LOCUS       sequence-171355-        9015 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cas9/CRISPR plasmid for cut in human PLXNB2 gene  exon 3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 9015)
  AUTHORS   Huang Y, Tejero R, Lee VK, Brusco C, Hannah T, Bertucci TB, 
            Junqueira Alves C, Katsyv I, Kluge M, Foty R, Zhang B, Friedel CC, 
            Dai G, Zou H, Friedel RH
  TITLE     Plexin-B2 facilitates glioblastoma infiltration by modulating cell 
            biomechanics.
  JOURNAL   Commun Biol. 2021 Jan 29;4(1):145. doi: 10.1038/s42003-021-01667-4.
  PUBMED    33514835
REFERENCE   2  (bases 1 to 9015)
  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: "Commun 
            Biol."; date: "2021-01-29"; volume: "4(1):145. doi"; pages: " 
            10.1038/s42003-021-01667-4"
COMMENT     Sequence Label: pX459-sgPLXNB2
FEATURES             Location/Qualifiers
     source          1..9015
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          5..232
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      244..253
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             253..318
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             325..345
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             370..4470
                     /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             370..4470
                     /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             4471..4518
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4534..4587
                     /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             4588..5187
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4588..4607)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     complement(5215..5232)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5221..5428
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   5437..5577
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5437..5566
                     /label=AAV2 ITR
     rep_origin      5652..6107
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5739..5758)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5949..5970
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6124..6143)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6243..6265
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6303..6321)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6389..6493
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6494..6562,6563..7354)
                     /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(6712..6731)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7525..8113
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8014..8033
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        8175..8423
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     8175..8195
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     8346..8365
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        8444..8519
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        8616..8901
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
ORIGIN
        1 ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg
       61 cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc
      121 tcctccgggc tgtaattagc tgagcaagag gtaagggttt aagggatggt tggttggtgg
      181 ggtattaatg tttaattacc tggagcacct gcctgaaatc actttttttc aggttggacc
      241 ggtgccacca tggactataa ggaccacgac ggagactaca aggatcatga tattgattac
      301 aaagacgatg acgataagat ggccccaaag aagaagcgga aggtcggtat ccacggagtc
      361 ccagcagccg acaagaagta cagcatcggc ctggacatcg gcaccaactc tgtgggctgg
      421 gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc
      481 gaccggcaca gcatcaagaa gaacctgatc ggagccctgc tgttcgacag cggcgaaaca
      541 gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg
      601 atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc
      661 cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc
      721 ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg
      781 agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg
      841 gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac
      901 agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa
      961 aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag
     1021 agcagacggc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc
     1081 ggaaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg
     1141 gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg
     1201 ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac
     1261 gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc
     1321 gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc
     1381 gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc
     1441 tacgccggct acattgacgg cggagccagc caggaagagt tctacaagtt catcaagccc
     1501 atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaacag agaggacctg
     1561 ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag
     1621 ctgcacgcca ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa
     1681 aagatcgaga agatcctgac cttccgcatc ccctactacg tgggccctct ggccagggga
     1741 aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc
     1801 gaggaagtgg tggacaaggg cgcttccgcc cagagcttca tcgagcggat gaccaacttc
     1861 gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc
     1921 accgtgtata acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc
     1981 ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc tgttcaagac caaccggaaa
     2041 gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg
     2101 gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg
     2161 aaaattatca aggacaagga cttcctggac aatgaggaaa acgaggacat tctggaagat
     2221 atcgtgctga ccctgacact gtttgaggac agagagatga tcgaggaacg gctgaaaacc
     2281 tatgcccacc tgttcgacga caaagtgatg aagcagctga agcggcggag atacaccggc
     2341 tggggcaggc tgagccggaa gctgatcaac ggcatccggg acaagcagtc cggcaagaca
     2401 atcctggatt tcctgaagtc cgacggcttc gccaacagaa acttcatgca gctgatccac
     2461 gacgacagcc tgacctttaa agaggacatc cagaaagccc aggtgtccgg ccagggcgat
     2521 agcctgcacg agcacattgc caatctggcc ggcagccccg ccattaagaa gggcatcctg
     2581 cagacagtga aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac
     2641 atcgtgatcg aaatggccag agagaaccag accacccaga agggacagaa gaacagccgc
     2701 gagagaatga agcggatcga agagggcatc aaagagctgg gcagccagat cctgaaagaa
     2761 caccccgtgg aaaacaccca gctgcagaac gagaagctgt acctgtacta cctgcagaat
     2821 gggcgggata tgtacgtgga ccaggaactg gacatcaacc ggctgtccga ctacgatgtg
     2881 gaccatatcg tgcctcagag ctttctgaag gacgactcca tcgacaacaa ggtgctgacc
     2941 agaagcgaca agaaccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag
     3001 atgaagaact actggcggca gctgctgaac gccaagctga ttacccagag aaagttcgac
