Cas9 MDC32

LOCUS       Cas9 MDC32               14188 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  2x35S promoting Glycing Max codon optimized Cas9 with hygromycin 
            plant selection.
ACCESSION   .
VERSION     .
KEYWORDS    Cas9 MDC32
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14188)
  AUTHORS   Michno JM, Wang X, Liu J, Curtin SJ, Kono TJ, Stupar RM
  TITLE     CRISPR/Cas mutagenesis of soybean and Medicago truncatula using a 
            new web-tool and a modified Cas9 enzyme.
  JOURNAL   GM Crops Food. 2015;6(4):243-52. doi: 10.1080/21645698.2015.1106063.
  PUBMED    26479970
REFERENCE   2  (bases 1 to 14188)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..14188
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(11..27)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11..27)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(24..46)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    35..51
                     /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(59..89)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    104..125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        316..993
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             1061..2086
                     /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
                     K"
     polyA_signal    2126..2300
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2378..2402
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             2827..3621
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      3708..4296
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4197..4216
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4450..4467
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4482..4622
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4548..4567)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4708..4730
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      4966..5160
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(5226..6293)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     CDS             complement(6727..7356)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    8656..8680
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     8868..8890
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8882..8899
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8883..8899
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        9326..9670
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     9614..9634
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             9731..13834
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     RBS             13747..13755
                     /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             13841..13861
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
       61 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
      121 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
      181 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattggctag agcagcttgc
      241 caacatggtg gagcacgaca ctctcgtcta ctccaagaat atcaaagata cagtctcaga
      301 agaccaaagg gctattgaga cttttcaaca aagggtaata tcgggaaacc tcctcggatt
      361 ccattgccca gctatctgtc acttcatcaa aaggacagta gaaaaggaag gtggcaccta
      421 caaatgccat cattgcgata aaggaaaggc tatcgttcaa gatgcctctg ccgacagtgg
      481 tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg ttccaaccac
      541 gtcttcaaag caagtggatt gatgtgataa catggtggag cacgacactc tcgtctactc
      601 caagaatatc aaagatacag tctcagaaga ccaaagggct attgagactt ttcaacaaag
      661 ggtaatatcg ggaaacctcc tcggattcca ttgcccagct atctgtcact tcatcaaaag
