LOCUS       sequence-209679-       14268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas9 with fluorescent Cherry reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14268)
  AUTHORS   Aubrey BJ, Kelly GL, Kueh AJ, Brennan MS, O'Connor L, Milla L, 
            Wilcox S, Tai L, Strasser A, Herold MJ
  TITLE     An inducible lentiviral guide RNA platform enables the 
            identification of tumor-essential genes and tumor-promoting 
            mutations in vivo.
  JOURNAL   Cell Rep. 2015 Mar 3;10(8):1422-32. doi: 
            10.1016/j.celrep.2015.02.002. Epub 2015 Feb 26.
  PUBMED    25732831
REFERENCE   2  (bases 1 to 14268)
  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.celrep.2015.02.002"; journalName: "Cell Rep"; date: 
            "2015-03-3- 3"; volume: "10"; issue: "8"; pages: "1422-32"
COMMENT     Sequence Label: FUCas9Cherry
FEATURES             Location/Qualifiers
     source          1..14268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(357..1148,1149..1217))
                     /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     980..999
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1218..1322)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1397..1416)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1588..1967
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1969..2167
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2122..2142
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2185..2365
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2412..2537
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    3030..3263
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3448..3492
                     /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    3790..3907
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3972..5183
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5020..5039
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     regulatory      5233..5242
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5242..5307
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5314..5334
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5359..9459
                     /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             5359..9459
                     /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             9460..9507
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9514..9567
                     /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             9568..10278
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(9713..9731)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     9981..10000
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    10306..10894
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(10359..10379)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10777..10788)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10897..10913)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10898..10914)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             11423..11603
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(11629..11646)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11635..11859
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11905..12333
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11992..12011)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12202..12223
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12342..12362)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12347..12676
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12527..12662
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12589..12608
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12724..12771
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12790..13164
                     /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"
     polyA_signal    13294..13415
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13331..13350)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13385..13404
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(13464..13480)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13464..13480)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13477..13499)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13488..13504
                     /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)."
     promoter        complement(join(13512..13518,13519..13536,13537..13542))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13557..13578
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13695..13712)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(13866..14268,1..186))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13946..13965)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
       61 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
      121 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
      181 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
      241 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
      301 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
      361 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
      421 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
      481 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
      541 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
      601 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
      661 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
      721 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
      781 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
      841 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
      901 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
      961 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     1021 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     1081 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     1141 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     1201 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     1261 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     1321 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     1381 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     1441 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     1501 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
     1561 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
     1621 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     1681 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     1741 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     1801 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     1861 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1921 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1981 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     2041 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     2101 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     2161 taagcagcgc gttttgcctg tactgggtct ctctggttag accagatctg agcctgggag
     2221 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     2281 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     2341 tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa gcgaaaggga
     2401 aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg gcaagaggcg
     2461 aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag aaggagagag
     2521 atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg gaaaaaattc
     2581 ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg gcaagcaggg
     2641 agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc tgtagacaaa
     2701 tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga tcattatata
     2761 atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac accaaggaag
     2821 ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag caagcggccg
     2881 ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga attatataaa
     2941 tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa gagaagagtg
     3001 gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt cttgggagca
     3061 gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag acaattattg
     3121 tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca acagcatctg
     3181 ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc tgtggaaaga
     3241 tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact catttgcacc
     3301 actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat ttggaatcac
     3361 acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat acactcctta
     3421 attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga attagataaa
     3481 tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat aaaattattc
     3541 ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact ttctatagtg
     3601 aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc aaccccgagg
     3661 ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag agacagatcc
     3721 attcgattag tgaacggatc ggcactgcgt gcgccaattc tgcagacaaa tggcagtatt
     3781 catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     3841 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
     3901 aaattttcgg gtttattaca gggacagcag agatccagtt tggttaatta acccgtgtcg
     3961 gctccagatc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc ccctcctcac
     4021 ggcgagcgct gccacgtcag acgaagggcg cagcgagcgt cctgatcctt ccgcccggac
     4081 gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa
     4141 ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg
     4201 aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg
     4261 tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc
     4321 gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg
     4381 gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt
     4441 ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt
     4501 tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag
     4561 gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt
     4621 cttgaggcct tcgctaatgc gggaaagctc ttattcgggt gagatgggct ggggcaccat
     4681 ctggggaccc tgacgtgaag tttgtcactg actggagaac tcggtttgtc gtctgttgcg
     4741 ggggcggcag ttatggcggt gccgttgggc agtgcacccg tacctttggg agcgcgcgcc
     4801 ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa tgcagggtgg ggccacctgc
     4861 cggtaggtgt gcggtaggct tttctccgtc gcaggacgca gggttcgggc ctagggtagg
     4921 ctctcctgaa tcgacaggcg ccggacctct ggtgagggga gggataagtg aggcgtcagt
     4981 ttctttggtc ggttttatgt acctatcttc ttaagtagct gaagctccgg ttttgaacta
     5041 tgcgctcggg gttggcgagt gtgttttgtg aagtttttta ggcacctttt gaaatgtaat
