LOCUS       sequence-205781-       12311 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SaCas9 expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12311)
  AUTHORS   Boettcher M, Tian R, Blau JA, Markegard E, Wagner RT, Wu D, Mo X, 
            Biton A, Zaitlen N, Fu H, McCormick F, Kampmann M, McManus MT
  TITLE     Dual gene activation and knockout screen reveals directional 
            dependencies in genetic networks.
  JOURNAL   Nat Biotechnol. 2018 Jan 15. pii: nbt.4062. doi: 10.1038/nbt.4062.
  PUBMED    29334369
REFERENCE   2  (bases 1 to 12311)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2018 Jan 15. pii: nbt.4062. doi: 10.1038/nbt.4062."
COMMENT     Sequence Label: pMB-SaCas9
FEATURES             Location/Qualifiers
     source          1..12311
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1684..1917
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2102..2146
                     /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    2444..2561
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     polyA_signal    2749..2883
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2786..2805)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4295..4348)
                     /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             complement(4358..5380)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(4358..5380)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     primer_bind     complement(5457..5481)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(5478..5516)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        5561..5772
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5790..5799
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5802..5822
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5847..9002
                     /codon_start=1
                     /product="Cas9 endonuclease from the Staphylococcus aureus 
                     Type II CRISPR/Cas system"
                     /label=SaCas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="KRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEG
                     RRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEF
                     SAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEV
                     RGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGW
                     KDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKFQ
                     IIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKEI
                     IENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAI
                     NLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIK
                     VINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENAK
                     YLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVKQ
                     EENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFS
                     VQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERN
                     KGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIF
                     ITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLNGLYDKDN
                     DKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKD
                     NGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNL
                     DVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLLN
                     RIEVNMIDITYREYLENMNDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQI
                     IKKG"
     CDS             9003..9050
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9057..9083
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(9111..9144)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    9200..9788
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9253..9273)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9671..9682)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9791..9807)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9792..9808)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10317..10497
