LOCUS       sequence-249659-       14039 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cas9 expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14039)
  AUTHORS   Wang C, Lu T, Emanuel G, Babcock HP, Zhuang X
  TITLE     Imaging-based pooled CRISPR screening reveals regulators of lncRNA 
            localization.
  JOURNAL   Proc Natl Acad Sci U S A. 2019 May 28;116(22):10842-10851. doi: 
            10.1073/pnas.1903808116. Epub 2019 May 13.
  PUBMED    31085639
REFERENCE   2  (bases 1 to 14039)
  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: 
            "10.1073/pnas.1903808116"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2019-05-28- 28"; volume: "116"; issue: "22"; pages: 
            "10842-10851"
COMMENT     Sequence Label: pFUGW Cas9-BFP
FEATURES             Location/Qualifiers
     source          1..14039
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        618..816
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             834..1014
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1061..1186
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1679..1912
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2097..2141
                     /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    2439..2556
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3283..3690
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      3759..3768
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3765..7868
                     /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             3765..7868
                     /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             7869..7916
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7917..7940
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7947..8642
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     CDS             8658..8678
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8682..8702
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    8730..9318
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8783..8803)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9201..9212)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9321..9337)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9322..9338)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9843..10023
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10049..10066)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10055..10279
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10325..10753
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10412..10431)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10622..10643
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10762..10782)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10767..11096
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10947..11082
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11009..11028
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11144..11191
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11210..11584
                     /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    11714..11835
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11751..11770)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11805..11824
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11884..11900)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11884..11900)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11897..11919)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11908..11924
                     /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(11932..11938,11939..11956,11957..11962))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11977..11998
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12115..12132)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12286..12874)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12366..12385)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13045..13836,13837..13905))
                     /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     13668..13687
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13906..14010)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc
      841 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      901 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      961 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     1021 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     1081 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     1141 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     1201 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     1261 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     1321 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     1381 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     1441 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     1501 aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga
     1561 gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag
     1621 tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag
     1681 gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga
     1741 cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc
     1801 tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc
     1861 tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt
     1921 ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta
     1981 ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta
     2041 acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga
     2101 atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa
     2161 caaattggct gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa
     2221 gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat
     2281 cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag
     2341 aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg gcactgcgtg
     2401 cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt
     2461 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
     2521 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga
     2581 gatccagttt ggttaattaa cgggaggtgg tccctgcagt tacgccaatg ataacccccg
     2641 ccagaaaaat cttagtagcc ttcccttttt gttttccgtg ccccaactcg gcggattgac
     2701 tcggcccctt ccggaaacac ccgaatcaac ttctagtcaa attattgttc acgccgcaat
     2761 gacccacccc tggcccgcgt ctgtggaact gacccctggt gtacaggaga gttcgctgct
     2821 gaaagtggtc ccaaaggggt actagttttt aagctcccaa ctccccctcc cccagcgtct
     2881 ggaggattcc acaccctcgc accgcagggg cgaggaagtg ggcggagtcc ggttttggcg
     2941 ccagccgctg aggctgccaa gcagaaaagc caccgctgag gagactccgg tcactgtcct
     3001 cgccccgcct cccccttccc tccccttggg gaccaccggg cgccacgccg cgaacggtaa
     3061 gtgccgcggt cgtcggcgcc tccgccctcc ccctagggcc ccaattccca gcgggcgcgg
     3121 cgcgcggccc cgcgcgcagc tcccggctcc ctcccccttc ggatgtggct tgagctgtag
     3181 gcgcggagtc ctgcagcccc gataaaataa aagattttat ttagtctcca gaaaaagggg
     3241 ggaatgaaag accccacctg taggtttggc aagctagctg cagtaacgcc attttgcaag
     3301 gcatggaaaa ataccaaacc aagaatagag aagttcagat caagggcggg tacatgaaaa
     3361 tagctaacgt tgggccaaac aggatatctg cggtgagcag tttcggcccc ggcccggggc
     3421 caagaacaga tggtcaccgc agtttcggcc ccggcccgag gccaagaaca gatggtcccc
     3481 agatatggcc caaccctcag cagtttctta agacccatca gatgtttcca ggctccccca
     3541 aggacctgaa atgaccctgc gccttatttg aattaaccaa tcagcctgct tctcgcttct
     3601 gttcgcgcgc ttctgcttcc cgagctctat aaaagagctc acaacccctc actcggcgcg
     3661 ccagtcctcc gacagactga gtcgcccggg ggggatctgg agctctcgag aattctcacg
     3721 cgtctgcagg atatcaagct tgcggtaccg cgggcccggc caccatggac aagaagtaca
     3781 gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca
     3841 aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga
     3901 acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga
     3961 gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg caagagatct
     4021 tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc
     4081 tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg
     4141 tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca
     4201 ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg
     4261 gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca
     4321 tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg
     4381 tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga
     4441 tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc
     4501 tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc
     4561 tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt
     4621 acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc
     4681 tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg
     4741 acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga
     4801 agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg
     4861 gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca
     4921 ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg
     4981 acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc
     5041 aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct
     5101 tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga
     5161 ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg
     5221 cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga
     5281 aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca
     5341 aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa
     5401 aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag
     5461 aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc
