LOCUS       sequence-218029-        9256 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  sgRNA targeting murine Foxp3 scurfy. Co-expresses SpCas9-2A-EGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9256)
  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 9256)
  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: p236_LTJ_sgRNAFoxp3sf/J
FEATURES             Location/Qualifiers
     source          1..9256
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          128..355
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      367..376
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             376..441
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             448..468
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             493..4593
                     /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             493..4593
                     /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             4594..4641
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4657..4710
                     /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(4711..4713,4714..5424)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(4711..4713,4714..5424)
                     /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(5014..5033)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5361..5382
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5452..5469)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5458..5665
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5674..5814
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5674..5803
                     /label=AAV2 ITR
     rep_origin      5889..6344
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5976..5995)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6186..6207
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6361..6380)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6480..6502
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6540..6558)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6626..6730
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6731..6799,6800..7591)
                     /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(6949..6968)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7762..8350
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8251..8270
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8412..8660
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8412..8432
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8583..8602
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8681..8756
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8853..9138
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg
       61 aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg
      121 gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg
      181 ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc
      241 ttctcctccg ggctgtaatt agctgagcaa gaggtaaggg tttaagggat ggttggttgg
      301 tggggtatta atgtttaatt acctggagca cctgcctgaa atcacttttt ttcaggttgg
      361 accggtgcca ccatggacta taaggaccac gacggagact acaaggatca tgatattgat
      421 tacaaagacg atgacgataa gatggcccca aagaagaagc ggaaggtcgg tatccacgga
      481 gtcccagcag ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc
      541 tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac
      601 accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa
      661 acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac
      721 cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc
      781 ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc
      841 atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac
      901 ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc
      961 ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac
     1021 aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag
     1081 gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc
     1141 aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg
     1201 ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac
     1261 ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac
     1321 ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc
     1381 gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg
     1441 agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct
     1501 ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac
     1561 ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag
     1621 cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac
     1681 ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga
     1741 gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg
     1801 gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg
     1861 ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac
     1921 ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac
     1981 ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac
     2041 ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc
     2101 gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg
     2161 aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc
     2221 gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg
     2281 ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa
     2341 gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa
     2401 acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc
     2461 ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag
     2521 acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc
     2581 cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc
     2641 gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc
     2701 ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag
     2761 aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc
     2821 cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa
     2881 gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag
     2941 aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat
     3001 gtggaccata tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg
     3061 accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag
     3121 aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc
     3181 gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc
     3241 aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc
     3301 cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc
     3361 ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag
     3421 atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg
     3481 atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac
     3541 gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc
     3601 ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc
     3661 cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc
     3721 cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag
     3781 accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat
     3841 aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc
     3901 accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg
     3961 aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat
     4021 cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag
     4081 ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc
     4141 ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac
     4201 ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg
     4261 tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc
     4321 aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac
     4381 cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat
     4441 ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc
     4501 agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag
     4561 acacggatcg acctgtctca gctgggaggc gacaaaaggc cggcggccac gaaaaaggcc
     4621 ggccaggcaa aaaagaaaaa ggaattcggc agtggagagg gcagaggaag tctgctaaca
     4681 tgcggtgacg tcgaggagaa tcctggccca gtgagcaagg gcgaggagct gttcaccggg
     4741 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     4801 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     4861 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     4921 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     4981 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     5041 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     5101 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     5161 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     5221 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     5281 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     5341 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     5401 ctcggcatgg acgagctgta caaggaattc taactagagc tcgctgatca gcctcgactg
     5461 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     5521 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     5581 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     5641 aagagaatag caggcatgct ggggagcggc cgcaggaacc cctagtgatg gagttggcca
     5701 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     5761 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     5821 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     5881 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     5941 cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct tcccttcctt
     6001 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     6061 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     6121 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     6181 taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg gctattcttt
     6241 tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc tgatttaaca
     6301 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     6361 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     6421 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     6481 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     6541 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     6601 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     6661 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6721 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     6781 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     6841 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     6901 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     6961 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     7021 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     7081 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     7141 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     7201 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     7261 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     7321 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     7381 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     7441 gcgtggaagc cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     7501 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     7561 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     7621 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     7681 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     7741 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     7801 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     7861 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     7921 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7981 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     8041 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     8101 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     8161 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     8221 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     8281 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     8341 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     8401 tgctcacatg tgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
     8461 ctgttagaga gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata
     8521 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
     8581 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
     8641 tgtggaaagg acgaaacacc gagagctctt ttgtccattg gttttagagc tagaaatagc
     8701 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
     8761 ttgttttaga gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc
     8821 caattctgca gacaaatggc tctagaggta cccgttacat aacttacggt aaatggcccg
     8881 cctggctgac cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact
     8941 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     9001 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     9061 cattgtgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     9121 gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc
     9181 cccccctccc cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg
     9241 ggggcggggg gggggg
//
