LOCUS       Exported               15888 bp ds-DNA     circular SYN 16-AUG-2021
DEFINITION  Express sgRNA/PTG with rice snoRNA U3 promoter and Cas9 with rice 
            ubiquitin promoter for Agrobacterium-mediated transformation..
ACCESSION   .
VERSION     .
KEYWORDS    pRGEB32
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15888)
  AUTHORS   Xie K, Minkenberg B, Yang Y
  TITLE     Boosting CRISPR/Cas9 multiplex editing capability with the 
            endogenous tRNA-processing system.
  JOURNAL   Proc Natl Acad Sci U S A. 2015 Mar 17;112(11):3570-5. doi: 
            10.1073/pnas.1420294112. Epub 2015 Mar 2.
  PUBMED    25733849
REFERENCE   2  (bases 1 to 15888)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 16, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..15888
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             28..75
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     protein_bind    complement(108..132)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             143..166
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      188..440
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(449..466)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(449..465)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(458..480)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    668..692
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             1993..2622
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             3051..4124
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      4190..4384
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(4620..4642)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4728..4868
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4783..4802
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4883..4900)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5054..5642)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5134..5153)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5729..6523)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    6948..6972
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    7050..7224
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(7265..8290)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     promoter        complement(8357..9033)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     complement(8400..8420)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     protein_bind    9224..9245
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9260..9290
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9298..9314
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     9303..9325
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9322..9338
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9322..9338
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        9356..9736
                     /label=OsU3 promoter
                     /note="Oryza sativa (rice) snRNA U3 promoter"
     misc_RNA        9753..9828
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        9909..11618
                     /gene="RUBQ2"
                     /label=Ubi promoter
                     /note="promoter for the rice (Oryza sativa) polyubiquitin 
                     gene RUBQ2"
     intron          10657..11618
                     /note="intron upstream of the coding sequence for the rice 
                     polyubiquitin gene RUBQ2"
     protein_bind    11629..11653
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             11698..11763
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             11770..11790
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11815..27
                     /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"
ORIGIN
