LOCUS       Exported               16544 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  CRISPR/Cas9-mediated genome editing in Arabidopsis. Contains Cas9 
            and empty gRNA scaffold, Kan resistance.
ACCESSION   .
VERSION     .
KEYWORDS    pKEE401
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16544)
  AUTHORS   Wang ZP, Xing HL, Dong L, Zhang HY, Han CY, Wang XC, Chen QJ
  TITLE     Egg cell-specific promoter-controlled CRISPR/Cas9 efficiently 
            generates homozygous mutants for multiple target genes in 
            Arabidopsis in a single generation.
  JOURNAL   Genome Biol. 2015 Jul 21;16:144. doi: 10.1186/s13059-015-0715-0.
  PUBMED    26193878
REFERENCE   2  (bases 1 to 16544)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..16544
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..25
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     213..235
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     227..244
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     228..244
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             957..1748
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     misc_RNA        1900..1975
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             3193..3258
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3265..3285
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3310..7410
                     /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"
     RBS             7323..7331
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             7411..7458
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(8115..8131)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8115..8131)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8128..8150)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8139..8155
                     /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(8163..8193)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8208..8229
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8420..9096
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     9033..9053
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             9159..9956
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(9216..9235)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9826..9845
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    10013..10187
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    10265..10289
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             10714..11508
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      11595..12183
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12084..12103
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12337..12354
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    12369..12509
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(12435..12454)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12595..12617
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      12853..13047
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(13113..14186)
                     /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"
     CDS             complement(14615..15244)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 gtttacccgc caatatatcc tgtcaaacac tgatagttta aactgaaggc gggaaacgac
       61 aatctgatcc aagctcaagc tgctctagca ttcgccattc aggctgcgca actgttggga