     5521 ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact
     5581 tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt
     5641 ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca
     5701 aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc
     5761 tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg
     5821 acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag
     5881 aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca
     5941 atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg
     6001 agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag
     6061 agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag
     6121 agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc
     6181 tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc
     6241 aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct
     6301 ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca
     6361 agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc
     6421 tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag
     6481 gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc
     6541 agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga
     6601 atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt
     6661 tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg
     6721 acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa
     6781 gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg
     6841 agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt
     6901 tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa
     6961 acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag
     7021 tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca
     7081 gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact
     7141 gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg
     7201 tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctactgggga
     7261 tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg
     7321 gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc
     7381 tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac
     7441 tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga
     7501 agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc
     7561 tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta
     7621 atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg
     7681 ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt
     7741 actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca
     7801 ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg
     7861 gaggcgacaa gcgacctgcc gccacaaaga aggctggaca ggctaagaag aagaaagatt
     7921 acaaagacga tgacgataag ggatccagcg agctgattaa ggagaacatg cacatgaagc
     7981 tgtacatgga gggcaccgtg gacaaccatc acttcaagtg cacatccgag ggcgaaggca
     8041 agccctacga gggcacccag accatgagaa tcaaggtggt cgagggcggc cctctcccct
     8101 tcgccttcga catcctggct actagcttcc tctacggcag caagaccttc atcaaccaca
     8161 cccagggcat ccccgacttc ttcaagcagt ccttccctga gggcttcaca tgggagagag
     8221 tcaccacata cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg
     8281 gctgcctcat ctacaacgtc aagatcagag gggtgaactt cacatccaac ggccctgtga
     8341 tgcagaagaa aacactcggc tgggaggcct tcaccgagac gctgtacccc gctgacggcg
     8401 gcctggaagg cagaaacgac atggccctga agctcgtggg cgggagccat ctgatcgcaa
     8461 acatcaagac cacatataga tccaagaaac ccgctaagaa cctcaagatg cctggcgtct
     8521 actatgtgga ctacagactg gaaagaatca aggaggccaa caacgagacc tacgtcgagc
     8581 agcacgaggt ggcagtggcc agatactgcg acctccctag caaactgggg cacaagctta
     8641 atggtggtgg tactagtccc aagaagaagc gcaaggtgga tccaaaaaag aagagaaagg
     8701 tataagaatt cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     8761 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     8821 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     8881 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     8941 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     9001 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     9061 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     9121 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     9181 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     9241 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     9301 tttgggccgc ctccccgcat cgataccgtc gacctcgaga cctagaaaaa catggagcaa
     9361 tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa gcacaagagg
     9421 aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg acttacaagg
     9481 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     9541 cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc tacttccctg
     9601 attggcagaa ctacacacca gggccaggga tcagatatcc actgaccttt ggatggtgct
     9661 acaagctagt accagttgag caagagaagg tagaagaagc caatgaagga gagaacaccc
     9721 gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa gtattagagt
     9781 ggaggtttga cagccgccta gcatttcatc acatggcccg agagctgcat ccggactgta
     9841 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     9901 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     9961 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
    10021 gcagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
    10081 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
    10141 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
    10201 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
    10261 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
    10321 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
    10381 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
    10441 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
    10501 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
    10561 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
    10621 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
    10681 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
    10741 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
    10801 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
    10861 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
    10921 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
    10981 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
    11041 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
    11101 tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca attaatcatc
    11161 ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca tggccaagtt
    11221 gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac
    11281 cgaccggctc gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga
    11341 cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc
    11401 ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac
    11461 gaacttccgg gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg
    11521 ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga
    11581 ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
    11641 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
    11701 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
    11761 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
    11821 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
    11881 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
    11941 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
    12001 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
    12061 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
    12121 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
    12181 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
    12241 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
    12301 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
    12361 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
    12421 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
    12481 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
    12541 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
    12601 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
    12661 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
    12721 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
    12781 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
    12841 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
    12901 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
    12961 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
    13021 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
    13081 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
    13141 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
    13201 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
    13261 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
    13321 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
    13381 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
    13441 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
    13501 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
    13561 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
    13621 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
    13681 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
    13741 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
    13801 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
    13861 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
    13921 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
    13981 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacg
//