        1 atcgacctgt ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag
       61 gcaaaaaaga aaaagtaaga attcgcggcc gcactcgaga tatctagacc cagctttctt
      121 gtacaaagtg gttgataaca gcgactacaa ggatgacgat gacaaggctt agagctcgaa
      181 tttccccgat cgttcaaaca tttggcaata aagtttctta agattgaatc ctgttgccgg
      241 tcttgcgatg attatcatat aatttctgtt gaattacgtt aagcatgtaa taattaacat
      301 gtaatgcatg acgttattta tgagatgggt ttttatgatt agagtcccgc aattatacat
      361 ttaatacgcg atagaaaaca aaatatagcg cgcaaactag gataaattat cgcgcgcggt
      421 gtcatctatg ttactagatc gggaattcac tggccgtcgt tttacaacgt cgtgactggg
      481 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
      541 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
      601 aatgctagag cagcttgagc ttggatcaga ttgtcgtttc ccgccttcag tttaaactat
      661 cagtgtttga caggatatat tggcgggtaa acctaagaga aaagagcgtt tattagaata
      721 atcggatatt taaaagggcg tgaaaaggtt tatccgttcg tccatttgta tgtgcatgcc
      781 aaccacaggg ttcccctcgg gatcaaagta ctttgatcca acccctccgc tgctatagtg
      841 cagtcggctt ctgacgttca gtgcagccgt cttctgaaaa cgacatgtcg cacaagtcct
      901 aagttacgcg acaggctgcc gccctgccct tttcctggcg ttttcttgtc gcgtgtttta
      961 gtcgcataaa gtagaatact tgcgactaga accggagaca ttacgccatg aacaagagcg
     1021 ccgccgctgg cctgctgggc tatgcccgcg tcagcaccga cgaccaggac ttgaccaacc
     1081 aacgggccga actgcacgcg gccggctgca ccaagctgtt ttccgagaag atcaccggca
     1141 ccaggcgcga ccgcccggag ctggccagga tgcttgacca cctacgccct ggcgacgttg
     1201 tgacagtgac caggctagac cgcctggccc gcagcacccg cgacctactg gacattgccg
     1261 agcgcatcca ggaggccggc gcgggcctgc gtagcctggc agagccgtgg gccgacacca
     1321 ccacgccggc cggccgcatg gtgttgaccg tgttcgccgg cattgccgag ttcgagcgtt
     1381 ccctaatcat cgaccgcacc cggagcgggc gcgaggccgc caaggcccga ggcgtgaagt
     1441 ttggcccccg ccctaccctc accccggcac agatcgcgca cgcccgcgag ctgatcgacc
     1501 aggaaggccg caccgtgaaa gaggcggctg cactgcttgg cgtgcatcgc tcgaccctgt
     1561 accgcgcact tgagcgcagc gaggaagtga cgcccaccga ggccaggcgg cgcggtgcct
     1621 tccgtgagga cgcattgacc gaggccgacg ccctggcggc cgccgagaat gaacgccaag
     1681 aggaacaagc atgaaaccgc accaggacgg ccaggacgaa ccgtttttca ttaccgaaga
     1741 gatcgaggcg gagatgatcg cggccgggta cgtgttcgag ccgcccgcgc acgtctcaac
     1801 cgtgcggctg catgaaatcc tggccggttt gtctgatgcc aagctggcgg cctggccggc
     1861 cagcttggcc gctgaagaaa ccgagcgccg ccgtctaaaa aggtgatgtg tatttgagta
     1921 aaacagcttg cgtcatgcgg tcgctgcgta tatgatgcga tgagtaaata aacaaatacg
     1981 caaggggaac gcatgaaggt tatcgctgta cttaaccaga aaggcgggtc aggcaagacg
     2041 accatcgcaa cccatctagc ccgcgccctg caactcgccg gggccgatgt tctgttagtc
     2101 gattccgatc cccagggcag tgcccgcgat tgggcggccg tgcgggaaga tcaaccgcta
     2161 accgttgtcg gcatcgaccg cccgacgatt gaccgcgacg tgaaggccat cggccggcgc
     2221 gacttcgtag tgatcgacgg agcgccccag gcggcggact tggctgtgtc cgcgatcaag
     2281 gcagccgact tcgtgctgat tccggtgcag ccaagccctt acgacatatg ggccaccgcc
     2341 gacctggtgg agctggttaa gcagcgcatt gaggtcacgg atggaaggct acaagcggcc
     2401 tttgtcgtgt cgcgggcgat caaaggcacg cgcatcggcg gtgaggttgc cgaggcgctg
     2461 gccgggtacg agctgcccat tcttgagtcc cgtatcacgc agcgcgtgag ctacccaggc
     2521 actgccgccg ccggcacaac cgttcttgaa tcagaacccg agggcgacgc tgcccgcgag
     2581 gtccaggcgc tggccgctga aattaaatca aaactcattt gagttaatga ggtaaagaga
     2641 aaatgagcaa aagcacaaac acgctaagtg ccggccgtcc gagcgcacgc agcagcaagg
     2701 ctgcaacgtt ggccagcctg gcagacacgc cagccatgaa gcgggtcaac tttcagttgc
     2761 cggcggagga tcacaccaag ctgaagatgt acgcggtacg ccaaggcaag accattaccg
     2821 agctgctatc tgaatacatc gcgcagctac cagagtaaat gagcaaatga ataaatgagt
     2881 agatgaattt tagcggctaa aggaggcggc atggaaaatc aagaacaacc aggcaccgac
     2941 gccgtggaat gccccatgtg ttgaggaacg ggcggttggc caggcgtaag cggctgggtt
     3001 gcctgccggc cctgcaatgg cactggaacc cccaagcccg aggaatcggc gtgagcggtc