      121 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
      181 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
      241 cagtgccaag cttcgacttg ccttccgcac aatacatcat ttcttcttag ctttttttct
      301 tcttcttcgt tcatacagtt tttttttgtt tatcagctta cattttcttg aaccgtagct
      361 ttcgttttct tctttttaac tttccattcg gagtttttgt atcttgtttc atagtttgtc
      421 ccaggattag aatgattagg catcgaacct tcaagaattt gattgaataa aacatcttca
      481 ttcttaagat atgaagataa tcttcaaaag gcccctggga atctgaaaga agagaagcag
      541 gcccatttat atgggaaaga acaatagtat ttcttatata ggcccattta agttgaaaac
      601 aatcttcaaa agtcccacat cgcttagata agaaaacgaa gctgagttta tatacagcta
      661 gagtcgaagt agtgattggg agaccaaccc agtggacata agcctgttcg gttcgtaagc
      721 tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
      781 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt ggggtacagt
      841 ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat
      901 ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta aacatcatgg
      961 gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc gtcatcgagc
     1021 gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc
     1081 tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa
     1141 cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagcgaga
     1201 ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc
     1261 cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gcaggtatct
     1321 tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata
     1381 gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc
     1441 tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg
     1501 atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa
     1561 tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc
     1621 ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc ttggcctcgc
     1681 gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc aaggtagtcg
     1741 gcaaataatg tctagctaga aattcgttca agccgacgcc gcttcgcggc gcggcttaac
     1801 tcaagcgtta gatgcactaa gcacataatt gctcacagcc aaactatcag gtcaagtctg
     1861 cttttattat ttttaagcgt gcataataag ccggtctcgg ttttagagct agaaatagca
     1921 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
     1981 tttgcaaaat tttccagatc gatttcttct tcctctgttc ttcggcgttc aatttctggg
     2041 gttttctctt cgttttctgt aactgaaacc taaaatttga cctaaaaaaa atctcaaata
     2101 atatgattca gtggttttgt acttttcagt tagttgagtt ttgcagttcc gatgagataa
     2161 accaatacca tggactttga taaatgttcc tcgctgacgt aagaagacat tagtaatggt
     2221 tataatatat agctttctat gaatgtatgg tgagaaaatg tctgttcact gattttgagt
     2281 ttggaataaa agcatttgcg tttggtttat cattgcgttt atacaaggac agagatccac
     2341 tgagctggaa tagcttaaaa ccattatcag aacaaaataa accatttttt gttaagaatc
     2401 agagcatagt aaacaacaga aacaacctaa gagaggtaac ttgtccaaga agatagctaa
     2461 ttatatctat tttataaaag ttatcatagt ttgtaagtca caaaagatgc aaataacaga
     2521 gaaactagga gacttgagaa tatacattct tgtatatttg tattcgagat tgtgaaaatt
     2581 tgaccataag tttaaattct taaaaagata tatctgatct agatgatggt tatagactgt
     2641 aattttacca catgtttaat gatggatagt gacacacatg acacatcgac aacactatag
     2701 catcttattt agattacaac atgaaatttt tctgtaatac atgtctttgt acataattta
     2761 aaagtaattc ctaagaaata tatttataca aggagtttaa agaaaacata gcataaagtt
     2821 caatgagtag taaaaaccat atacagtata tagcataaag ttcaatgagt ttattacaaa
