LOCUS       sequence-489390-       14676 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Variant of lentiCRISPRv2 that confers blasticidin S resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14676)
  AUTHORS   Stringer BW, Day BW, D'Souza RCJ, Jamieson PR, Ensbey KS, Bruce ZC, 
            Lim YC, Goasdoue K, Offenhauser C, Akgul S, Allan S, Robertson T, 
            Lucas P, Tollesson G, Campbell S, Winter C, Do H, Dobrovic A, Inglis
            PL, Jeffree RL, Johns TG, Boyd AW
  TITLE     A reference collection of patient-derived cell line and xenograft 
            models of proneural, classical and mesenchymal glioblastoma.
  JOURNAL   Sci Rep. 2019 Mar 20;9(1):4902. doi: 10.1038/s41598-019-41277-z.
  PUBMED    30894629
REFERENCE   2  (bases 1 to 14676)
  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: "Sci Rep."; 
            date: "2019-03-20"; volume: "9(1):4902. doi"; pages: " 
            10.1038/s41598-019-41277-z"
COMMENT     Sequence Label: lentiCRISPRv2 blast
FEATURES             Location/Qualifiers
     source          1..14676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(277..525)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(335..354)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(505..525)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    576..693
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(991..1035)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1220..1453
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    1946..2071
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             2118..2298
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(2316..2514)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2341..2361)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2516..2895
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3071..3090
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3165..3269
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3270..3338,3339..4130)
                     /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(3488..3507)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4301..4889
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4790..4809
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5043..5060
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5177..5198
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5213..5218,5219..5236,5237..5243)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5251..5267
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5256..5278
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5275..5291
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5275..5291
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5340..5461
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5351..5370)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5405..5424
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5591..5965)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(5984..6031)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(6079..6408)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6093..6228
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6147..6166)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6393..6413
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(6422..6850)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6532..6553)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6744..6763
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    6896..7120
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     7109..7126
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     LTR             7152..7385
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    7457..8045