      721 gacagtagaa aaggaaggtg gcacctacaa atgccatcat tgcgataaag gaaaggctat
      781 cgttcaagat gcctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat
      841 cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa gtggattgat gtgatatctc
      901 cactgacgta agggatgacg cacaatccca ctatccttcg caagaccttc ctctatataa
      961 ggaagttcat ttcatttgga gaggacacgc tgaaatcacc agtctctctc tacaaatcta
     1021 tctctctcga gctttcgcag atcccggggg gcaatgagat atgaaaaagc ctgaactcac
     1081 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
     1141 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
     1201 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
     1261 tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggagttta gcgagagcct
     1321 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga
     1381 actgcccgct gttctacaac cggtcgcgga ggctatggat gcgatcgctg cggccgatct
     1441 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
     1501 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
     1561 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
     1621 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga
     1681 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
     1741 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg
     1801 ctacttcgag cggaggcatc cggagcttgc aggatcgcca cgactccggg cgtatatgct
     1861 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
     1921 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
     1981 acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga
     2041 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag aaatagagta gatgccgacc
     2101 ggatctgtcg atcgacaagc tcgagtttct ccataataat gtgtgagtag ttcccagata
     2161 agggaattag ggttcctata gggtttcgct catgtgttga gcatataaga aacccttagt
     2221 atgtatttgt atttgtaaaa tacttctatc aataaaattt ctaattccta aaaccaaaat
     2281 ccagtactaa aatccagatc ccccgaatta attcggcgtt aattcagtac attaaaaacg
     2341 tccgcaatgt gttattaagt tgtctaagcg tcaatttgtt tacaccacaa tatatcctgc
     2401 caccagccag ccaacagctc cccgaccggc agctcggcac aaaatcacca ctcgatacag
     2461 gcagcccatc agtccgggac ggcgtcagcg ggagagccgt tgtaaggcgg cagactttgc
     2521 tcatgttacc gatgctattc ggaagaacgg caactaagct gccgggtttg aaacacggat
     2581 gatctcgcgg agggtagcat gttgattgta acgatgacag agcgttgctg cctgtgatca
     2641 ccgcggtttc aaaatcggct ccgtcgatac tatgttatac gccaactttg aaaacaactt
     2701 tgaaaaagct gttttctggt atttaaggtt ttagaatgca aggaacagtg aattggagtt
     2761 cgtcttgtta taattagctt cttggggtat ctttaaatac tgtagaaaag aggaaggaaa
     2821 taataaatgg ctaaaatgag aatatcaccg gaattgaaaa aactgatcga aaaataccgc
     2881 tgcgtaaaag atacggaagg aatgtctcct gctaaggtat ataagctggt gggagaaaat
     2941 gaaaacctat atttaaaaat gacggacagc cggtataaag ggaccaccta tgatgtggaa
     3001 cgggaaaagg acatgatgct atggctggaa ggaaagctgc ctgttccaaa ggtcctgcac
     3061 tttgaacggc atgatggctg gagcaatctg ctcatgagtg aggccgatgg cgtcctttgc
     3121 tcggaagagt atgaagatga acaaagccct gaaaagatta tcgagctgta tgcggagtgc
     3181 atcaggctct ttcactccat cgacatatcg gattgtccct atacgaatag cttagacagc
     3241 cgcttagccg aattggatta cttactgaat aacgatctgg ccgatgtgga ttgcgaaaac
     3301 tgggaagaag acactccatt taaagatccg cgcgagctgt atgatttttt aaagacggaa
     3361 aagcccgaag aggaacttgt cttttcccac ggcgacctgg gagacagcaa catctttgtg
     3421 aaagatggca aagtaagtgg ctttattgat cttgggagaa gcggcagggc ggacaagtgg
     3481 tatgacattg ccttctgcgt ccggtcgatc agggaggata tcggggaaga acagtatgtc
     3541 gagctatttt ttgacttact ggggatcaag cctgattggg agaaaataaa atattatatt
     3601 ttactggatg aattgtttta gtacctagaa tgcatgacca aaatccctta acgtgagttt
