LOCUS       sequence-204109-        9129 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  sgRNA targeting murine Foxp3 mutant. Co-expresses SpCas9-2A-EGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9129)
  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 9129)
  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: p240_LTJ_sgRNAFoxp3K276X
FEATURES             Location/Qualifiers
     source          1..9129
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      239..248
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             248..313
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             320..340
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             365..4465
                     /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             365..4465
                     /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             4466..4513
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4529..4582
                     /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(4583..4585,4586..5296)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4583..4585,4586..5296)
                     /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(4886..4905)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5233..5254
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5324..5341)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5330..5537
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5546..5686
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5546..5675
                     /label=AAV2 ITR
     rep_origin      5761..6216
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5848..5867)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6058..6079
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6233..6252)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6352..6374
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6412..6430)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6498..6602
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6603..6671,6672..7463)
                     /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(6821..6840)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7634..8222
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8123..8142
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8284..8532
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8284..8304
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8455..8474
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8553..8628
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8725..9010
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
       61 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      121 cgggctgtaa ttagctgagc aagaggtaag ggtttaaggg atggttggtt ggtggggtat
      181 taatgtttaa ttacctggag cacctgcctg aaatcacttt ttttcaggtt ggaccggtgc
      241 caccatggac tataaggacc acgacggaga ctacaaggat catgatattg attacaaaga
      301 cgatgacgat aagatggccc caaagaagaa gcggaaggtc ggtatccacg gagtcccagc
      361 agccgacaag aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt
      421 gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg
      481 gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga
      541 ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg
      601 ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag
      661 actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg
      721 caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa
      781 gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca
      841 catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga
      901 cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc
      961 catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag
     1021 acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa
     1081 cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga
     1141 ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc
     1201 ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat
     1261 cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc
     1321 tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg
     1381 gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc
     1441 cggctacatt gacggcggag ccagccagga agagttctac aagttcatca agcccatcct
     1501 ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg
     1561 gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca
     1621 cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat
     1681 cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag
     1741 cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga
     1801 agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa
     1861 gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt
     1921 gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct
     1981 gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac
     2041 cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat
     2101 ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat
     2161 tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt
     2221 gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc
     2281 ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg
     2341 caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct
     2401 ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga
     2461 cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct
     2521 gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac
     2581 agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt
     2641 gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag
     2701 aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc
     2761 cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg
     2821 ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca
     2881 tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag
     2941 cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa
     3001 gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct
     3061 gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca
     3121 gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa
     3181 cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc
     3241 caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa
     3301 ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa
     3361 gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa
     3421 gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag
     3481 caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg
     3541 gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt
     3601 tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt
     3661 gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat
     3721 cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc
     3781 ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt
     3841 gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga
     3901 ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa
     3961 gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact
     4021 gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag
     4081 ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga
     4141 acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt
     4201 gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa
     4261 gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc
     4321 ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa
     4381 agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat
     4441 cgacctgtct cagctgggag gcgacaaaag gccggcggcc acgaaaaagg ccggccaggc
     4501 aaaaaagaaa aaggaattcg gcagtggaga gggcagagga agtctgctaa catgcggtga
     4561 cgtcgaggag aatcctggcc cagtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     4621 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     4681 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     4741 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     4801 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     4861 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     4921 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     4981 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     5041 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     5101 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     5161 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     5221 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     5281 ggacgagctg tacaaggaat tctaactaga gctcgctgat cagcctcgac tgtgccttct
     5341 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     5401 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     5461 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagagaat
     5521 agcaggcatg ctggggagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     5581 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     5641 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     5701 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     5761 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     5821 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     5881 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     5941 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     6001 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     6061 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     6121 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     6181 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     6241 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     6301 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     6361 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     6421 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     6481 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     6541 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     6601 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     6661 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     6721 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6781 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     6841 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     6901 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     6961 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     7021 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     7081 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     7141 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     7201 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     7261 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggaa
     7321 gccgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     7381 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     7441 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     7501 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     7561 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     7621 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     7681 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     7741 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     7801 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     7861 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     7921 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     7981 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     8041 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     8101 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     8161 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     8221 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     8281 tgtgagggcc tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga
     8341 gagataattg gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt
     8401 agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat
     8461 catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa
     8521 ggacgaaaca ccgcaagata tctagtccat tggttttaga gctagaaata gcaagttaaa
     8581 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt ttttgtttta
     8641 gagctagaaa tagcaagtta aaataaggct agtccgtttt tagcgcgtgc gccaattctg
     8701 cagacaaatg gctctagagg tacccgttac ataacttacg gtaaatggcc cgcctggctg
     8761 accgcccaac gacccccgcc cattgacgtc aatagtaacg ccaataggga ctttccattg
     8821 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     8881 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattgtgc
     8941 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     9001 tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc
     9061 cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg
     9121 ggggggggg
//
