
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6737..6805,6806..7597)
                     /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(6955..6974)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7768..8356
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8257..8276
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8418..8666
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8418..8438
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8589..8608
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8688..8763
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8860..9145
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
       61 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
      121 cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc
      181 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
      241 cggcccttct cctccgggct gtaattagct gagcaagagg taagggttta agggatggtt
      301 ggttggtggg gtattaatgt ttaattacct ggagcacctg cctgaaatca ctttttttca
      361 ggttggaccg gtgccaccat ggactataag gaccacgacg gagactacaa ggatcatgat
      421 attgattaca aagacgatga cgataagatg gccccaaaga agaagcggaa ggtcggtatc
      481 cacggagtcc cagcagccga caagaagtac agcatcggcc tggacatcgg caccaactct
      541 gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt caaggtgctg
      601 ggcaacaccg accggcacag catcaagaag aacctgatcg gagccctgct gttcgacagc
      661 ggcgaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata caccagacgg
      721 aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa ggtggacgac
      781 agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg
      841 caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc
      901 taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat
      961 ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac
     1021 cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta caaccagctg
     1081 ttcgaggaaa accccatcaa cgccagcggc gtggacgcca aggccatcct gtctgccaga
     1141 ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga gaagaagaat
     1201 ggcctgttcg gaaacctgat tgccctgagc ctgggcctga cccccaactt caagagcaac
     1261 ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga cgacgacctg
     1321 gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc cgccaagaac
     1381 ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat caccaaggcc
     1441 cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct gaccctgctg
     1501 aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt cgaccagagc
     1561 aagaacggct acgccggcta cattgacggc ggagccagcc aggaagagtt ctacaagttc
     1621 atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa gctgaacaga
     1681 gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca ccagatccac
     1741 ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt cctgaaggac
     1801 aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg
     1861 gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc
     1921 tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat cgagcggatg
     1981 accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac
     2041 gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga
     2101 aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct gttcaagacc
     2161 aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc
     2221 gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac
     2281 gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt
     2341 ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg
     2401 ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga
     2461 tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc
     2521 ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag
     2581 ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc
     2641 cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag
     2701 ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag
     2761 cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag
     2821 aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc
     2881 ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac
     2941 ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac
     3001 tacgatgtgg accatatcgt gcctcagagc tttctgaagg acgactccat cgacaacaag
     3061 gtgctgacca gaagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc
     3121 gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga
     3181 aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc
     3241 ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg
     3301 gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg
     3361 atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg
     3421 cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc
     3481 gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg
     3541 tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag
     3601 tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc
     3661 gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat
     3721 aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg
     3781 aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac
     3841 agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgac
     3901 agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag
     3961 aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag
     4021 aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc
     4081 atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc
     4141 tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc
     4201 ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa
     4261 cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca gatcagcgag
     4321 ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc cgcctacaac
     4381 aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct gtttaccctg
     4441 accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga ccggaagagg
     4501 tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg
     4561 tacgagacac ggatcgacct gtctcagctg ggaggcgaca aaaggccggc ggccacgaaa
     4621 aaggccggcc aggcaaaaaa gaaaaaggaa ttcggcagtg gagagggcag aggaagtctg
     4681 ctaacatgcg gtgacgtcga ggagaatcct ggcccagtga gcaagggcga ggagctgttc
     4741 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     4801 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     4861 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     4921 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     4981 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     5041 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     5101 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     5161 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     5221 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     5281 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     5341 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     5401 atcactctcg gcatggacga gctgtacaag gaattctaac tagagctcgc tgatcagcct
     5461 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     5521 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     5581 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     5641 attgggaaga gaatagcagg catgctgggg agcggccgca ggaaccccta gtgatggagt
     5701 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     5761 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     5821 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     5881 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     5941 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     6001 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     6061 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     6121 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     6181 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     6241 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     6301 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     6361 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     6421 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     6481 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     6541 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     6601 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     6661 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     6721 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     6781 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     6841 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     6901 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     6961 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     7021 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     7081 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     7141 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     7201 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     7261 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     7321 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     7381 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     7441 cggtgagcgt ggaagccgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     7501 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     7561 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     7621 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     7681 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     7741 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     7801 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     7861 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     7921 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     7981 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     8041 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     8101 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     8161 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     8221 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     8281 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     8341 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     8401 gccttttgct cacatgtgag ggcctatttc ccatgattcc ttcatatttg catatacgat
     8461 acaaggctgt tagagagata attggaatta atttgactgt aaacacaaag atattagtac
     8521 aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt
     8581 ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat
     8641 atatcttgtg gaaaggacga aacaccgcat ggcacagacg gctatcagtt ttagagctag
     8701 aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg
     8761 tgcttttttg ttttagagct agaaatagca agttaaaata aggctagtcc gtttttagcg
     8821 cgtgcgccaa ttctgcagac aaatggctct agaggtaccc gttacataac ttacggtaaa
     8881 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaatag taacgccaat
     8941 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     9001 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     9061 cgcctggcat tgtgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     9121 cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc
     9181 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
     9241 agcgatgggg gcgggggggg ggg
//