     3661 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3721 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3781 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3841 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3901 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3961 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4021 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     4081 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     4141 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4201 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4261 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4321 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4381 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4441 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     4501 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     4561 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     4621 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     4681 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     4741 catcaccgaa acgcgcgagg cagggtgcct tgatgtgggc gccggcggtc gagtggcgac
     4801 ggcgcggctt gtccgcgccc tggtagattg cctggccgta ggccagccat ttttgagcgg
     4861 ccagcggccg cgataggccg acgcgaagcg gcggggcgta gggagcgcag cgaccgaagg
     4921 gtaggcgctt tttgcagctc ttcggctgtg cgctggccag acagttatgc acaggccagg
     4981 cgggttttaa gagttttaat aagttttaaa gagttttagg cggaaaaatc gccttttttc
     5041 tcttttatat cagtcactta catgtgtgac cggttcccaa tgtacggctt tgggttccca
     5101 atgtacgggt tccggttccc aatgtacggc tttgggttcc caatgtacgt gctatccaca
     5161 ggaaagagac cttttcgacc tttttcccct gctagggcaa tttgccctag catctgctcc
     5221 gtacattagg aaccggcgga tgcttcgccc tcgatcaggt tgcggtagcg catgactagg
     5281 atcgggccag cctgccccgc ctcctccttc aaatcgtact ccggcaggtc atttgacccg
     5341 atcagcttgc gcacggtgaa acagaacttc ttgaactctc cggcgctgcc actgcgttcg
     5401 tagatcgtct tgaacaacca tctggcttct gccttgcctg cggcgcggcg tgccaggcgg
     5461 tagagaaaac ggccgatgcc gggatcgatc aaaaagtaat cggggtgaac cgtcagcacg
     5521 tccgggttct tgccttctgt gatctcgcgg tacatccaat cagctagctc gatctcgatg
     5581 tactccggcc gcccggtttc gctctttacg atcttgtagc ggctaatcaa ggcttcaccc
     5641 tcggataccg tcaccaggcg gccgttcttg gccttcttcg tacgctgcat ggcaacgtgc
     5701 gtggtgttta accgaatgca ggtttctacc aggtcgtctt tctgctttcc gccatcggct
     5761 cgccggcaga acttgagtac gtccgcaacg tgtggacgga acacgcggcc gggcttgtct
     5821 cccttccctt cccggtatcg gttcatggat tcggttagat gggaaaccgc catcagtacc
     5881 aggtcgtaat cccacacact ggccatgccg gccggccctg cggaaacctc tacgtgcccg
     5941 tctggaagct cgtagcggat cacctcgcca gctcgtcggt cacgcttcga cagacggaaa
     6001 acggccacgt ccatgatgct gcgactatcg cgggtgccca cgtcatagag catcggaacg
     6061 aaaaaatctg gttgctcgtc gcccttgggc ggcttcctaa tcgacggcgc accggctgcc
     6121 ggcggttgcc gggattcttt gcggattcga tcagcggccg cttgccacga ttcaccgggg
     6181 cgtgcttctg cctcgatgcg ttgccgctgg gcggcctgcg cggccttcaa cttctccacc
     6241 aggtcatcac ccagcgccgc gccgatttgt accgggccgg atggtttgcg accgtcacgc
     6301 cgattcctcg ggcttggggg ttccagtgcc attgcagggc cggcagacaa cccagccgct
     6361 tacgcctggc caaccgcccg ttcctccaca catggggcat tccacggcgt cggtgcctgg
     6421 ttgttcttga ttttccatgc cgcctccttt agccgctaaa attcatctac tcatttattc
     6481 atttgctcat ttactctggt agctgcgcga tgtattcaga tagcagctcg gtaatggtct
     6541 tgccttggcg taccgcgtac atcttcagct tggtgtgatc ctccgccggc aactgaaagt
     6601 tgacccgctt catggctggc gtgtctgcca ggctggccaa cgttgcagcc ttgctgctgc
     6661 gtgcgctcgg acggccggca cttagcgtgt ttgtgctttt gctcattttc tctttacctc
     6721 attaactcaa atgagttttg atttaatttc agcggccagc gcctggacct cgcgggcagc
     6781 gtcgccctcg ggttctgatt caagaacggt tgtgccggcg gcggcagtgc ctgggtagct
     6841 cacgcgctgc gtgatacggg actcaagaat gggcagctcg tacccggcca gcgcctcggc
