LOCUS       sequence-240490-        9113 bp    DNA     linear   UNA 15-APR-2025
DEFINITION  sgRNA against human Thy-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9113)
  TITLE     Vizler lab CRISPR plasmids
REFERENCE   2  (bases 1 to 9113)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: Thy-1-CRISPR-gRNA#2-(pSpCas9(BB)-2A-Puro (PX45...
FEATURES             Location/Qualifiers
     source          1..9113
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          102..329
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      341..350
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             350..415
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             422..442
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             467..4567
                     /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             467..4567
                     /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             4568..4615
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4631..4684
                     /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             4685..5284
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4685..4704)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5181..5201
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5312..5329)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5318..5525
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5534..5674
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5534..5663
                     /label=AAV2 ITR
     rep_origin      5749..6204
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5836..5855)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6046..6067
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6221..6240)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6340..6362
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6400..6418)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6486..6590
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6591..6659,6660..7451)
                     /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(6809..6828)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7622..8210
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8111..8130
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8272..8520
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8272..8292
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8443..8462
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8542..8617
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8714..8999
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc
       61 ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc
      121 ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg
      181 cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagctga
      241 gcaagaggta agggtttaag ggatggttgg ttggtggggt attaatgttt aattacctgg
      301 agcacctgcc tgaaatcact ttttttcagg ttggaccggt gccaccatgg actataagga
      361 ccacgacgga gactacaagg atcatgatat tgattacaaa gacgatgacg ataagatggc
      421 cccaaagaag aagcggaagg tcggtatcca cggagtccca gcagccgaca agaagtacag
      481 catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa
      541 ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa
      601 cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag
      661 aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt
      721 cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag agtccttcct
      781 ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt
      841 ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac
      901 cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg
      961 ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat
     1021 ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt
     1081 ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat
     1141 cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct
     1201 gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct
     1261 gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta
     1321 cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct
     1381 gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga
     1441 cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa
     1501 gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg
     1561 agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac
     1621 cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga
     1681 caacggcagc atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca
     1741 ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt
     1801 ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac
     1861 cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc
     1921 ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa
     1981 ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa
     2041 agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa
     2101 ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga
     2161 ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg
     2221 gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt
     2281 cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt
     2341 tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa
     2401 agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct
     2461 gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga
     2521 cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga
     2581 ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa
     2641 tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga
     2701 gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga
     2761 gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga
     2821 gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct
     2881 gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca
     2941 ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt
     3001 tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa
     3061 gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct
     3121 gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg
     3181 cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca
     3241 gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa
     3301 tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt
     3361 ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga
     3421 cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag
     3481 cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga
     3541 gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt
     3601 caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa
     3661 cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt
     3721 gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag
     3781 caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg
     3841 ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt
     3901 ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat
     3961 caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg
     4021 ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct
     4081 ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact
     4141 ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa
     4201 gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct
     4261 ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa
     4321 tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc
     4381 cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta
     4441 ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac
     4501 cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg
     4561 aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga aaaaggaatt
     4621 cggcagtgga gagggcagag gaagtctgct aacatgcggt gacgtcgagg agaatcctgg
     4681 cccaatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
     4741 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
     4801 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     4861 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     4921 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     4981 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     5041 ggagcccgcg tggttcctgg ccaccgtcgg agtctcgccc gaccaccagg gcaagggtct
     5101 gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt
     5161 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     5221 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     5281 ctgagaattc taactagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     5341 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     5401 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     5461 gggggtgggg tggggcagga cagcaagggg gaggattggg aagagaatag caggcatgct
     5521 ggggagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     5581 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     5641 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     5701 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     5761 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5821 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5881 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     5941 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     6001 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     6061 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     6121 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     6181 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     6241 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     6301 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     6361 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     6421 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     6481 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6541 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     6601 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     6661 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     6721 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     6781 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     6841 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     6901 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     6961 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     7021 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     7081 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     7141 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     7201 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     7261 cggctggctg gtttattgct gataaatctg gagccggtga gcgtggaagc cgcggtatca
     7321 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     7381 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     7441 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     7501 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     7561 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     7621 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7681 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7741 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     7801 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7861 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7921 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7981 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     8041 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     8101 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     8161 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     8221 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg tgagggccta
     8281 tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga gataattgga
     8341 attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa
     8401 tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc
     8461 gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc
     8521 gaggctgaac tcgtactgga tgttttagag ctagaaatag caagttaaaa taaggctagt
     8581 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttgttttag agctagaaat
     8641 agcaagttaa aataaggcta gtccgttttt agcgcgtgcg ccaattctgc agacaaatgg
     8701 ctctagaggt acccgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     8761 acccccgccc attgacgtca atagtaacgc caatagggac tttccattga cgtcaatggg
     8821 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     8881 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattgtgcc cagtacatga
     8941 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     9001 tcgaggtgag ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa
     9061 ttttgtattt atttattttt taattatttt gtgcagcgat gggggcgggg ggg
//