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(10559..11147)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10639..10658)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11318..12109,12110..12178))
                     /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     11941..11960
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12179..12283)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga
     1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
     2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
     2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     2581 gcagagatcc agtttggtta gtaccgggcc ctagagatca cgagactagc ctcgagagat
     2641 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     2701 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     2761 gcagcttata atggttacaa ataaggcaat agcatcacaa atttcacaaa taaggcattt
     2821 ttttcactgc attctagttt tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     2881 atctcaaatc cctcggaagc tgcgcctgtc atcgaattcc tgcagcccgg tgcatgacta
     2941 agctagtacc ggttaggatg catgctagct cagttagcct cccccatctc tcgacgcggc
     3001 cgcttcactg gcgctgttgg tgccggcacc tgacacagca gaggatgcag agcccaagga
     3061 aacccagaaa gccgaaggag ccacccagca cgcaagccag gaacactgtc tggttcaccc
     3121 ctctggataa aacctggatc ctggagtcca tcttgacctt atcaccttca ctcagtagac
     3181 actgccacag ccctgtctca ggggccacca cttgaactac tttctgctcc tcagagaccc
     3241 tggcctcctg gttctcctgc ttcagggtca gtctcatctt gggagaggta ggtcccatca
     3301 cctcacaggt caaagtattg ttgagctgag ccactttcat caccaccagg ttcacttcct
     3361 gatgcagtgt ccctttgtcc agagtcagag tcaggttgcc agaaccagca aactgaagcg
     3421 agacctgggg tatcttgagg gtgagtggga gcgtttcctt cagctggagc ttgaggtctt
     3481 tggtggactt ttgtacggac acctctttgt tctttatgga gaaggagatc cagggctgga
     3541 agaaagaatc cttctctgcc ttccacatca gctctcccca cccgttttcc tctgcaaagt
     3601 tgagtgggaa ggagaactcc gctgactctc cctcactctt ataggccgtg atagctgtgc
     3661 tctgaaaacc cagcactgag agtgtcatgc cgaaccagtt ctttttctgg tccagggtca
     3721 cggtgcagtt ccagaagtcg ctgtcctgaa cccttaggtt ggacatggag agaactttgg
     3781 aaccactgac aactttaccc tttttgtgtt tgcactctgt caaggggtta gagaccttag
     3841 agttgctatc caaggtcagg gtcaggctct gcccttgcaa caggctggta cccggactga
     3901 aggtcacttt gaacacccac aactccacct cctctttcct gttctccagc tcacagatat
     3961 aagtctgaga gtcttccatc ttaagtttat tgatgatgag aggaaacgat cctttctccc
     4021 atgccccttt tttggaatca aaacgatcaa actgcgaagg cgaacctcct ctaattaata
     4081 cacctttgcc atgctgcccc agaatcttcc tctggtcaga gaacttccag gtgaagactg
     4141 tgatcttctt ctgggaactc tcgcagggca gttctgctga ttccccttcc ttccccagca
     4201 ccagcgtctt cccttgagtg acagctagga gttgtgacag ctgcagcagc agcagcagca
     4261 agcgcctaag agagatggct cggcacatgt ccggtgggcc aggattctcc tcgacgtcac
     4321 cgcatgttag cagacttcct ctgccctcgg cgcgcccttc ctttgccctc ggacgagtgc
     4381 tggggcgtcg gtttccacta tcggcgagta cttctacaca gccatcggtc cagacggccg
     4441 cgcttctgcg ggcgatttgt gtacgcccga cagtcccggc tccggatcgg acgattgcgt
     4501 cgcatcgacc ctgcgcccaa gctgcatcat cgaaattgcc gtcaaccaag ctctgataga
     4561 gttggtcaag accaatgcgg agcatatacg cccggagccg cggcgatcct gcaagctccg
     4621 gatgcctccg ctcgaagtag cgcgtctgct gctccataca agccaaccac ggcctccaga
     4681 agaagatgtt ggcgacctcg tattgggaat ccccgaacat cgcctcgctc cagtcaatga
     4741 ccgctgttat gcggccattg tccgtcagga cattgttgga gccgaaatcc gcgtgcacga
     4801 ggtgccggac ttcggggcag tcctcggccc aaagcatcag ctcatcgaga gcctgcgcga
     4861 cggacgcact gacggtgtcg tccatcacag tttgccagtg atacacatgg ggatcagcaa
     4921 tcgcgcatat gaaatcacgc catgtagtgt attgaccgat tccttgcggt ccgaatgggc
     4981 cgaacccgct cgtctggcta agatcggccg cagcgatcgc atccatggcc tccgcgaccg
     5041 gctgcagaac agcgggcagt tcggtttcag gcaggtcttg caacgtgaca ccctgtgcac
     5101 ggcgggagat gcaataggtc aggctctcgc tgaattcccc aatgtcaagc acttccggaa
     5161 tcgggagcgc ggccgatgca aagtgccgat aaacataacg atctttgtag aaaccatcgg
     5221 cgcagctatt tacccgcagg acatatccac gccctcctac atcgaagctg aaagcacgag
     5281 attcttcgcc ctccgagagc tgcatcaggt cggagacgct gtcgaacttt tcgatcagaa
     5341 acttctcgac agacgtcgcg gtgagttcag gctttttcat ggtggctcga gcggaggctg