     5101 catttgggtc aatatgtaat tttcagtgtt agactagtaa attgtccgct aaattctggc
     5161 cgtttttggc ttttttgtta gacgaagctt gggctgcagg tcgactctag aggatctgaa
     5221 ttctggaccg gtgccaccat ggactataag gaccacgacg gagactacaa ggatcatgat
     5281 attgattaca aagacgatga cgataagatg gccccaaaga agaagcggaa ggtcggtatc
     5341 cacggagtcc cagcagccga caagaagtac agcatcggcc tggacatcgg caccaactct
     5401 gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt caaggtgctg
     5461 ggcaacaccg accggcacag catcaagaag aacctgatcg gagccctgct gttcgacagc
     5521 ggcgaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata caccagacgg
     5581 aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa ggtggacgac
     5641 agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa gcacgagcgg
     5701 caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta ccccaccatc
     5761 taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg gctgatctat
     5821 ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg cgacctgaac
     5881 cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta caaccagctg
     5941 ttcgaggaaa accccatcaa cgccagcggc gtggacgcca aggccatcct gtctgccaga
     6001 ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga gaagaagaat
     6061 ggcctgttcg gaaacctgat tgccctgagc ctgggcctga cccccaactt caagagcaac
     6121 ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga cgacgacctg
     6181 gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc cgccaagaac
     6241 ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat caccaaggcc
     6301 cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct gaccctgctg
     6361 aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt cgaccagagc
     6421 aagaacggct acgccggcta cattgacggc ggagccagcc aggaagagtt ctacaagttc
     6481 atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa gctgaacaga
     6541 gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca ccagatccac
     6601 ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt cctgaaggac
     6661 aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt gggccctctg
     6721 gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac catcaccccc
     6781 tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat cgagcggatg
     6841 accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag cctgctgtac
     6901 gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga gggaatgaga
     6961 aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct gttcaagacc
     7021 aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat cgagtgcttc
     7081 gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg cacataccac
     7141 gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa cgaggacatt
     7201 ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat cgaggaacgg
     7261 ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa gcggcggaga
     7321 tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga caagcagtcc
     7381 ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag
     7441 ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca ggtgtccggc
     7501 cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag
     7561 ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag
     7621 cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag
     7681 aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc
     7741 ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac
     7801 ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac
     7861 tacgatgtgg accatatcgt gcctcagagc tttctgaagg acgactccat cgacaacaag
     7921 gtgctgacca gaagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc
     7981 gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga
     8041 aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc
     8101 ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg
     8161 gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg
     8221 atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg
     8281 cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc
     8341 gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg
     8401 tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag
     8461 tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc
     8521 gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat
     8581 aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg
     8641 aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac
     8701 agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgac
     8761 agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag
     8821 aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag
     8881 aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc
     8941 atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc
     9001 tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc
     9061 ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa
     9121 cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca gatcagcgag
     9181 ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc cgcctacaac
     9241 aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct gtttaccctg
     9301 accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga ccggaagagg
     9361 tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg
     9421 tacgagacac ggatcgacct gtctcagctg ggaggcgaca aaaggccggc ggccacgaaa
     9481 aaggccggcc aggcaaaaaa gaaaaagggc ggtgagggca gaggaagtct gctaacatgc
     9541 ggtgacgtcg aggagaatcc tggcccaatg gtgagcaagg gcgaggagga taacatggcc
     9601 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     9661 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     9721 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     9781 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     9841 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     9901 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     9961 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
    10021 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
    10081 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc
    10141 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag
    10201 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg
    10261 gacgagctgt acaagtagtg acaattcgat atcaagctta tcgataatca acctctggat
    10321 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
    10381 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
    10441 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
    10501 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
    10561 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
    10621 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
    10681 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
    10741 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
    10801 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
    10861 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc tcgatcgaga
    10921 cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc
    10981 ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt
    11041 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg
    11101 actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca
    11161 cacacaaggc tacttccctg attggcagaa ctacacacca gggccaggga tcagatatcc
    11221 actgaccttt ggatggtgct acaagctagt accagttgag caagagaagg tagaagaagc
    11281 caatgaagga gagaacaccc gcttgttaca ccctgtgagc ctgcatggga tggatgaccc
    11341 ggagagagaa gtattagagt ggaggtttga cagccgccta gcatttcatc acatggcccg
    11401 agagctgcat ccggactgta ctgggtctct ctggttagac cagatctgag cctgggagct
    11461 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
    11521 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
    11581 gtcagtgtgg aaaatctcta gcagggcccg tttaaacccg ctgatcagcc tcgactgtgc
    11641 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
    11701 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
    11761 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
    11821 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
    11881 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
    11941 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
    12001 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
    12061 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
    12121 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
    12181 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
    12241 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
    12301 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
    12361 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
    12421 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
    12481 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
    12541 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
    12601 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
    12661 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcagca
    12721 cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg
    12781 aactaaacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc
    12841 ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt ggaggacgac
    12901 ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg
    12961 gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag
    13021 tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat gaccgagatc
    13081 ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac
    13141 ttcgtggccg aggagcagga ctgacacgtg ctacgagatt tcgattccac cgccgccttc
    13201 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
    13261 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
    13321 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
    13381 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
    13441 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
    13501 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
    13561 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
    13621 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
    13681 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
    13741 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
    13801 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
    13861 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
    13921 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
    13981 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
    14041 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
    14101 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
    14161 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
    14221 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagag
//