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             7563..7574
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     7972..7992
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8061..8456)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="AKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(8457..8513)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(8523..8546)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(8547..8594)
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             complement(8595..12698)
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(8595..12698)
                     /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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     regulatory      12695..12704
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(12723..12934)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_RNA        complement(12996..13071)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 tttgaagatg ctcaaaaaat gattgaagca ggagctgacc gcttaggaac aagtggtgga
       61 gtagctatta ttaaaggtga agaaaacaac gcgagttact aaaactagcg tttttttatt
      121 ttgctcattt ttattaaaag tttgcaaaaa ggaacataaa aattctaatt attgatacta
      181 aagttattaa aaagaagatt ttggttgatt ttataaaggt catagaatat aatattttag
      241 catgtgtatt ttgtgtgctc atttacaacc gtctccggtg tttcgtcctt tccacaagat
      301 atataaagcc aagaaatcga aatactttca agttacggta agcatatgat agtccatttt
      361 aaaacataat tttaaaactg caaactaccc aagaaattat tactttctac gtcacgtatt
      421 ttgtactaat atctttgtgt ttacagtcaa attaattcta attatctctc taacagcctt
      481 gtatcgtata tgcaaatatg aaggaatcat gggaaatagg ccctcggtac cttaattaac
      541 caaactggat ctctgctgtc cctgtaataa acccgaaaat tttgaatttt tgtaatttgt
      601 ttttgtaatt ctttagtttg tatgtctgtt gctattatgt ctactattct ttcccctgca
      661 ctgtaccccc caatcccccc ttttctttta aaattgtgga tgaatactgc catttgtctg
      721 cagaattggc gcacgcagtg ccgatccgtt cactaatcga atggatctgt ctctgtctct
      781 ctctccacct tcttcttcta ttccttcggg cctgtcgggt cccctcgggg ttgggaggtg
      841 ggtctgaaac gataatggtg aatatccctg cctaactcta ttcactatag aaagtacagc
      901 aaaaactatt cttaaaccta ccaagcctcc tactatcatt atgaataatt ttatatacca
      961 cagccaattt gttatgttaa accaattcca caaacttgcc catttatcta attccaataa
     1021 ttcttgttca ttcttttctt gctggttttg cgattcttca attaaggagt gtattaagct
     1081 tgtgtaattg ttaatttctc tgtcccactc catccaggtc gtgtgattcc aaatctgttc
     1141 cagagattta ttactccaac tagcattcca aggcacagca gtggtgcaaa tgagttttcc
     1201 agagcaaccc caaatcccca ggagctgttg atcctttagg tatctttcca cagccaggat
     1261 tcttgcctgg agctgcttga tgccccagac tgtgagttgc aacagatgct gttgcgcctc
     1321 aatagccctc agcaaattgt tctgctgctg cactatacca gacaataatt gtctggcctg
     1381 taccgtcagc gtcattgacg ctgcgcccat agtgcttcct gctgctccca agaacccaag
     1441 gaacaaagct cctattccca ctgctctttt ttctctctgc accactcttc tctttgcctt
     1501 ggtgggtgct actcctaatg gttcaatttt tactacttta tatttatata attcacttct
     1561 ccaattgtcc ctcatatctc ctcctccagg tctgaagatc agcggccgct tgctgtgcgg
     1621 tggtcttact tttgttttgc tcttcctcta tcttgtctaa agcttccttg gtgtctttta
     1681 tctctatcct ttgatgcaca caatagaggg ttgctactgt attatataat gatctaagtt
     1741 cttctgatcc tgtctgaagg gatggttgta gctgtcccag tatttgtcta cagccttctg
     1801 atgtttctaa caggccagga ttaactgcga atcgttctag ctccctgctt gcccatacta
     1861 tatgttttaa tttatatttt ttctttcccc ctggccttaa ccgaattttt tcccatcgcg
     1921 atctaattct cccccgctta atactgacgc tctcgcaccc atctctctcc ttctagcctc
     1981 cgctagtcaa aatttttggc gtactcacca gtcgccgccc ctcgcctctt gccgtgcgcg
     2041 cttcagcaag ccgagtcctg cgtcgagaga gctcctctgg tttccctttc gctttcaagt
     2101 ccctgttcgg gcgccactgc tagagatttt ccacactgac taaaagggtc tgagggatct
     2161 ctagttacca gagtcacaca acagacgggc acacactact tgaagcactc aaggcaagct