     2881 agcattggtt cactttctgt aacacgacgt taaaccttcg tctccaatag gagcgctact
     2941 gattcaacat gccaatatat actaaatacg tttctacagt caaatgcttt aacgtttcat
     3001 gattaagtga ctatttaccg tcaatccttt cccattcctc ccactaatcc aactttttaa
     3061 ttactcttaa atcaccacta agcttcgaat ccatccaaaa ccacaatata aaaacagaac
     3121 tctcgtaact caatcatcgc aaaacaaaac aaaacaaaac aaaaacccca aaaagaaaga
     3181 ataagctaga tggattacaa ggaccacgac ggggattaca aggaccacga cattgattac
     3241 aaggatgatg atgacaagat ggctccgaag aagaagagga aggttggcat ccacggggtg
     3301 ccagctgctg acaagaagta ctcgatcggc ctcgatattg ggactaactc tgttggctgg
     3361 gccgtgatca ccgacgagta caaggtgccc tcaaagaagt tcaaggtcct gggcaacacc
     3421 gatcggcatt ccatcaagaa gaatctcatt ggcgctctcc tgttcgacag cggcgagacg
     3481 gctgaggcta cgcggctcaa gcgcaccgcc cgcaggcggt acacgcgcag gaagaatcgc
     3541 atctgctacc tgcaggagat tttctccaac gagatggcga aggttgacga ttctttcttc
     3601 cacaggctgg aggagtcatt cctcgtggag gaggataaga agcacgagcg gcatccaatc
     3661 ttcggcaaca ttgtcgacga ggttgcctac cacgagaagt accctacgat ctaccatctg
     3721 cggaagaagc tcgtggactc cacagataag gcggacctcc gcctgatcta cctcgctctg
     3781 gcccacatga ttaagttcag gggccatttc ctgatcgagg gggatctcaa cccggacaat
     3841 agcgatgttg acaagctgtt catccagctc gtgcagacgt acaaccagct cttcgaggag
     3901 aaccccatta atgcgtcagg cgtcgacgcg aaggctatcc tgtccgctag gctctcgaag
     3961 tctcggcgcc tcgagaacct gatcgcccag ctgccgggcg agaagaagaa cggcctgttc
     4021 gggaatctca ttgcgctcag cctggggctc acgcccaact tcaagtcgaa tttcgatctc
     4081 gctgaggacg ccaagctgca gctctccaag gacacatacg acgatgacct ggataacctc
     4141 ctggcccaga tcggcgatca gtacgcggac ctgttcctcg ctgccaagaa tctgtcggac
     4201 gccatcctcc tgtctgatat tctcagggtg aacaccgaga ttacgaaggc tccgctctca
     4261 gcctccatga tcaagcgcta cgacgagcac catcaggatc tgaccctcct gaaggcgctg
     4321 gtcaggcagc agctccccga gaagtacaag gagatcttct tcgatcagtc gaagaacggc
     4381 tacgctgggt acattgacgg cggggcctct caggaggagt tctacaagtt catcaagccg
     4441 attctggaga agatggacgg cacggaggag ctgctggtga agctcaatcg cgaggacctc
     4501 ctgaggaagc agcggacatt cgataacggc agcatcccac accagattca tctcggggag
     4561 ctgcacgcta tcctgaggag gcaggaggac ttctaccctt tcctcaagga taaccgcgag
     4621 aagatcgaga agattctgac tttcaggatc ccgtactacg tcggcccact cgctaggggc
     4681 aactcccgct tcgcttggat gacccgcaag tcagaggaga cgatcacgcc gtggaacttc
     4741 gaggaggtgg tcgacaaggg cgctagcgct cagtcgttca tcgagaggat gacgaatttc
     4801 gacaagaacc tgccaaatga gaaggtgctc cctaagcact cgctcctgta cgagtacttc
     4861 acagtctaca acgagctgac taaggtgaag tatgtgaccg agggcatgag gaagccggct
     4921 ttcctgtctg gggagcagaa gaaggccatc gtggacctcc tgttcaagac caaccggaag
     4981 gtcacggtta agcagctcaa ggaggactac ttcaagaaga ttgagtgctt cgattcggtc
     5041 gagatctctg gcgttgagga ccgcttcaac gcctccctgg ggacctacca cgatctcctg
     5101 aagatcatta aggataagga cttcctggac aacgaggaga atgaggatat cctcgaggac
     5161 attgtgctga cactcactct gttcgaggac cgggagatga tcgaggagcg cctgaagact
     5221 tacgcccatc tcttcgatga caaggtcatg aagcagctca agaggaggag gtacaccggc
     5281 tgggggaggc tgagcaggaa gctcatcaac ggcattcggg acaagcagtc cgggaagacg
     5341 atcctcgact tcctgaagag cgatggcttc gcgaaccgca atttcatgca gctgattcac
     5401 gatgacagcc tcacattcaa ggaggatatc cagaaggctc aggtgagcgg ccagggggac
     5461 tcgctgcacg agcatatcgc gaacctcgct ggctcgccag ctatcaagaa ggggattctg
     5521 cagaccgtga aggttgtgga cgagctggtg aaggtcatgg gcaggcacaa gcctgagaac