     5401 gatcggtccc ggtgtcttct atggaggtca aaacagcgtg gatggcgtct ccaggcgatc
     5461 tgacggttca ctaaacgagc tctgcttata taggcctccc accgtacacg cctaccctcg
     5521 agaagcttga tatctctaga gctctagtgt gcccgtcagt gggcagagcg cacatcgccc
     5581 acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg
     5641 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
     5701 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc
     5761 gccagaacag tgagctagct ctaccggtcg ccaccatggc cccaaagaag aagcggaagg
     5821 tcggtatcca cggagtccca gcagccaagc ggaactacat cctgggcctg gacatcggca
     5881 tcaccagcgt gggctacggc atcatcgact acgagacacg ggacgtgatc gatgccggcg
     5941 tgcggctgtt caaagaggcc aacgtggaaa acaacgaggg caggcggagc aagagaggcg
     6001 ccagaaggct gaagcggcgg aggcggcata gaatccagag agtgaagaag ctgctgttcg
     6061 actacaacct gctgaccgac cacagcgagc tgagcggcat caacccctac gaggccagag
     6121 tgaagggcct gagccagaag ctgagcgagg aagagttctc tgccgccctg ctgcacctgg
     6181 ccaagagaag aggcgtgcac aacgtgaacg aggtggaaga ggacaccggc aacgagctgt
     6241 ccaccaaaga gcagatcagc cggaacagca aggccctgga agagaaatac gtggccgaac
     6301 tgcagctgga acggctgaag aaagacggcg aagtgcgggg cagcatcaac agattcaaga
     6361 ccagcgacta cgtgaaagaa gccaaacagc tgctgaaggt gcagaaggcc taccaccagc
     6421 tggaccagag cttcatcgac acctacatcg acctgctgga aacccggcgg acctactatg
     6481 agggacctgg cgagggcagc cccttcggct ggaaggacat caaagaatgg tacgagatgc
     6541 tgatgggcca ctgcacctac ttccccgagg aactgcggag cgtgaagtac gcctacaacg
     6601 ccgacctgta caacgccctg aacgacctga acaatctcgt gatcaccagg gacgagaacg
     6661 agaagctgga atattacgag aagttccaga tcatcgagaa cgtgttcaag cagaagaaga
     6721 agcccaccct gaagcagatc gccaaagaaa tcctcgtgaa cgaagaggat attaagggct
     6781 acagagtgac cagcaccggc aagcccgagt tcaccaacct gaaggtgtac cacgacatca
     6841 aggacattac cgcccggaaa gagattattg agaacgccga gctgctggat cagattgcca
     6901 agatcctgac catctaccag agcagcgagg acatccagga agaactgacc aatctgaact
     6961 ccgagctgac ccaggaagag atcgagcaga tctctaatct gaagggctat accggcaccc
     7021 acaacctgag cctgaaggcc atcaacctga tcctggacga gctgtggcac accaacgaca
     7081 accagatcgc tatcttcaac cggctgaagc tggtgcccaa gaaggtggac ctgtcccagc
     7141 agaaagagat ccccaccacc ctggtggacg acttcatcct gagccccgtc gtgaagagaa
     7201 gcttcatcca gagcatcaaa gtgatcaacg ccatcatcaa gaagtacggc ctgcccaacg
     7261 acatcattat cgagctggcc cgcgagaaga actccaagga cgcccagaaa atgatcaacg
     7321 agatgcagaa gcggaaccgg cagaccaacg agcggatcga ggaaatcatc cggaccaccg
     7381 gcaaagagaa cgccaagtac ctgatcgaga agatcaagct gcacgacatg caggaaggca
     7441 agtgcctgta cagcctggaa gccatccctc tggaagatct gctgaacaac cccttcaact
     7501 atgaggtgga ccacatcatc cccagaagcg tgtccttcga caacagcttc aacaacaagg
     7561 tgctcgtgaa gcaggaagaa aacagcaaga agggcaaccg gaccccattc cagtacctga
     7621 gcagcagcga cagcaagatc agctacgaaa ccttcaagaa gcacatcctg aatctggcca
     7681 agggcaaggg cagaatcagc aagaccaaga aagagtatct gctggaagaa cgggacatca
     7741 acaggttctc cgtgcagaaa gacttcatca accggaacct ggtggatacc agatacgcca
     7801 ccagaggcct gatgaacctg ctgcggagct acttcagagt gaacaacctg gacgtgaaag
     7861 tgaagtccat caatggcggc ttcaccagct ttctgcggcg gaagtggaag tttaagaaag
     7921 agcggaacaa ggggtacaag caccacgccg aggacgccct gatcattgcc aacgccgatt
     7981 tcatcttcaa agagtggaag aaactggaca aggccaaaaa agtgatggaa aaccagatgt
     8041 tcgaggaaaa gcaggccgag agcatgcccg agatcgaaac cgagcaggag tacaaagaga
     8101 tcttcatcac cccccaccag atcaagcaca ttaaggactt caaggactac aagtacagcc
     8161 accgggtgga caagaagcct aatagagagc tgattaacga caccctgtac tccacccgga
     8221 aggacgacaa gggcaacacc ctgatcgtga acaatctgaa cggcctgtac gacaaggaca
     8281 atgacaagct gaaaaagctg atcaacaaga gccccgaaaa gctgctgatg taccaccacg
     8341 acccccagac ctaccagaaa ctgaagctga ttatggaaca gtacggcgac gagaagaatc
     8401 ccctgtacaa gtactacgag gaaaccggga actacctgac caagtactcc aaaaaggaca
     8461 acggccccgt gatcaagaag attaagtatt acggcaacaa actgaacgcc catctggaca
     8521 tcaccgacga ctaccccaac agcagaaaca aggtcgtgaa gctgtccctg aagccctaca
     8581 gattcgacgt gtacctggac aatggcgtgt acaagttcgt gaccgtgaag aatctggatg