     2221 ttattgaggc ttaagcagtg ggttccctag ttagccagag agctcccagg ctcagatctg
     2281 gtctaaccag agagacccag tacaggcaaa acgcgctgct tatatagacc tcccaccgta
     2341 cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc
     2401 gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc
     2461 cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg
     2521 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt
     2581 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg
     2641 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
     2701 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
     2761 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
     2821 ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat taataactag
     2881 tcaataatca atgtcaacgc gcgcgtatat ctggcccgta catcgcgaag cagcgcaaaa
     2941 cgcctaaccc taagcagatt cttcatgcaa ttgtcggtca agccttgcct tgttgtagct
     3001 taaattttgc tcgcgcacta ctcagcgacc tccaacacac aagcagggag cagatactgg
     3061 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatagggg atcgggagat
     3121 ctcccgatcc gtcgacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3181 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3241 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     3301 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3361 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     3421 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     3481 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     3541 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     3601 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     3661 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     3721 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     3781 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     3841 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     3901 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     3961 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4021 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4081 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     4141 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4201 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4261 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4321 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4381 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4441 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4501 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4561 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4621 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4681 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4741 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4801 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4861 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4921 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     4981 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     5041 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     5101 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     5161 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     5221 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     5281 acagctatga ccatgattac gccaagctct agctagaggt cgacggtata cagacatgat
     5341 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     5401 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     5461 tggggtgggc gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt
     5521 cccggaaaac gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc
     5581 gtagcacgtg tcagtcctgc tcctcggcca cgaagtgcac gcagttgccg gccgggtcgc
     5641 gcagggcgaa ctcccgcccc cacggctgct cgccgatctc ggtcatggcc ggcccggagg
