LOCUS       sequence-197194-       11214 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing human codon-optimized S. pyogenes 
            FLAG-tagged Cas9 and blasticidin resistance from EFS promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11214)
  AUTHORS   Hart T, Chandrashekhar M, Aregger M, Steinhart Z, Brown KR, MacLeod 
            G, Mis M, Zimmermann M, Fradet-Turcotte A, Sun S, Mero P, Dirks P, 
            Sidhu S, Roth FP, Rissland OS, Durocher D, Angers S, Moffat J
  TITLE     High-Resolution CRISPR Screens Reveal Fitness Genes and 
            Genotype-Specific Cancer Liabilities.
  JOURNAL   Cell. 2015 Dec 3;163(6):1515-26. doi: 10.1016/j.cell.2015.11.015. 
            Epub 2015 Nov 25.
  PUBMED    26627737
REFERENCE   2  (bases 1 to 11214)
  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"; doi: 
            "10.1016/j.cell.2015.11.015"; journalName: "Cell"; date: "2015-12-3-
            3"; volume: "163"; issue: "6"; pages: "1515-26"
COMMENT     Sequence Label: Lenti‐Cas9‐2A‐Blast
FEATURES             Location/Qualifiers
     source          1..11214
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     66..86
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_feature    287..404
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        473..684
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      704..713
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             713..736
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             743..763
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             788..4888
                     /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             788..4888
                     /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             4889..4936
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4952..5005
                     /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             5006..5404
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    5420..6008
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5473..5493)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5891..5902)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6080..6313
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6391..6512
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6428..6447)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6482..6501
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6552..6687
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6707..6726)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6708..6726)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6736..6753)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6736..6752)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6745..6767)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6894..7349
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6981..7000)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7191..7212
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7375..7479
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7480..7548,7549..8340)
                     /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(7698..7717)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8511..9099
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9000..9019
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9253..9270
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9387..9408
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9423..9428,9429..9446,9447..9453)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9461..9477
                     /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     9466..9488
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9485..9501
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9485..9501
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9520..9540
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9522..9540
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        9568..9794
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             9795..9975
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    10022..10147
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10640..10873
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11058..11102
                     /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"
ORIGIN
        1 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
       61 ccagagaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
      121 gagagataat tagaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
      181 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
      241 atcataaatt ctagatcttg agacaaatgg cagtattcat ccacaatttt aaaagaaaag
      301 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
      361 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
      421 acagcagaga tccactttgg cgccggctcg agtggctccg gtgcccgtca gtgggcagag
      481 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc
      541 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
      601 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
      661 aacgggtttg ccgccagaac acaggtgtcg tgacgcggga tccgccacca tggattacaa
      721 agacgatgac gataagatgg ccccaaagaa gaagcggaag gtcggtatcc acggagtccc
      781 agcagccgac aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc
      841 cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga
      901 ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc
      961 cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat
     1021 ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca
     1081 cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt
     1141 cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag
     1201 aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc
     1261 ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag
     1321 cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa
     1381 ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag
     1441 cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg
     1501 aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc
     1561 cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct
     1621 ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc
     1681 catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc
     1741 ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt
     1801 gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta
     1861 cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat
     1921 cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct
     1981 gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct
     2041 gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa
     2101 gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa
     2161 cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga
     2221 ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga
     2281 taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac
     2341 cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt
     2401 cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt
     2461 gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga
     2521 aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa
     2581 aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat
     2641 cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta
     2701 tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg
     2761 gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat
     2821 cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga
     2881 cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc agggcgatag
     2941 cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg gcatcctgca
     3001 gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc ccgagaacat
     3061 cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga acagccgcga
     3121 gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc tgaaagaaca
     3181 ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc tgcagaatgg
     3241 gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact acgatgtgga
     3301 ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg tgctgaccag
     3361 aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat
     3421 gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa agttcgacaa
     3481 tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct tcatcaagag
     3541 acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg actcccggat
     3601 gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga tcaccctgaa
     3661 gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc gcgagatcaa