     6901 aacctcaccg ccgatgcgcg tgcctttgat cgcccgcgac acgacaaagg ccgcttgtag
     6961 ccttccatcc gtgacctcaa tgcgctgctt aaccagctcc accaggtcgg cggtggccca
     7021 tatgtcgtaa gggcttggct gcaccggaat cagcacgaag tcggctgcct tgatcgcgga
     7081 cacagccaag tccgccgcct ggggcgctcc gtcgatcact acgaagtcgc gccggccgat
     7141 ggccttcacg tcgcggtcaa tcgtcgggcg gtcgatgccg acaacggtta gcggttgatc
     7201 ttcccgcacg gccgcccaat cgcgggcact gccctgggga tcggaatcga ctaacagaac
     7261 atcggccccg gcgagttgca gggcgcgggc tagatgggtt gcgatggtcg tcttgcctga
     7321 cccgcctttc tggttaagta cagcgataac cttcatgcgt tccccttgcg tatttgttta
     7381 tttactcatc gcatcatata cgcagcgacc gcatgacgca agctgtttta ctcaaataca
     7441 catcaccttt ttagacggcg gcgctcggtt tcttcagcgg ccaagctggc cggccaggcc
     7501 gccagcttgg catcagacaa accggccagg atttcatgca gccgcacggt tgagacgtgc
     7561 gcgggcggct cgaacacgta cccggccgcg atcatctccg cctcgatctc ttcggtaatg
     7621 aaaaacggtt cgtcctggcc gtcctggtgc ggtttcatgc ttgttcctct tggcgttcat
     7681 tctcggcggc cgccagggcg tcggcctcgg tcaatgcgtc ctcacggaag gcaccgcgcc
     7741 gcctggcctc ggtgggcgtc acttcctcgc tgcgctcaag tgcgcggtac agggtcgagc
     7801 gatgcacgcc aagcagtgca gccgcctctt tcacggtgcg gccttcctgg tcgatcagct
     7861 cgcgggcgtg cgcgatctgt gccggggtga gggtagggcg ggggccaaac ttcacgcctc
     7921 gggccttggc ggcctcgcgc ccgctccggg tgcggtcgat gattagggaa cgctcgaact
     7981 cggcaatgcc ggcgaacacg gtcaacacca tgcggccggc cggcgtggtg gtgtcggccc
     8041 acggctctgc caggctacgc aggcccgcgc cggcctcctg gatgcgctcg gcaatgtcca
     8101 gtaggtcgcg ggtgctgcgg gccaggcggt ctagcctggt cactgtcaca acgtcgccag
     8161 ggcgtaggtg gtcaagcatc ctggccagct ccgggcggtc gcgcctggtg ccggtgatct
     8221 tctcggaaaa cagcttggtg cagccggccg cgtgcagttc ggcccgttgg ttggtcaagt
     8281 cctggtcgtc ggtgctgacg cgggcatagc ccagcaggcc agcggcggcg ctcttgttca
     8341 tggcgtaatg tctccggttc tagtcgcaag tattctactt tatgcgacta aaacacgcga
     8401 caagaaaacg ccaggaaaag ggcagggcgg cagcctgtcg cgtaacttag gacttgtgcg
     8461 acatgtcgtt ttcagaagac ggctgcactg aacgtcagaa gccgactgca ctatagcagc
     8521 ggaggggttg gatcaaagta ctttgatccc gaggggaacc ctgtggttgg catgcacata
     8581 caaatggacg aacggataaa ccttttcacg cccttttaaa tatccgttat tctaataaac
     8641 gctcttttct cttaggttta cccgccaata tatcctgtca aacactgata gtttaaactg
     8701 aaggcgggaa acgacaatct gatccaagct caagctgctc tagcattcgc cattcaggct
     8761 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     8821 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     8881 ttgtaaaacg acggccagtg ccaagcttgg cgtgcctgca ggtcaacatg gtggagcacg
     8941 acacacttgt ctactccaaa aatatcaaag atacagtctc agaagaccaa agggcaattg
     9001 agacttttca acaaagggta atatccggaa acctcctcgg attccattgc ccagctatct
     9061 gtcactttat tgtgaagata gtggaaaagg aaggtggctc ctacaaatgc catcattgcg
     9121 ataaaggaaa ggccatcgtt gaagatgcct ctgccgacag tggtcccaaa gatggacccc
     9181 cacccacgag gagcatcgtg gaaaaagaag acgttccaac cacgtcttca aagcaagtgg
     9241 attgatgtga taacatggtg gagcacgaca cacttgtcta ctccaaaaat atcaaagata
     9301 cagtctcaga agaccaaagg gcaattgaga cttttcaaca aagggtaata tccggaaacc
     9361 tcctcggatt ccattgccca gctatctgtc actttattgt gaagatagtg gaaaaggaag
     9421 gtggctccta caaatgccat cattgcgata aaggaaaggc catcgttgaa gatgcctctg
     9481 ccgacagtgg tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg
     9541 ttccaaccac gtcttcaaag caagtggatt gatgtgatat ctccactgac gtaagggatg
     9601 acgcacaatc ccactatcct tcgcaagacc cttcctctat ataaggaagt tcatttcatt
     9661 tggagaggac ctcgactcta gaggatcccc gggtaccggg ccccccctcg aggcgcgcca
     9721 ccaggtcgac atggataaga agtactccat cggactggat atcggaacta actccgtggg
     9781 atgggctgtg atcactgatg agtacaaggt gccatccaag aagttcaagg tgctgggaaa