     5701 cgtcccggaa gttcgtggac acgacctccg accactcggc gtacagctcg tccaggccgc
     5761 gcacccacac ccaggccagg gtgttgtccg gcaccacctg gtcctggacc gcgctgatga
     5821 acagggtcac gtcgtcccgg accacaccgg cgaagtcgtc ctccacgaag tcccgggaga
     5881 acccgagccg gtcggtccag aactcgaccg ctccggcgac gtcgcgcgcg gtgagcaccg
     5941 gaacggcact ggtcaacttg gccatggttt agttcctcac cttgtcgtat tatactatgc
     6001 cgatatacta tgccgatgat taattgtcaa cacgtgctga tcagatccga aaatggatat
     6061 acaagctccc gggagctttt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt
     6121 ctggaatagc tcagaggcag aggcggcctc ggcctctgca taaataaaaa aaattagtca
     6181 gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg gcggagttag
     6241 gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
     6301 ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc tttgcatact
     6361 tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat tccacagaat
     6421 taattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     6481 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     6541 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     6601 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     6661 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     6721 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     6781 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     6841 cagggcgcgt ggggataccc cctagagccc cagctggttc tttccgcctc agaagccata
     6901 gagcccaccg catccccagc atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc
     6961 tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga tgcaatttcc
     7021 tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga aggcacgggg
     7081 gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcgaggctga tcagcgggtt
     7141 taaacgggcc ctgctagaga ttttccacac tgactaaaag ggtctgaggg atctctagtt
     7201 accagagtca cacaacagac gggcacacac tacttgaagc actcaaggca agctttattg
     7261 aggcttaagc agtgggttcc ctagttagcc agagagctcc caggctcaga tctggtctaa
     7321 ccagagagac ccagtacaag caaaaagcag atcttgtctt cgttgggagt gaattagccc
     7381 ttccagtccc cccttttctt ttaaaaagtg gctaagatct acagctgcct tgtaagtcat
     7441 tggtcttaaa gtcgacgcgg ggaggcggcc caaagggaga tccgactcgt ctgagggcga
     7501 aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc cgcgggaagg aaggtccgct
     7561 ggattgaggg ccgaagggac gtagcagaag gacgtcccgc gcagaatcca ggtggcaaca
     7621 caggcgagca gccaaggaaa ggacgatgat ttccccgaca acaccacgga attgtcagtg
     7681 cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag ttccgccgtg
     7741 gcaataggga gggggaaagc gaaagtcccg gaaaggagct gacaggtggt ggcaatgccc
     7801 caaccagtgg gggttgcgtc agcaaacaca gtgcacacca cgccacgttg cctgacaacg
     7861 ggccacaact cctcataaag agacagcaac caggatttat acaaggagga gaaaatgaaa
     7921 gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc acatagcgta
     7981 aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta atccagaggt
     8041 tgattgtcga cttaacgcgt ttagccctcc cacacataac cagagggcag caattcacga
     8101 atcccaactg ccgtcggctg tccatcactg tccttcacta tggctttgat cccaggatgc
     8161 agatcgagaa gcacctgtcg gcaccgtccg caggggctca agatgcccct gttctcattt
     8221 ccgatcgcga cgatacaagt caggttgcca gctgccgcag cagcagcagt gcccagcacc
     8281 acgagttctg cacaaggtcc cccagtaaaa tgatatacat tgacaccagt gaagatgcgg
     8341 ccgtcgctag agagagctgc gctggcgacg ctgtagtctt cagagatggg gatgctgttg
     8401 attgtagccg ttgctctttc aatgagggtg gattcttctt gagacaaagg cttggccggt
     8461 ccaggattct cttcgacatc tccggcttgt ttcagcagag agaagtttgt tgcgccggat
     8521 cccttatcgt catcgtcttt gtaatctttc ttcttcttag cctgtccagc cttctttgtg
     8581 gcggcaggtc gcttgtcgcc tcccagctga gacaggtcga tccgtgtctc gtacaggccg
     8641 gtgatgctct ggtggatcag ggtggcgtcc agcacctctt tggtgctggt gtacctcttc