     5581 atcgtcattg agatggcccg ggagaatcag accacgcaga agggccagaa gaactcacgc
     5641 gagaggatga agaggatcga ggagggcatt aaggagctgg ggtcccagat cctcaaggag
     5701 cacccggtgg agaacacgca gctgcagaat gagaagctct acctgtacta cctccagaat
     5761 ggccgcgata tgtatgtgga ccaggagctg gatattaaca ggctcagcga ttacgacgtc
     5821 gatcatatcg ttccacagtc attcctgaag gatgactcca ttgacaacaa ggtcctcacc
     5881 aggtcggaca agaaccgggg caagtctgat aatgttcctt cagaggaggt cgttaagaag
     5941 atgaagaact actggcgcca gctcctgaat gccaagctga tcacgcagcg gaagttcgat
     6001 aacctcacaa aggctgagag gggcgggctc tctgagctgg acaaggcggg cttcatcaag
     6061 aggcagctgg tcgagacacg gcagatcact aagcacgttg cgcagattct cgactcacgg
     6121 atgaacacta agtacgatga gaatgacaag ctgatccgcg aggtgaaggt catcaccctg
     6181 aagtcaaagc tcgtctccga cttcaggaag gatttccagt tctacaaggt tcgggagatc
     6241 aacaattacc accatgccca tgacgcgtac ctgaacgcgg tggtcggcac agctctgatc
     6301 aagaagtacc caaagctcga gagcgagttc gtgtacgggg actacaaggt ttacgatgtg
     6361 aggaagatga tcgccaagtc ggagcaggag attggcaagg ctaccgccaa gtacttcttc
     6421 tactctaaca ttatgaattt cttcaagaca gagatcactc tggccaatgg cgagatccgg
     6481 aagcgccccc tcatcgagac gaacggcgag acgggggaga tcgtgtggga caagggcagg
     6541 gatttcgcga ccgtcaggaa ggttctctcc atgccacaag tgaatatcgt caagaagaca
     6601 gaggtccaga ctggcgggtt ctctaaggag tcaattctgc ctaagcggaa cagcgacaag
     6661 ctcatcgccc gcaagaagga ctgggatccg aagaagtacg gcgggttcga cagccccact
     6721 gtggcctact cggtcctggt tgtggcgaag gttgagaagg gcaagtccaa gaagctcaag
     6781 agcgtgaagg agctgctggg gatcacgatt atggagcgct ccagcttcga gaagaacccg
     6841 atcgatttcc tggaggcgaa gggctacaag gaggtgaaga aggacctgat cattaagctc
     6901 cccaagtact cactcttcga gctggagaac ggcaggaagc ggatgctggc ttccgctggc
     6961 gagctgcaga aggggaacga gctggctctg ccgtccaagt atgtgaactt cctctacctg
     7021 gcctcccact acgagaagct caagggcagc cccgaggaca acgagcagaa gcagctgttc
     7081 gtcgagcagc acaagcatta cctcgacgag atcattgagc agatttccga gttctccaag
     7141 cgcgtgatcc tggccgacgc gaatctggat aaggtcctct ccgcgtacaa caagcaccgc
     7201 gacaagccaa tcagggagca ggctgagaat atcattcatc tcttcaccct gacgaacctc
     7261 ggcgcccctg ctgctttcaa gtacttcgac acaactatcg atcgcaagag gtacacaagc
     7321 actaaggagg tcctggacgc gaccctcatc caccagtcga ttaccggcct ctacgagacg
     7381 cgcatcgacc tgtctcagct cgggggcgac aagcggccag cggcgacgaa gaaggcgggg
     7441 caggcgaaga agaagaagtg agctcagagc tttcgttcgt atcatcggtt tcgacaacgt
     7501 tcgtcaagtt caatgcatca gtttcattgc gcacacacca gaatcctact gagtttgagt
     7561 attatggcat tgggaaaact gtttttcttg taccatttgt tgtgcttgta atttactgtg
     7621 ttttttattc ggttttcgct atcgaactgt gaaatggaaa tggatggaga agagttaatg
     7681 aatgatatgg tccttttgtt cattctcaaa ttaatattat ttgttttttc tcttatttgt
     7741 tgtgtgttga atttgaaatt ataagagata tgcaaacatt ttgttttgag taaaaatgtg
     7801 tcaaatcgtg gcctctaatg accgaagtta atatgaggag taaaacactt gtagttgtac
     7861 cattatgctt attcactagg caacaaatat attttcagac ctagaaaagc tgcaaatgtt
     7921 actgaataca agtatgtcct cttgtgtttt agacatttat gaactttcct ttatgtaatt
     7981 ttccagaatc cttgtcagat tctaatcatt gctttataat tatagttata ctcatggatt
     8041 tgtagttgag tatgaaaata ttttttaatg cattttatga cttgccaatt gattgacaac
     8101 gaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     8161 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     8221 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     8281 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg ctagagcagc