     3061 gcaaaccatc cggcccggta caaatcggcg cggcgctggg tgatgacctg gtggagaagt
     3121 tgaaggccgc gcaggccgcc cagcggcaac gcatcgaggc agaagcacgc cccggtgaat
     3181 cgtggcaagc ggccgctgat cgaatccgca aagaatcccg gcaaccgccg gcagccggtg
     3241 cgccgtcgat taggaagccg cccaagggcg acgagcaacc agattttttc gttccgatgc
     3301 tctatgacgt gggcacccgc gatagtcgca gcatcatgga cgtggccgtt ttccgtctgt
     3361 cgaagcgtga ccgacgagct ggcgaggtga tccgctacga gcttccagac gggcacgtag
     3421 aggtttccgc agggccggcc ggcatggcca gtgtgtggga ttacgacctg gtactgatgg
     3481 cggtttccca tctaaccgaa tccatgaacc gataccggga agggaaggga gacaagcccg
     3541 gccgcgtgtt ccgtccacac gttgcggacg tactcaagtt ctgccggcga gccgatggcg
     3601 gaaagcagaa agacgacctg gtagaaacct gcattcggtt aaacaccacg cacgttgcca
     3661 tgcagcgtac gaagaaggcc aagaacggcc gcctggtgac ggtatccgag ggtgaagcct
     3721 tgattagccg ctacaagatc gtaaagagcg aaaccgggcg gccggagtac atcgagatcg
     3781 agctagctga ttggatgtac cgcgagatca cagaaggcaa gaacccggac gtgctgacgg
     3841 ttcaccccga ttactttttg atcgatcccg gcatcggccg ttttctctac cgcctggcac
     3901 gccgcgccgc aggcaaggca gaagccagat ggttgttcaa gacgatctac gaacgcagtg
     3961 gcagcgccgg agagttcaag aagttctgtt tcaccgtgcg caagctgatc gggtcaaatg
     4021 acctgccgga gtacgatttg aaggaggagg cggggcaggc tggcccgatc ctagtcatgc
     4081 gctaccgcaa cctgatcgag ggcgaagcat ccgccggttc ctaatgtacg gagcagatgc
     4141 tagggcaaat tgccctagca ggggaaaaag gtcgaaaagc actctttcct gtgaatagca
     4201 cgtacattgg gaacccaaag ccgtacattg ggaaccggaa cccgtacatt gggaacccaa
     4261 agccgtacat tgggaaccgg tcacacatgt aagtgactga tataaaagag aaaaaaggcg
     4321 atttttccgc ctaaaactct ttaaaactta ttaaaactct taaaacccgc ctggcctgtg
     4381 cataactgtc tggccagcgc acagccgaag agctgcaaaa agcgcctacc cttcggtcgc
     4441 tgcgctccct acgccccgcc gcttcgcgtc ggcctatcgc ggccgctggc cgctcaaaaa
     4501 tggctggcct acggccaggc aatctaccag ggcgcggaca agccgcgccg tcgccactcg
     4561 accgccggcg cccacatcaa ggcaccctgc ctcgcgcgtt tcggtgatga cggtgaaaac
     4621 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     4681 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc
     4741 cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg
     4801 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     4861 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4921 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4981 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5041 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5101 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5161 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5221 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5281 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5341 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5401 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5461 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     5521 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5581 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5641 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5701 aagggatttt ggtcatgcat tctaggtact aaaacaattc atccagtaaa atataatatt
     5761 ttattttctc ccaatcaggc ttgatcccca gtaagtcaaa aaatagctcg acatactgtt
     5821 cttccccgat atcctccctg atcgaccgga cgcagaaggc aatgtcatac cacttgtccg
     5881 ccctgccgct tctcccaaga tcaataaagc cacttacttt gccatctttc acaaagatgt
     5941 tgctgtctcc caggtcgccg tgggaaaaga caagttcctc ttcgggcttt tccgtcttta
     6001 aaaaatcata cagctcgcgc ggatctttaa atggagtgtc ttcttcccag ttttcgcaat
     6061 ccacatcggc cagatcgtta ttcagtaagt aatccaattc ggctaagcgg ctgtctaagc
     6121 tattcgtata gggacaatcc gatatgtcga tggagtgaaa gagcctgatg cactccgcat
     6181 acagctcgat aatcttttca gggctttgtt catcttcata ctcttccgag caaaggacgc
     6241 catcggcctc actcatgagc agattgctcc agccatcatg ccgttcaaag tgcaggacct
