LOCUS       sequence-171515-        8993 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses sgRNA targeting human TP53 around codon R273.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8993)
  AUTHORS   Guernet A, Mungamuri SK, Cartier D, Sachidanandam R, Jayaprakash A, 
            Adriouch S, Vezain M, Charbonnier F, Rohkin G, Coutant S, Yao S, 
            Ainani H, Alexandre D, Tournier I, Boyer O, Aaronson SA, Anouar Y, 
            Grumolato L
  TITLE     CRISPR-Barcoding for Intratumor Genetic Heterogeneity Modeling and 
            Functional Analysis of Oncogenic Driver Mutations.
  JOURNAL   Mol Cell. 2016 Aug 4;63(3):526-38. doi: 
            10.1016/j.molcel.2016.06.017. Epub 2016 Jul 21.
  PUBMED    27453044
REFERENCE   2  (bases 1 to 8993)
  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.2016.06.017"; journalName: "Mol Cell"; date: 
            "2016-08-4- 4"; volume: "63"; issue: "3"; pages: "526-38"
COMMENT     Sequence Label: sgTP53(R273)
FEATURES             Location/Qualifiers
     source          1..8993
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          1..228
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      240..249
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             249..314
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             321..341
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             366..4466
                     /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             366..4466
                     /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             4467..4514
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4530..4583
                     /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             4584..5183
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4584..4603)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     complement(5211..5228)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5217..5424
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5433..5573
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5433..5562
                     /label=AAV2 ITR
     rep_origin      5648..6103
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5735..5754)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5945..5966
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6120..6139)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6239..6261
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6299..6317)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6385..6489
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6490..6558,6559..7350)
                     /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(6708..6727)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7521..8109
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8010..8029
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8171..8419
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8171..8191
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8342..8361
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8441..8516
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8613..8898
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
       61 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
      121 ccgggctgta attagctgag caagaggtaa gggtttaagg gatggttggt tggtggggta
      181 ttaatgttta attacctgga gcacctgcct gaaatcactt tttttcaggt tggaccggtg
      241 ccaccatgga ctataaggac cacgacggag actacaagga tcatgatatt gattacaaag
      301 acgatgacga taagatggcc ccaaagaaga agcggaaggt cggtatccac ggagtcccag
      361 cagccgacaa gaagtacagc atcggcctgg acatcggcac caactctgtg ggctgggccg
      421 tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc
      481 ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg
      541 aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct
      601 gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca
      661 gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg
      721 gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa
      781 agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc
      841 acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg
      901 acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc
      961 ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca
     1021 gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa
     1081 acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg
     1141 aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg
     1201 cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca
     1261 tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct
     1321 ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc
     1381 ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg
     1441 ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc
     1501 tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc
     1561 ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc
     1621 acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga
     1681 tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca
     1741 gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg
     1801 aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata
     1861 agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg
     1921 tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc
     1981 tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga
     2041 ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac tccgtggaaa
     2101 tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa
     2161 ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg gaagatatcg
     2221 tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg aaaacctatg
     2281 cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac accggctggg
     2341 gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc aagacaatcc
     2401 tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg atccacgacg
     2461 acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag ggcgatagcc
     2521 tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc atcctgcaga
     2581 cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg
     2641 tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac agccgcgaga
     2701 gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg aaagaacacc
     2761 ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg cagaatgggc
     2821 gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac gatgtggacc
     2881 atatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg ctgaccagaa
     2941 gcgacaagaa ccggggcaag agcgacaacg tgccctccga agaggtcgtg aagaagatga
     3001 agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag ttcgacaatc
     3061 tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc atcaagagac
     3121 agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac tcccggatga
     3181 acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc accctgaagt
     3241 ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc gagatcaaca
     3301 actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa
     3361 agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac gacgtgcgga
     3421 agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac ttcttctaca
     3481 gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag atccggaagc
     3541 ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag ggccgggatt
     3601 ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg
     3661 tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc gataagctga
     3721 tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc cccaccgtgg
     3781 cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg
     3841 tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag aatcccatcg
     3901 actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc aagctgccta
     3961 agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct gccggcgaac
     4021 tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg tacctggcca
     4081 gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag ctgtttgtgg
     4141 aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc tccaagagag
     4201 tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag caccgggata
     4261 agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc aatctgggag
     4321 cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac accagcacca
     4381 aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac gagacacgga
     4441 tcgacctgtc tcagctggga ggcgacaaaa ggccggcggc cacgaaaaag gccggccagg
     4501 caaaaaagaa aaaggaattc ggcagtggag agggcagagg aagtctgcta acatgcggtg
     4561 acgtcgagga gaatcctggc ccaatgaccg agtacaagcc cacggtgcgc ctcgccaccc
     4621 gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
     4681 cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct
     4741 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
     4801 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
     4861 cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc
     4921 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg
     4981 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
     5041 gcgccggggt gcccgccttc ctggagacct ccgcgcccca caacctcccc ttctacgagc
     5101 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca
     5161 tgacccgcaa gcccggtgcc tgagaattct aactagagct cgctgatcag cctcgactgt
     5221 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     5281 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     5341 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     5401 agagaatagc aggcatgctg gggagcggcc gcaggaaccc ctagtgatgg agttggccac
     5461 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     5521 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     5581 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     5641 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     5701 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     5761 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     5821 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     5881 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     5941 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     6001 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     6061 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     6121 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     6181 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     6241 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     6301 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     6361 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6421 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6481 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     6541 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     6601 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     6661 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     6721 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     6781 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     6841 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6901 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6961 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     7021 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     7081 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     7141 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     7201 cgtggaagcc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     7261 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     7321 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     7381 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     7441 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7501 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7561 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7621 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     7681 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7741 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7801 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7861 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7921 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7981 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     8041 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     8101 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     8161 gctcacatgt gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc
     8221 tgttagagag ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac
     8281 gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat
     8341 ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt
     8401 gtggaaagga cgaaacaccg actgggacgg aacagctttg gttttagagc tagaaatagc
     8461 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
     8521 ttgttttaga gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc
     8581 caattctgca gacaaatggc tctagaggta cccgttacat aacttacggt aaatggcccg
     8641 cctggctgac cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact
     8701 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     8761 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     8821 cattgtgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     8881 gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc
     8941 cccccctccc cacccccaat tttgtattta tttatttttt aattattttg tgc
//