     8701 cggtcgatgg tggtgtcaaa gtacttgaag gcggcagggg ctcccagatt ggtcagggta
     8761 aacaggtgga tgatattctc ggcctgctct ctgatgggct tatcccggtg cttgttgtag
     8821 gcggacagca ctttgtccag attagcgtcg gccaggatca ctctcttgga gaactcgctg
     8881 atctgctcga tgatctcgtc caggtagtgc ttgtgctgtt ccacaaacag ctgtttctgc
     8941 tcattatcct cgggggagcc cttcagcttc tcatagtggc tggccaggta caggaagttc
     9001 acatatttgg agggcagggc cagttcgttt cccttctgca gttcgccggc agaggccagc
     9061 attctcttcc ggccgttttc cagctcgaac agggagtact taggcagctt gatgatcagg
     9121 tcctttttca cttctttgta gcccttggct tccagaaagt cgatgggatt cttctcgaag
     9181 ctgcttcttt ccatgatggt gatccccagc agctctttca cactcttcag tttcttggac
     9241 ttgccctttt ccactttggc caccaccagc acagaatagg ccacggtggg gctgtcgaag
     9301 ccgccgtact tcttagggtc ccagtccttc tttctggcga tcagcttatc gctgttcctc
     9361 ttgggcagga tagactcttt gctgaagccg cctgtctgca cctcggtctt tttcacgata
     9421 ttcacttggg gcatgctcag cactttccgc acggtggcaa aatcccggcc cttatcccac
     9481 acgatctccc cggtttcgcc gtttgtctcg atcagaggcc gcttccggat ctcgccgttg
     9541 gccagggtaa tctcggtctt gaaaaagttc atgatgttgc tgtagaagaa gtacttggcg
     9601 gtagccttgc cgatttcctg ctcgctcttg gcgatcatct tccgcacgtc gtacaccttg
     9661 tagtcgccgt acacgaactc gctttccagc ttagggtact ttttgatcag ggcggttccc
     9721 acgacggcgt tcaggtaggc gtcgtgggcg tggtggtagt tgttgatctc gcgcactttg
     9781 taaaactgga aatccttccg gaaatcggac accagcttgg acttcagggt gatcactttc
     9841 acttcccgga tcagcttgtc attctcgtcg tacttagtgt tcatccggga gtccaggatc
     9901 tgtgccacgt gctttgtgat ctgccgggtt tccaccagct gtctcttgat gaagccggcc
     9961 ttatccagtt cgctcaggcc gcctctctcg gccttggtca gattgtcgaa ctttctctgg
    10021 gtaatcagct tggcgttcag cagctgccgc cagtagttct tcatcttctt cacgacctct
    10081 tcggagggca cgttgtcgct cttgccccgg ttcttgtcgc ttctggtcag caccttgttg
    10141 tcgatggagt cgtccttcag aaagctctga ggcacgatat ggtccacatc gtagtcggac
    10201 agccggttga tgtccagttc ctggtccacg tacatatccc gcccattctg caggtagtac
    10261 aggtacagct tctcgttctg cagctgggtg ttttccacgg ggtgttcttt caggatctgg
    10321 ctgcccagct ctttgatgcc ctcttcgatc cgcttcattc tctcgcggct gttcttctgt
    10381 cccttctggg tggtctggtt ctctctggcc atttcgatca cgatgttctc gggcttgtgc
    10441 cggcccatca ctttcacgag ctcgtccacc accttcactg tctgcaggat gcccttctta
    10501 atggcggggc tgccggccag attggcaatg tgctcgtgca ggctatcgcc ctggccggac
    10561 acctgggctt tctggatgtc ctctttaaag gtcaggctgt cgtcgtggat cagctgcatg
    10621 aagtttctgt tggcgaagcc gtcggacttc aggaaatcca ggattgtctt gccggactgc
    10681 ttgtcccgga tgccgttgat cagcttccgg ctcagcctgc cccagccggt gtatctccgc
    10741 cgcttcagct gcttcatcac tttgtcgtcg aacaggtggg cataggtttt cagccgttcc
    10801 tcgatcatct ctctgtcctc aaacagtgtc agggtcagca cgatatcttc cagaatgtcc
    10861 tcgttttcct cattgtccag gaagtccttg tccttgataa ttttcagcag atcgtggtat
    10921 gtgcccaggg aggcgttgaa ccgatcttcc acgccggaga tttccacgga gtcgaagcac
    10981 tcgattttct tgaagtagtc ctctttcagc tgcttcacgg tcactttccg gttggtcttg
    11041 aacagcaggt ccacgatggc ctttttctgc tcgccgctca ggaaggcggg ctttctcatt
    11101 ccctcggtca cgtatttcac tttggtcagc tcgttataca cggtgaagta ctcgtacagc
    11161 aggctgtgct tgggcagcac cttctcgttg ggcaggttct tatcgaagtt ggtcatccgc
    11221 tcgatgaagc tctgggcgga agcgcccttg tccaccactt cctcgaagtt ccagggggtg
    11281 atggtttcct cgctctttct ggtcatccag gcgaatctgc tgtttcccct ggccagaggg
    11341 cccacgtagt aggggatgcg gaaggtcagg atcttctcga tcttttcccg gttgtccttc
    11401 aggaatgggt aaaaatcttc ctgccgccgc agaatggcgt gcagctctcc caggtggatc
    11461 tggtggggga tgctgccgtt gtcgaaggtc cgctgcttcc gcagcaggtc ctctctgttc
    11521 agcttcacga gcagttcctc ggtgccgtcc atcttttcca ggatgggctt gatgaacttg
    11581 tagaactctt cctggctggc tccgccgtca atgtagccgg cgtagccgtt cttgctctgg
    11641 tcgaagaaaa tctctttgta cttctcaggc agctgctgcc gcacgagagc tttcagcagg