     3721 caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa
     3781 aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg
     3841 gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt acttcttcta
     3901 cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg agatccggaa
     3961 gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata agggccggga
     4021 ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga aaaagaccga
     4081 ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca gcgataagct
     4141 gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca gccccaccgt
     4201 ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga aactgaagag
     4261 tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga agaatcccat
     4321 cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca tcaagctgcc
     4381 taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct ctgccggcga
     4441 actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc tgtacctggc
     4501 cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac agctgtttgt
     4561 ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt tctccaagag
     4621 agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca agcaccggga
     4681 taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga ccaatctggg
     4741 agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt acaccagcac
     4801 caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt acgagacacg
     4861 gatcgacctg tctcagctgg gaggcgacaa gcgtcctgct gctactaaga aagctggtca
     4921 agctaagaaa aagaaagcta gcggcagcgg cgagggcaga ggaagtctgc taacatgcgg
     4981 tgatgtcgaa gaaaatccag gcccaatggc caagcctttg tctcaagaag aatccaccct
     5041 cattgaaaga gcaacggcta caatcaacag catccccatc tctgaagact acagcgtcgc
     5101 cagcgcagct ctctctagcg acggccgcat cttcactggt gtcaatgtat atcattttac
     5161 tgggggacct tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg cagctggcaa
     5221 cctgacttgt atcgtcgcga tcggaaatga gaacaggggc atcttgagcc cctgcggacg
     5281 gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa gccatagtga aggacagtga
     5341 tggacagccg acggcagttg ggattcgtga attgctgccc tctggttatg tgtgggaggg
     5401 ctaaacgcgt taagtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     5461 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     5521 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     5581 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     5641 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     5701 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     5761 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     5821 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     5881 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     5941 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     6001 ctccccgcgt cgactttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     6061 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt
     6121 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     6181 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     6241 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     6301 aaaatctcta gcagtacgta tagtagttca tgtcatctta ttattcagta tttataactt
     6361 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     6421 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     6481 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     6541 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     6601 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     6661 agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc
     6721 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     6781 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     6841 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     6901 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     6961 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     7021 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     7081 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     7141 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     7201 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     7261 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     7321 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     7381 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     7441 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     7501 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     7561 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     7621 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     7681 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     7741 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     7801 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     7861 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     7921 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     7981 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     8041 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     8101 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     8161 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     8221 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     8281 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     8341 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     8401 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     8461 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     8521 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     8581 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     8641 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     8701 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     8761 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     8821 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     8881 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     8941 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     9001 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     9061 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     9121 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     9181 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     9241 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     9301 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     9361 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     9421 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     9481 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
     9541 gaacaaaagc tggagctgca agcttaatgt agtcttatgc aatactcttg tagtcttgca
     9601 acatggtaac gatgagttag caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc
     9661 gattggtgga agtaaggtgg tacgatcgtg ccttattagg aaggcaacag acgggtctga
     9721 catggattgg acgaaccact gaattgccgc attgcagaga tattgtattt aagtgcctag
     9781 ctcgatacat aaacgggtct ctctggttag accagatctg agcctgggag ctctctggct
     9841 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     9901 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     9961 ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga aaccagagga
    10021 gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg
    10081 actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag atgggtgcga
    10141 gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc
    10201 agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg agctagaacg
    10261 attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa tactgggaca
    10321 gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata atacagtagc
    10381 aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag ctttagacaa
    10441 gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg ctgatcttca
    10501 gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa tataaagtag
    10561 taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag
    10621 aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca gcaggaagca
    10681 ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg tctggtatag
    10741 tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg ttgcaactca
    10801 cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg
    10861 atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc actgctgtgc
    10921 cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac acgacctgga
    10981 tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta attgaagaat
    11041 cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa tgggcaagtt
    11101 tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc ataatgatag
    11161 taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg aata
//