     8341 ttgccaacat ggtggagcac gacactctcg tctactccaa gaatatcaaa gatacagtct
     8401 cagaagacca aagggctatt gagacttttc aacaaagggt aatatcggga aacctcctcg
     8461 gattccattg cccagctatc tgtcacttca tcaaaaggac agtagaaaag gaaggtggca
     8521 cctacaaatg ccatcattgc gataaaggaa aggctatcgt tcaagatgcc tctgccgaca
     8581 gtggtcccaa agatggaccc ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa
     8641 ccacgtcttc aaagcaagtg gattgatgtg aacatggtgg agcacgacac tctcgtctac
     8701 tccaagaata tcaaagatac agtctcagaa gaccaaaggg ctattgagac ttttcaacaa
     8761 agggtaatat cgggaaacct cctcggattc cattgcccag ctatctgtca cttcatcaaa
     8821 aggacagtag aaaaggaagg tggcacctac aaatgccatc attgcgataa aggaaaggct
     8881 atcgttcaag atgcctctgc cgacagtggt cccaaagatg gacccccacc cacgaggagc
     8941 atcgtggaaa aagaagacgt tccaaccacg tcttcaaagc aagtggattg atgtgatatc
     9001 tccactgacg taagggatga cgcacaatcc cactatcctt cgcaagaccc ttcctctata
     9061 taaggaagtt catttcattt ggagaggaca cgctgaaatc accagtctct ctctacaaat
     9121 ctatctctct cgagctttcg cagatctgtc gatcgaccat ggggattgaa caagatggat
     9181 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     9241 agacaatcgg ctgctctgat gccgccgtgt ttcggctgtc agcgcagggg cgcccggttc
     9301 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact ccaggacgag gcagcgcggc
     9361 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     9421 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     9481 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     9541 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     9601 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     9661 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     9721 cacatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     9781 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     9841 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     9901 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     9961 gactctgggg ttcggatcga tcctctagct agagtcgatc gacaagctcg agtttctcca
    10021 taataatgtg tgagtagttc ccagataagg gaattagggt tcctataggg tttcgctcat
    10081 gtgttgagca tataagaaac ccttagtatg tatttgtatt tgtaaaatac ttctatcaat
    10141 aaaatttcta attcctaaaa ccaaaatcca gtactaaaat ccagatcccc cgaattaatt
    10201 cggcgttaat tcagtacatt aaaaacgtcc gcaatgtgtt attaagttgt ctaagcgtca
    10261 atttgtttac accacaatat atcctgccac cagccagcca acagctcccc gaccggcagc
    10321 tcggcacaaa atcaccactc gatacaggca gcccatcagt ccgggacggc gtcagcggga
    10381 gagccgttgt aaggcggcag actttgctca tgttaccgat gctattcgga agaacggcaa
    10441 ctaagctgcc gggtttgaaa cacggatgat ctcgcggagg gtagcatgtt gattgtaacg
    10501 atgacagagc gttgctgcct gtgatcaccg cggtttcaaa atcggctccg tcgatactat
    10561 gttatacgcc aactttgaaa acaactttga aaaagctgtt ttctggtatt taaggtttta
    10621 gaatgcaagg aacagtgaat tggagttcgt cttgttataa ttagcttctt ggggtatctt
    10681 taaatactgt agaaaagagg aaggaaataa taaatggcta aaatgagaat atcaccggaa
    10741 ttgaaaaaac tgatcgaaaa ataccgctgc gtaaaagata cggaaggaat gtctcctgct
    10801 aaggtatata agctggtggg agaaaatgaa aacctatatt taaaaatgac ggacagccgg
    10861 tataaaggga ccacctatga tgtggaacgg gaaaaggaca tgatgctatg gctggaagga
    10921 aagctgcctg ttccaaaggt cctgcacttt gaacggcatg atggctggag caatctgctc
    10981 atgagtgagg ccgatggcgt cctttgctcg gaagagtatg aagatgaaca aagccctgaa