     6301 ttggaacagg cagctttcct tccagccata gcatcatgtc cttttcccgt tccacatcat
     6361 aggtggtccc tttataccgg ctgtccgtca tttttaaata taggttttca ttttctccca
     6421 ccagcttata taccttagca ggagacattc cttccgtatc ttttacgcag cggtattttt
     6481 cgatcagttt tttcaattcc ggtgatattc tcattttagc catttattat ttccttcctc
     6541 ttttctacag tatttaaaga taccccaaga agctaattat aacaagacga actccaattc
     6601 actgttcctt gcattctaaa accttaaata ccagaaaaca gctttttcaa agttgttttc
     6661 aaagttggcg tataacatag tatcgacgga gccgattttg aaaccgcggt gatcacaggc
     6721 agcaacgctc tgtcatcgtt acaatcaaca tgctaccctc cgcgagatca tccgtgtttc
     6781 aaacccggca gcttagttgc cgttcttccg aatagcatcg gtaacatgag caaagtctgc
     6841 cgccttacaa cggctctccc gctgacgccg tcccggactg atgggctgcc tgtatcgagt
     6901 ggtgattttg tgccgagctg ccggtcgggg agctgttggc tggctggtgg caggatatat
     6961 tgtggtgtaa acaaattgac gcttagacaa cttaataaca cattgcggac gtttttaatg
     7021 tactgaatta acgccgaatt aattcggggg atctggattt tagtactgga ttttggtttt
     7081 aggaattaga aattttattg atagaagtat tttacaaata caaatacata ctaagggttt
     7141 cttatatgct caacacatga gcgaaaccct ataggaaccc taattccctt atctgggaac
     7201 tactcacaca ttattatgga gaaactcgag cttgtcgatc gacagatccc ggtcggcatc
     7261 tactctattt ctttgccctc ggacgagtgc tggggcgtcg gtttccacta tcggcgagta
     7321 cttctacaca gccatcggtc cagacggccg cgcttctgcg ggcgatttgt gtacgcccga
     7381 cagtcccggc tccggatcgg acgattgcgt cgcatcgacc ctgcgcccaa gctgcatcat
     7441 cgaaattgcc gtcaaccaag ctctgataga gttggtcaag accaatgcgg agcatatacg
     7501 cccggagtcg tggcgatcct gcaagctccg gatgcctccg ctcgaagtag cgcgtctgct
     7561 gctccataca agccaaccac ggcctccaga agaagatgtt ggcgacctcg tattgggaat
     7621 ccccgaacat cgcctcgctc cagtcaatga ccgctgttat gcggccattg tccgtcagga
     7681 cattgttgga gccgaaatcc gcgtgcacga ggtgccggac ttcggggcag tcctcggccc
     7741 aaagcatcag ctcatcgaga gcctgcgcga cggacgcact gacggtgtcg tccatcacag
     7801 tttgccagtg atacacatgg ggatcagcaa tcgcgcatat gaaatcacgc catgtagtgt
     7861 attgaccgat tccttgcggt ccgaatgggc cgaacccgct cgtctggcta agatcggccg
     7921 cagcgatcgc atccatagcc tccgcgaccg gttgtagaac agcgggcagt tcggtttcag
     7981 gcaggtcttg caacgtgaca ccctgtgaac ggcgggagat gcaataggtc aggctctcgc
     8041 taaactcccc aatgtcaagc acttccggaa tcgggagcgc ggccgatgca aagtgccgat
     8101 aaacataacg atctttgtag aaaccatcgg cgcagctatt tacccgcagg acatatccac
     8161 gccctcctac atcgaagctg aaagcacgag attcttcgcc ctccgagagc tgcatcaggt
     8221 cggagacgct gtcgaacttt tcgatcagaa acttctcgac agacgtcgcg gtgagttcag
     8281 gctttttcat atctcattgc cccccggatc tgcgaaagct cgagagagat agatttgtag
     8341 agagagactg gtgatttcag cgtgtcctct ccaaatgaaa tgaacttcct tatatagagg
     8401 aagggtcttg cgaaggatag tgggattgtg cgtcatccct tacgtcagtg gagatatcac
     8461 atcaatccac ttgctttgaa gacgtggttg gaacgtcttc tttttccacg atgctcctcg
     8521 tgggtggggg tccatctttg ggaccactgt cggcagaggc atcttgaacg atagcctttc
     8581 ctttatcgca atgatggcat ttgtaggtgc caccttcctt ttctactgtc cttttgatga
     8641 agtgacagat agctgggcaa tggaatccga ggaggtttcc cgatattacc ctttgttgaa
     8701 aagtctcaat agccctttgg tcttctgaga ctgtatcttt gatattcttg gagtagacga
     8761 gagtgtcgtg ctccaccatg ttcacatcaa tccacttgct ttgaagacgt ggttggaacg
     8821 tcttcttttt ccacgatgct cctcgtgggt gggggtccat ctttgggacc actgtcggca
     8881 gaggcatctt gaacgatagc ctttccttta tcgcaatgat ggcatttgta ggtgccacct
     8941 tccttttcta ctgtcctttt gatgaagtga cagatagctg ggcaatggaa tccgaggagg
     9001 tttcccgata ttaccctttg ttgaaaagtc tcaatagccc tttggtcttc tgagactgta
     9061 tctttgatat tcttggagta gacgagagtg tcgtgctcca ccatgttggc aagctgctct
     9121 agccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     9181 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     9241 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     9301 tgagcggata acaatttcac acaggaaaca gctatgacat gattacgcca agcttaagga
