LOCUS       sequence-203440-        9261 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  CRISPR knockout. Expresses Cas9, EGFP, and sgRNA targeting PRKRA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9261)
  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 9261)
  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_PRKRA
FEATURES             Location/Qualifiers
     source          1..9261
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          133..360
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      372..381
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             381..446
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             453..473
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             498..4598
                     /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             498..4598
                     /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             4599..4646
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4662..4715
                     /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(4716..4718,4719..5429)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4716..4718,4719..5429)
                     /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(5019..5038)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5366..5387
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5457..5474)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5463..5670
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5679..5819
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5679..5808
                     /label=AAV2 ITR
     rep_origin      5894..6349
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5981..6000)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6191..6212
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6366..6385)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6485..6507
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6545..6563)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6631..6735
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6736..6804,6805..7596)
                     /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(6954..6973)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7767..8355
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8256..8275
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8417..8665
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8417..8437
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8588..8607
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8687..8762
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8859..9144
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg
       61 ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc
      121 gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc
      181 tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac
      241 ggcccttctc ctccgggctg taattagctg agcaagaggt aagggtttaa gggatggttg
      301 gttggtgggg tattaatgtt taattacctg gagcacctgc ctgaaatcac tttttttcag
      361 gttggaccgg tgccaccatg gactataagg accacgacgg agactacaag gatcatgata
      421 ttgattacaa agacgatgac gataagatgg ccccaaagaa gaagcggaag gtcggtatcc
      481 acggagtccc agcagccgac aagaagtaca gcatcggcct ggacatcggc accaactctg
      541 tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg
      601 gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg
      661 gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga
      721 agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca
      781 gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc
      841 accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct
      901 accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc
      961 tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc
     1021 ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt
     1081 tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac
     1141 tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg
     1201 gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact
     1261 tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg
     1321 acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc
     1381 tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc
     1441 ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga
     1501 aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca
     1561 agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca
     1621 tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag
     1681 aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc
     1741 tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca
     1801 accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg
     1861 ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct
     1921 ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga
     1981 ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg
     2041 agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa
     2101 agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca
     2161 accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg
     2221 actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg
     2281 atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc
     2341 tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc
     2401 tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat
     2461 acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg
     2521 gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc
     2581 tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc
     2641 agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg
     2701 gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc
     2761 ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga
     2821 acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc
     2881 tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc
     2941 tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact
     3001 acgatgtgga ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg
     3061 tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg
     3121 tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa
     3181 agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct
     3241 tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg
     3301 actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga
     3361 tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc
     3421 gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg
     3481 ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt
     3541 acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt
     3601 acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg
     3661 agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata
     3721 agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga
     3781 aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca
     3841 gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca
     3901 gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga
     3961 aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga
     4021 agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca
     4081 tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct
     4141 ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc
     4201 tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac
     4261 agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt
     4321 tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca
     4381 agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga
     4441 ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt
     4501 acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt
     4561 acgagacacg gatcgacctg tctcagctgg gaggcgacaa aaggccggcg gccacgaaaa
     4621 aggccggcca ggcaaaaaag aaaaaggaat tcggcagtgg agagggcaga ggaagtctgc
     4681 taacatgcgg tgacgtcgag gagaatcctg gcccagtgag caagggcgag gagctgttca
     4741 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     4801 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     4861 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     4921 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     4981 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     5041 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     5101 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     5161 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     5221 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     5281 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     5341 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     5401 tcactctcgg catggacgag ctgtacaagg aattctaact agagctcgct gatcagcctc
     5461 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     5521 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     5581 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     5641 ttgggaagag aatagcaggc atgctgggga gcggccgcag gaacccctag tgatggagtt
     5701 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     5761 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     5821 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     5881 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5941 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     6001 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     6061 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     6121 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     6181 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     6241 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     6301 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     6361 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     6421 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     6481 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     6541 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     6601 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     6661 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     6721 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     6781 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     6841 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     6901 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     6961 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     7021 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     7081 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     7141 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     7201 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     7261 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     7321 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     7381 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     7441 ggtgagcgtg gaagccgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     7501 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     7561 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     7621 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     7681 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     7741 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     7801 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     7861 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     7921 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     7981 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     8041 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     8101 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     8161 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     8221 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     8281 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     8341 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     8401 ccttttgctc acatgtgagg gcctatttcc catgattcct tcatatttgc atatacgata
     8461 caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca
     8521 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
     8581 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
     8641 tatcttgtgg aaaggacgaa acaccgagat gataacagct aagccagttt tagagctaga
     8701 aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
     8761 gcttttttgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg tttttagcgc
     8821 gtgcgccaat tctgcagaca aatggctcta gaggtacccg ttacataact tacggtaaat
     8881 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaatagt aacgccaata
     8941 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     9001 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     9061 gcctggcatt gtgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     9121 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
     9181 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
     9241 gcgatggggg cggggggggg g
//