    11041 aagattatcg agctgtatgc ggagtgcatc aggctctttc actccatcga catatcggat
    11101 tgtccctata cgaatagctt agacagccgc ttagccgaat tggattactt actgaataac
    11161 gatctggccg atgtggattg cgaaaactgg gaagaagaca ctccatttaa agatccgcgc
    11221 gagctgtatg attttttaaa gacggaaaag cccgaagagg aacttgtctt ttcccacggc
    11281 gacctgggag acagcaacat ctttgtgaaa gatggcaaag taagtggctt tattgatctt
    11341 gggagaagcg gcagggcgga caagtggtat gacattgcct tctgcgtccg gtcgatcagg
    11401 gaggatatcg gggaagaaca gtatgtcgag ctattttttg acttactggg gatcaagcct
    11461 gattgggaga aaataaaata ttatatttta ctggatgaat tgttttagta cctagaatgc
    11521 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
    11581 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
    11641 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
    11701 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
    11761 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
    11821 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
    11881 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
    11941 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
    12001 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
    12061 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
    12121 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
    12181 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
    12241 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
    12301 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
    12361 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
    12421 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tacactccgc
    12481 tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
    12541 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
    12601 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ggtgccttga
    12661 tgtgggcgcc ggcggtcgag tggcgacggc gcggcttgtc cgcgccctgg tagattgcct
    12721 ggccgtaggc cagccatttt tgagcggcca gcggccgcga taggccgacg cgaagcggcg
    12781 gggcgtaggg agcgcagcga ccgaagggta ggcgcttttt gcagctcttc ggctgtgcgc
    12841 tggccagaca gttatgcaca ggccaggcgg gttttaagag ttttaataag ttttaaagag
    12901 ttttaggcgg aaaaatcgcc ttttttctct tttatatcag tcacttacat gtgtgaccgg
    12961 ttcccaatgt acggctttgg gttcccaatg tacgggttcc ggttcccaat gtacggcttt
    13021 gggttcccaa tgtacgtgct atccacagga aacagacctt ttcgaccttt ttcccctgct
    13081 agggcaattt gccctagcat ctgctccgta cattaggaac cggcggatgc ttcgccctcg
    13141 atcaggttgc ggtagcgcat gactaggatc gggccagcct gccccgcctc ctccttcaaa
    13201 tcgtactccg gcaggtcatt tgacccgatc agcttgcgca cggtgaaaca gaacttcttg
    13261 aactctccgg cgctgccact gcgttcgtag atcgtcttga acaaccatct ggcttctgcc
    13321 ttgcctgcgg cgcggcgtgc caggcggtag agaaaacggc cgatgccggg atcgatcaaa
    13381 aagtaatcgg ggtgaaccgt cagcacgtcc gggttcttgc cttctgtgat ctcgcggtac
    13441 atccaatcag ctagctcgat ctcgatgtac tccggccgcc cggtttcgct ctttacgatc
    13501 ttgtagcggc taatcaaggc ttcaccctcg gataccgtca ccaggcggcc gttcttggcc
    13561 ttcttcgtac gctgcatggc aacgtgcgtg gtgtttaacc gaatgcaggt ttctaccagg
    13621 tcgtctttct gctttccgcc atcggctcgc cggcagaact tgagtacgtc cgcaacgtgt
    13681 ggacggaaca cgcggccggg cttgtctccc ttcccttccc ggtatcggtt catggattcg
    13741 gttagatggg aaaccgccat cagtaccagg tcgtaatccc acacactggc catgccggcc
    13801 ggccctgcgg aaacctctac gtgcccgtct ggaagctcgt agcggatcac ctcgccagct
