LOCUS       sequence-204106-        9120 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  sgRNA targeting murine CD90.1. Co-expresses SpCas9-2A-EGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9120)
  AUTHORS   Kornete M, Marone R, Jeker LT
  TITLE     Highly Efficient and Versatile Plasmid-Based Gene Editing in Primary
            T Cells.
  JOURNAL   J Immunol. 2018 Feb 14. pii: jimmunol.1701121. doi: 
            10.4049/jimmunol.1701121.
  PUBMED    29445007
REFERENCE   2  (bases 1 to 9120)
  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: "J Immunol. 
            2018 Feb 14. pii: jimmunol.1701121. doi: 10.4049/jimmunol.1701121."
COMMENT     Sequence Label: p163_LTJ_sgRNACD90.1
FEATURES             Location/Qualifiers
     source          1..9120
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      231..240
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             240..305
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             312..332
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             357..4457
                     /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             357..4457
                     /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             4458..4505
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4521..4574
                     /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(4575..4577,4578..5288)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4575..4577,4578..5288)
                     /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(4878..4897)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5225..5246
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5316..5333)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5322..5529
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5538..5678
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5538..5667
                     /label=AAV2 ITR
     rep_origin      5753..6208
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5840..5859)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6050..6071
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6225..6244)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6344..6366
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6404..6422)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6490..6594
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6595..6663,6664..7455)
                     /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(6813..6832)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7626..8214
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8115..8134
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8276..8524
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8276..8296
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8447..8466
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8545..8620
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8717..9002
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc
       61 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
      121 aattagctga gcaagaggta agggtttaag ggatggttgg ttggtggggt attaatgttt
      181 aattacctgg agcacctgcc tgaaatcact ttttttcagg ttggaccggt gccaccatgg
      241 actataagga ccacgacgga gactacaagg atcatgatat tgattacaaa gacgatgacg
      301 ataagatggc cccaaagaag aagcggaagg tcggtatcca cggagtccca gcagccgaca
      361 agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg
      421 acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca
      481 tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc
      541 ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc
      601 aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag
      661 agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg
      721 tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg
      781 tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca
      841 agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca
      901 agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg
      961 ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg
     1021 aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg
     1081 ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca
     1141 aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg
     1201 gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga
     1261 gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca
     1321 agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc
     1381 tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca
     1441 ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga
     1501 tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc
     1561 ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg cacgccattc
     1621 tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga
     1681 tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg
     1741 cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg
     1801 acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc
     1861 ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg
     1921 agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg
     1981 agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc
     2041 agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg
     2101 tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg
     2161 acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc
     2221 tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt
     2281 tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga
     2341 gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc
     2401 tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga
     2461 cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc
     2521 acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg
     2581 tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa
     2641 tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc
     2701 ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa
     2761 acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt
     2821 acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc
     2881 ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga
     2941 accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact
     3001 ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg
     3061 ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg
     3121 aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt
     3181 acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg
     3241 tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc
     3301 acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta
     3361 agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg
     3421 ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca
     3481 tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga
     3541 tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg
     3601 tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag
     3661 gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa
     3721 agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg
     3781 tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc
     3841 tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg
     3901 aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc
     3961 tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg
     4021 gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg
     4081 agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca
     4141 agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg
     4201 ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca
     4261 gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg
     4321 ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc
     4381 tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt
     4441 ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga
     4501 aaaaggaatt cggcagtgga gagggcagag gaagtctgct aacatgcggt gacgtcgagg
     4561 agaatcctgg cccagtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     4621 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     4681 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     4741 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     4801 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     4861 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     4921 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     4981 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     5041 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     5101 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     5161 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     5221 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     5281 tgtacaagga attctaacta gagctcgctg atcagcctcg actgtgcctt ctagttgcca
     5341 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     5401 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     5461 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaga atagcaggca
     5521 tgctggggag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     5581 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     5641 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     5701 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     5761 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     5821 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     5881 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     5941 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     6001 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     6061 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     6121 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     6181 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     6241 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     6301 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     6361 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     6421 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     6481 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     6541 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     6601 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     6661 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     6721 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     6781 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     6841 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     6901 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6961 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     7021 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     7081 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     7141 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     7201 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     7261 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg aagccgcggt
     7321 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     7381 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     7441 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     7501 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     7561 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     7621 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     7681 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     7741 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     7801 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     7861 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     7921 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7981 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     8041 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     8101 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     8161 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     8221 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgtgaggg
     8281 cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta gagagataat
     8341 tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta
     8401 ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct
     8461 taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa
     8521 caccgtttgt gagcttcgag tctcgtttta gagctagaaa tagcaagtta aaataaggct
     8581 agtccgttat caacttgaaa aagtggcacc gagtcggtgc ttttttgttt tagagctaga
     8641 aatagcaagt taaaataagg ctagtccgtt tttagcgcgt gcgccaattc tgcagacaaa
     8701 tggctctaga ggtacccgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     8761 acgacccccg cccattgacg tcaatagtaa cgccaatagg gactttccat tgacgtcaat
     8821 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     8881 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattgt gcccagtaca
     8941 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     9001 tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc
     9061 caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg
//