    11701 gtcaggtcct ggtggtgctc gtcgtatctc ttgatcatag aggcgctcag gggggccttg
    11761 gtgatctcgg tgttcactct caggatgtcg ctcagcagga tggcgtcgga caggttcttg
    11821 gcggccagaa acaggtcggc gtactggtcg ccgatctggg ccagcaggtt gtccaggtcg
    11881 tcgtcgtagg tgtccttgct cagctgcagt ttggcatcct cggccaggtc gaagttgctc
    11941 ttgaagttgg gggtcaggcc caggctcagg gcaatcaggt ttccgaacag gccattcttc
    12001 ttctcgccgg gcagctgggc gatcagattt tccagccgtc tgctcttgct cagtctggca
    12061 gacaggatgg ccttggcgtc cacgccgctg gcgttgatgg ggttttcctc gaacagctgg
    12121 ttgtaggtct gcaccagctg gatgaacagc ttgtccacgt cgctgttgtc ggggttcagg
    12181 tcgccctcga tcaggaagtg gccccggaac ttgatcatgt gggccagggc cagatagatc
    12241 agccgcaggt cggccttgtc ggtgctgtcc accagtttct ttctcaggtg gtagatggtg
    12301 gggtacttct cgtggtaggc cacctcgtcc acgatgttgc cgaagatggg gtgccgctcg
    12361 tgcttcttat cctcttccac caggaaggac tcttccagtc tgtggaagaa gctgtcgtcc
    12421 accttggcca tctcgttgct gaagatctct tgcagatagc agatccggtt cttccgtctg
    12481 gtgtatcttc ttctggcggt tctcttcagc cgggtggcct cggctgtttc gccgctgtcg
    12541 aacagcaggg ctccgatcag gttcttcttg atgctgtgcc ggtcggtgtt gcccagcacc
    12601 ttgaatttct tgctgggcac cttgtactcg tcggtgatca cggcccagcc cacagagttg
    12661 gtgccgatgt ccaggccgat gctgtacttc ttgtccatgg tggcagcgct ctagaaccgg
    12721 tcctgtgttc tggcggcaaa cccgttgcga aaaagaacgt tcacggcgac tactgcactt
    12781 atatacggtt ctcccccacc ctcgggaaaa aggcggagcc agtacacgac atcactttcc
    12841 cagtttaccc cgcgccacct tctctaggca ccggatcaat tgccgacccc tccccccaac
    12901 ttctcgggga ctgtgggcga tgtgcgctct gcccactgac gggcaccgga gccaattccc
    12961 actcctttca agacctagct agcgaattca aaaaagcacc gactcggtgc cactttttca
    13021 agttgataac ggactagcct tattttaact tgctatttct agctctaaaa cagagacgta
    13081 caaaaaagag caagaagcta aaaaagattt aaaaattatt tttagcgcag ttaatggaac
    13141 aggaactaaa tttaccccaa aaatattacg tgaatcagga tataacgtta ttgaggttga
    13201 agagcatgca tttgaagatg aaacatttaa aaatgttgta aatccaaatc cagaatttga
    13261 tcctgcatga aaaataccgc ttgaatatgg tattaaacat gatgcagata ttattattat
    13321 gaatgaccca gatgctgaca gatttggaat ggcaataaaa catgatggtc attttgtaag
    13381 attagatgga aatcaaacag gaccaatttt aattgattga aaattatcaa atctaaaacg
    13441 cttaaatagc attccaaaaa atccggctct atattcaagt tttgtaacaa gtgatttggg
    13501 tgatagaatc gctcatgaaa aatatggagt taatattgta aaaactttaa ctggatttaa
    13561 atgaatgggt agagaaattg ctaaagaaga agataacgga ttaaattttg tttttgctta
    13621 tgaagaaagt tatggatatg taattgatga ctcagctaga gataaagatg gaatacaagc
    13681 ttctatatta atagcagagg ctgcttgatt ttataaaaaa caaaataaaa cattagtaga
    13741 ctatttagaa gatttattta aagaaatggg tgcatattac actttcactt taaacttgaa
    13801 ttttaaacca gaagaaaaga aattaaaaat tgaaccatta atgaaatcat tgagagcaac
    13861 acccttaact caaattgctg gacttaaagt tgttaatgtt gaagactaca tcgatggaat
    13921 gtataatatg ccaggacaag acttactaaa attttattta gaagataagt catgatttgc
    13981 tgttcgccca agtggaactg aacctaaact aaaaatttat tttataggtg ttggtgaatc
    14041 tgttcaaaac gctaaagtta aagtagacga aattattaaa gaattaaaat taaaaatgaa
    14101 tatataggag aaaaaatgaa actaaacaaa tatatagatc acacattatt aaaacaagat
    14161 gctacgaaag ctgaaattaa acaattatgt gatgaagcaa ttgaatttga ttttgcaaca
    14221 gtttgtgtta attcatattg aacaagctat tgtaaagaat tattaaaagg cacaaatgta
    14281 ggaataacaa atgttgtagg ttttcctcta ggtgcatgca caacagctac aaaagcattc
    14341 gaagtttctg aagcaattaa agatggtgca acagaaattg atatggtatt aaatattggt
    14401 gcattaaaag acaaaaatta tgaattagtt ttagaagaca tgaaagctgt aaaaaaagca
    14461 gctggatcac atgttgttaa atgtattatg gaaaattgtt tattaacaaa agaagaaatc
    14521 atgaaagctt gtgaaatagc tgttgaagct ggattagaat ttgttaaaac atcaacagga
    14581 ttttcaaaat caggtgcaac atttgaagat gttaaactaa tgaagtcagt tgttaaagac
    14641 aatgctttag ttaaagcagc tggtggagtt agaaca
//