     3061 aatctgacca aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag
     3121 agacagctgg tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg
     3181 atgaacacta agtacgacga gaatgacaag ctgatccggg aagtgaaagt gatcaccctg
     3241 aagtccaagc tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc
     3301 aacaactacc accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc
     3361 aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg
     3421 cggaagatga tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc
     3481 tacagcaaca tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg
     3541 aagcggcctc tgatcgagac aaacggcgaa accggggaga tcgtgtggga taagggccgg
     3601 gattttgcca ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt gaaaaagacc
     3661 gaggtgcaga caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgataag
     3721 ctgatcgcca gaaagaagga ctgggaccct aagaagtacg gcggcttcga cagccccacc
     3781 gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag
     3841 agtgtgaaag agctgctggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc
     3901 atcgactttc tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg
     3961 cctaagtact ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccggc
     4021 gaactgcaga agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg
     4081 gccagccact atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt
     4141 gtggaacagc acaagcacta cctggacgag atcatcgagc agatcagcga gttctccaag
     4201 agagtgatcc tggccgacgc taatctggac aaagtgctgt ccgcctacaa caagcaccgg
     4261 gataagccca tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg
     4321 ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc
     4381 accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca
     4441 cggatcgacc tgtctcagct gggaggcgac aaaaggccgg cggccacgaa aaaggccggc
     4501 caggcaaaaa agaaaaagga attcggcagt ggagagggca gaggaagtct gctaacatgc
     4561 ggtgacgtcg aggagaatcc tggcccaatg accgagtaca agcccacggt gcgcctcgcc
     4621 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     4681 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     4741 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     4801 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     4861 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     4921 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     4981 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     5041 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccacaacct ccccttctac
     5101 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     5161 tgcatgaccc gcaagcccgg tgcctgagaa ttctaactag agctcgctga tcagcctcga
     5221 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     5281 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     5341 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     5401 gggaagagaa tagcaggcat gctggggagc ggccgcagga acccctagtg atggagttgg
     5461 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     5521 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     5581 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     5641 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     5701 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     5761 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     5821 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     5881 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     5941 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     6001 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     6061 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     6121 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     6181 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     6241 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     6301 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     6361 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     6421 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     6481 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     6541 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     6601 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     6661 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     6721 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     6781 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     6841 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     6901 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     6961 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     7021 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     7081 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     7141 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     7201 tgagcgtgga agccgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     7261 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     7321 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     7381 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     7441 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     7501 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     7561 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     7621 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     7681 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     7741 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     7801 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     7861 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     7921 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     7981 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     8041 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     8101 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     8161 ttttgctcac atgtgagggc ctatttccca tgattccttc atatttgcat atacgataca
     8221 aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata ttagtacaaa
     8281 atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta
     8341 aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata
     8401 tcttgtggaa aggacgaaac accgttctcg gcggcgaccg tcagttttag agctagaaat
     8461 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
     8521 tttttgtttt agagctagaa atagcaagtt aaaataaggc tagtccgttt ttagcgcgtg
     8581 cgccaattct gcagacaaat ggctctagag gtacccgtta cataacttac ggtaaatggc
     8641 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caatagtaac gccaataggg
     8701 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     8761 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     8821 tggcattgtg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     8881 ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc
     8941 tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg
     9001 atgggggcgg ggggg
//