     9361 atctttaaac atacgaacag atcacttaaa gttcttctga agcaacttaa agttatcagg
     9421 catgcatgga tcttggagga atcagatgtg cagtcaggga ccatagcaca agacaggcgt
     9481 cttctactgg tgctaccagc aaatgctgga agccgggaac actgggtacg ttggaaacca
     9541 cgtgatgtga agaagtaaga taaactgtag gagaaaagca tttcgtagtg ggccatgaag
     9601 cctttcagga catgtattgc agtatgggcc ggcccattac gcaattggac gacaacaaag
     9661 actagtatta gtaccacctc ggctatccac atagatcaaa gctgatttaa aagagttgtg
     9721 cagatgatcc gtggcaggag accgaggtct cggttttaga gctagaaata gcaagttaaa
     9781 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt ttttgtttta
     9841 gagctagaaa tagcaagtta aaataaggct agtccgtttt tagcgcgtgc atgcctgcag
     9901 gtccacaaat tcgggtcaag gcggaagcca gcgcgccacc ccacgtcagc aaatacggag
     9961 gcgcggggtt gacggcgtca cccggtccta acggcgacca acaaaccagc cagaagaaat
    10021 tacagtaaaa aaaaagtaaa ttgcactttg atccaccttt tattacctaa gtctcaattt
    10081 ggatcaccct taaacctatc ttttcaattt gggccgggtt gtggtttgga ctaccatgaa
    10141 caacttttcg tcatgtctaa cttccctttc agcaaacata tgaaccatat atagaggaga
    10201 tcggccgtat actagagctg atgtgtttaa ggtcgttgat tgcacgagaa aaaaaaatcc
    10261 aaatcgcaac aatagcaaat ttatctggtt caaagtgaaa agatatgttt aaaggtagtc
    10321 caaagtaaaa cttatagata ataaaatgtg gtccaaagcg taattcactc aaaaaaaatc
    10381 aacgagacgt gtaccaaacg gagacaaacg gcatcttctc gaaatttccc aaccgctcgc
    10441 tcgcccgcct cgtcttcccg gaaaccgcgg tggtttcagc gtggcggatt ctccaagcag
    10501 acggagacgt cacggcacgg gactcctccc accacccaac cgccataaat accagccccc
    10561 tcatctcctc tcctcgcatc agctccaccc ccgaaaaatt tctccccaat ctcgcgaggc
    10621 tctcgtcgtc gaatcgaatc ctctcgcgtc ctcaaggtac gctgcttctc ctctcctcgc
    10681 ttcgtttcga ttcgatttcg gacgggtgag gttgttttgt tgctagatcc gattggtggt
    10741 tagggttgtc gatgtgatta tcgtgagatg tttaggggtt gtagatctga tggttgtgat
    10801 ttgggcacgg ttggttcgat aggtggaatc gtggttaggt tttgggattg gatgttggtt
    10861 ctgatgattg gggggaattt ttacggttag atgaattgtt ggatgattcg attggggaaa
    10921 tcggtgtaga tctgttgggg aattgtggaa ctagtcatgc ctgagtgatt ggtgcgattt
    10981 gtagcgtgtt ccatcttgta ggccttgttg cgagcatgtt cagatctact gttccgctct
    11041 tgattgagtt attggtgcca tgggttggtg caaacacagg ctttaatatg ttatatctgt
    11101 tttgtgtttg atgtagatct gtagggtagt tcttcttaga catggttcaa ttatgtagct
    11161 tgtgcgtttc gatttgattt catatgttca cagattagat aatgatgaac tcttttaatt
    11221 aattgtcaat ggtaaatagg aagtcttgtc gctatatctg tcataatgat ctcatgttac
    11281 tatctgccag taatttatgc taagaactat attagaatat catgttacaa tctgtagtaa
    11341 tatcatgtta caatctgtag ttcatctata taatctattg tggtaatttc tttttactat
    11401 ctgtgtgaag attattgcca ctagttcatt ctacttattt ctgaagttca ggatacgtgt
    11461 gctgttacta cctatctgaa tacatgtgtg atgtgcctgt tactatcttt ttgaatacat
    11521 gtatgttctg ttggaatatg tttgctgttt gatccgttgt tgtgtcctta atcttgtgct
    11581 agttcttacc ctatctgttt ggtgattatt tcttgcagat agttatcaac aagtttgtac
    11641 aaaaaagcag gcttcgaagg agatagaacc aattctctaa ggaaatactt aaccatggac
    11701 tataaggacc acgacggaga ctacaaggat catgatattg attacaaaga cgatgacgat
    11761 aagatggccc caaagaagaa gcggaaggtc ggtatccacg gagtcccagc agccgacaag
    11821 aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac
    11881 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc
    11941 aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg
    12001 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa
    12061 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag
    12121 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg
    12181 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg
    12241 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag
    12301 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
    12361 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc
    12421 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa
    12481 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc
    12541 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa
    12601 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
    12661 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc
    12721 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag
    12781 agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg
    12841 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt
    12901 gacggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg
    12961 gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg
    13021 accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg
    13081 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc
    13141 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc
    13201 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac
    13261 aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc
    13321 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag
    13381 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag
    13441 cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag
    13501 ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg
    13561 gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac
    13621 aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg
    13681 acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc
    13741 gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc
    13801 cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg
    13861 aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc
    13921 tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac
    13981 attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg
    14041 gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg
    14101 gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg
    14161 atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac
    14221 acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac
    14281 gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct
    14341 cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac
    14401 cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg
    14461 cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc
    14521 gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa
    14581 acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac
    14641 gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg
    14701 tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac
    14761 gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag
    14821 ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc
    14881 aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg
    14941 aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc
    15001 gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg
    15061 cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc
    15121 ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag
    15181 aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg
    15241 ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg
    15301 ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa
    15361 gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg
    15421 ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga
    15481 aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag
    15541 aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag
    15601 cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc
    15661 gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga
    15721 gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc
    15781 ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg
    15841 gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacgg
//