    13861 cgtcggtcac gcttcgacag acggaaaacg gccacgtcca tgatgctgcg actatcgcgg
    13921 gtgcccacgt catagagcat cggaacgaaa aaatctggtt gctcgtcgcc cttgggcggc
    13981 ttcctaatcg acggcgcacc ggctgccggc ggttgccggg attctttgcg gattcgatca
    14041 gcggccgctt gccacgattc accggggcgt gcttctgcct cgatgcgttg ccgctgggcg
    14101 gcctgcgcgg ccttcaactt ctccaccagg tcatcaccca gcgccgcgcc gatttgtacc
    14161 gggccggatg gtttgcgacc gctcacgccg attcctcggg cttgggggtt ccagtgccat
    14221 tgcagggccg gcaggcaacc cagccgctta cgcctggcca accgcccgtt cctccacaca
    14281 tggggcattc cacggcgtcg gtgcctggtt gttcttgatt ttccatgccg cctcctttag
    14341 ccgctaaaat tcatctactc atttattcat ttgctcattt actctggtag ctgcgcgatg
    14401 tattcagata gcagctcggt aatggtcttg ccttggcgta ccgcgtacat cttcagcttg
    14461 gtgtgatcct ccgccggcaa ctgaaagttg acccgcttca tggctggcgt gtctgccagg
    14521 ctggccaacg ttgcagcctt gctgctgcgt gcgctcggac ggccggcact tagcgtgttt
    14581 gtgcttttgc tcattttctc tttacctcat taactcaaat gagttttgat ttaatttcag
    14641 cggccagcgc ctggacctcg cgggcagcgt cgccctcggg ttctgattca agaacggttg
    14701 tgccggcggc ggcagtgcct gggtagctca cgcgctgcgt gatacgggac tcaagaatgg
    14761 gcagctcgta cccggccagc gcctcggcaa cctcaccgcc gatgcgcgtg cctttgatcg
    14821 cccgcgacac gacaaaggcc gcttgtagcc ttccatccgt gacctcaatg cgctgcttaa
    14881 ccagctccac caggtcggcg gtggcccata tgtcgtaagg gcttggctgc accggaatca
    14941 gcacgaagtc ggctgccttg atcgcggaca cagccaagtc cgccgcctgg ggcgctccgt
    15001 cgatcactac gaagtcgcgc cggccgatgg ccttcacgtc gcggtcaatc gtcgggcggt
    15061 cgatgccgac aacggttagc ggttgatctt cccgcacggc cgcccaatcg cgggcactgc
    15121 cctggggatc ggaatcgact aacagaacat cggccccggc gagttgcagg gcgcgggcta
    15181 gatgggttgc gatggtcgtc ttgcctgacc cgcctttctg gttaagtaca gcgataacct
    15241 tcatgcgttc cccttgcgta tttgtttatt tactcatcgc atcatatacg cagcgaccgc
    15301 atgacgcaag ctgttttact caaatacaca tcaccttttt agacggcggc gctcggtttc
    15361 ttcagcggcc aagctggccg gccaggccgc cagcttggca tcagacaaac cggccaggat
    15421 ttcatgcagc cgcacggttg agacgtgcgc gggcggctcg aacacgtacc cggccgcgat
    15481 catctccgcc tcgatctctt cggtaatgaa aaacggttcg tcctggccgt cctggtgcgg
    15541 tttcatgctt gttcctcttg gcgttcattc tcggcggccg ccagggcgtc ggcctcggtc
    15601 aatgcgtcct cacggaaggc accgcgccgc ctggcctcgg tgggcgtcac ttcctcgctg
    15661 cgctcaagtg cgcggtacag ggtcgagcga tgcacgccaa gcagtgcagc cgcctctttc
    15721 acggtgcggc cttcctggtc gatcagctcg cgggcgtgcg cgatctgtgc cggggtgagg
    15781 gtagggcggg ggccaaactt cacgcctcgg gccttggcgg cctcgcgccc gctccgggtg
    15841 cggtcgatga ttagggaacg ctcgaactcg gcaatgccgg cgaacacggt caacaccatg
    15901 cggccggccg gcgtggtggt gtcggcccac ggctctgcca ggctacgcag gcccgcgccg
    15961 gcctcctgga tgcgctcggc aatgtccagt aggtcgcggg tgctgcgggc caggcggtct
    16021 agcctggtca ctgtcacaac gtcgccaggg cgtaggtggt caagcatcct ggccagctcc
    16081 gggcggtcgc gcctggtgcc ggtgatcttc tcggaaaaca gcttggtgca gccggccgcg
    16141 tgcagttcgg cccgttggtt ggtcaagtcc tggtcgtcgg tgctgacgcg ggcatagccc
    16201 agcaggccag cggcggcgct cttgttcatg gcgtaatgtc tccggttcta gtcgcaagta
    16261 ttctacttta tgcgactaaa acacgcgaca agaaaacgcc aggaaaaggg cagggcggca
    16321 gcctgtcgcg taacttagga cttgtgcgac atgtcgtttt cagaagacgg ctgcactgaa
    16381 cgtcagaagc cgactgcact atagcagcgg aggggttgga tcaaagtact ttgatcccga
    16441 ggggaaccct gtggttggca tgcacataca aatggacgaa cggataaacc ttttcacgcc
    16501 cttttaaata tccgattatt ctaataaacg ctcttttctc ttag
//