     8641 tgatcaaaaa agaaaactac tacgaagtga atagcaagtg ctatgaggaa gctaagaagc
     8701 tgaagaagat cagcaaccag gccgagttta tcgcctcctt ctacaacaac gatctgatca
     8761 agatcaacgg cgagctgtat agagtgatcg gcgtgaacaa cgacctgctg aaccggatcg
     8821 aagtgaacat gatcgacatc acctaccgcg agtacctgga aaacatgaac gacaagaggc
     8881 cccccaggat cattaagaca atcgcctcca agacccagag cattaagaag tacagcacag
     8941 acattctggg caacctgtat gaagtgaaat ctaagaagca ccctcagatc atcaaaaagg
     9001 gcaaaaggcc ggcggccacg aaaaaggccg gccaggcaaa aaagaaaaag ggatcctacc
     9061 catacgatgt tccagattac gcttaaatta agaattcgtc gagggaccta ataacttcgt
     9121 atagcataca ttatacgaag ttatacatgt ttaagggttc cggttccact aggtacaatt
     9181 cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     9241 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     9301 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     9361 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     9421 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     9481 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     9541 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     9601 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     9661 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     9721 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     9781 ctccccgcat cgataccgtc gacctcgatc gagacctaga aaaacatgga gcaatcacaa
     9841 gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg
     9901 aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg
     9961 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
    10021 gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc cctgattggc
    10081 agaactacac accagggcca gggatcagat atccactgac ctttggatgg tgctacaagc
    10141 tagtaccagt tgagcaagag aaggtagaag aagccaatga aggagagaac acccgcttgt
    10201 tacaccctgt gagcctgcat gggatggatg acccggagag agaagtatta gagtggaggt
    10261 ttgacagccg cctagcattt catcacatgg cccgagagct gcatccggac tgtactgggt
    10321 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
    10381 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
    10441 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagca
    10501 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
    10561 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
    10621 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
    10681 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
    10741 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
    10801 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
    10861 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
    10921 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
    10981 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
    11041 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
    11101 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
    11161 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
    11221 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
    11281 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
    11341 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
    11401 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
    11461 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
    11521 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
    11581 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
    11641 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
    11701 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
    11761 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
    11821 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
    11881 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
    11941 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
    12001 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
    12061 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
    12121 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
    12181 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
    12241 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
    12301 tgccacctga c
//