     9841 cactgataga cactccatca agaagaacct gatcggagct ctgctgttcg attccggaga
     9901 gactgctgag gctactagac tgaagagaac tgctagaaga agatacacta gaagaaagaa
     9961 cagaatctgc tacctgcaag agatcttctc caacgagatg gctaaggtgg atgattcctt
    10021 cttccacaga ctggaggagt ccttcctggt ggaggaggat aagaagcacg agagacaccc
    10081 aatcttcgga aacatcgtgg atgaggtggc ttaccacgag aagtacccaa ctatctacca
    10141 cctgagaaag aagctggtgg attccactga taaggctgat ctgagactga tctacctggc
    10201 tctggctcac atgatcaagt tcagaggaca cttcctgatc gagggagatc tgaacccaga
    10261 taactccgat gtggataagc tgttcatcca actggtgcaa acttacaacc aactgttcga
    10321 ggagaaccca atcaacgctt ccggagtgga tgctaaggct atcctgtccg ctagactgtc
    10381 caagtccaga agactggaga acctgatcgc tcaactgcca ggagagaaga agaacggact
    10441 gttcggaaac ctgatcgctc tgtccctggg actgactcca aacttcaagt ccaacttcga
    10501 tctggctgag gatgctaagc tgcaactgtc caaggatact tacgatgatg atctggataa
    10561 cctgctggct caaatcggag atcaatacgc tgatctgttc ctggctgcta agaacctgtc
    10621 cgatgctatc ctgctgtccg atatcctgag agtgaacact gagatcacta aggctccact
    10681 gtccgcttcc atgatcaaga gatacgatga gcaccaccaa gatctgactc tgctgaaggc
    10741 tctggtgaga caacaactgc cagagaagta caaggagatc ttcttcgatc aatccaagaa
    10801 cggatacgct ggatacatcg atggaggagc ttcccaagag gagttctaca agttcatcaa
    10861 gccaatcctg gagaagatgg atggaactga ggagctgctg gtgaagctga acagagagga
    10921 tctgctgaga aagcaaagaa ctttcgataa cggatccatc ccacaccaaa tccacctggg
    10981 agagctgcac gctatcctga gaagacaaga ggatttctac ccattcctga aggataacag
    11041 agagaagatc gagaagatcc tgactttcag aatcccatac tacgtgggac cactggctag
    11101 aggaaactcc agattcgctt ggatgactag aaagtccgag gagactatca ctccatggaa
    11161 cttcgaggag gtggtggata agggagcttc cgctcaatcc ttcatcgaga gaatgactaa
    11221 cttcgataag aacctgccaa acgagaaggt gctgccaaag cactccctgc tgtacgagta
    11281 cttcactgtg tacaacgagc tgactaaggt gaagtacgtg actgagggaa tgagaaagcc
    11341 agctttcctg tccggagagc aaaagaaggc tatcgtggat ctgctgttca agactaacag
    11401 aaaggtgact gtgaagcaac tgaaggagga ttacttcaag aagatcgagt gcttcgattc
    11461 cgtggagatc tccggagtgg aggatagatt caacgcttcc ctgggaactt accacgatct
    11521 gctgaagatc atcaaggata aggatttcct ggataacgag gagaacgagg atatcctgga
    11581 ggatatcgtg ctgactctga ctctgttcga ggatagagag atgatcgagg agagactgaa
    11641 gacttacgct cacctgttcg atgataaggt gatgaagcaa ctgaagagaa gaagatacac
    11701 tggatgggga agactgtcca gaaagctgat caacggaatc agagataagc aatccggaaa
    11761 gactatcctg gatttcctga agtccgatgg attcgctaac agaaacttca tgcaactgat
    11821 ccacgatgat tccctgactt tcaaggagga tatccaaaag gctcaagtgt ccggacaagg
    11881 agattccctg cacgagcaca tcgctaacct ggctggatcc ccagctatca agaagggaat
    11941 cctgcaaact gtgaaggtgg tggatgagct ggtgaaggtg atgggaagac acaagccaga
    12001 gaacatcgtg atcgagatgg ctagagagaa ccaaactact caaaagggac aaaagaactc
    12061 cagagagaga atgaagagaa tcgaggaggg aatcaaggag ctgggatccc aaatcctgaa
    12121 ggagcaccca gtggagaaca ctcaactgca aaacgagaag ctgtacctgt actacctgca
    12181 aaacggaaga gatatgtacg tggatcaaga gctggatatc aacagactgt ccgattacga
    12241 tgtggatcac atcgtgccac aatccttcct gaaggatgat tccatcgata acaaggtgct
    12301 gactagatcc gataagaaca gaggaaagtc cgataacgtg ccatccgagg aggtggtgaa
    12361 gaagatgaag aactactgga gacaactgct gaacgctaag ctgatcactc aaagaaagtt
    12421 cgataacctg actaaggctg agagaggagg actgtccgag ctggataagg ctggattcat
    12481 caagagacaa ctggtggaga ctagacaaat cactaagcac gtggctcaaa tcctggattc
    12541 cagaatgaac actaagtacg atgagaacga taagctgatc agagaggtga aggtgatcac
    12601 tctgaagtcc aagctggtgt ccgatttcag aaaggatttc caattctaca aggtgagaga
    12661 gatcaacaac taccaccacg ctcacgatgc ttacctgaac gctgtggtgg gaactgctct
    12721 gatcaagaag tacccaaagc tggagtccga gttcgtgtac ggagattaca aggtgtacga
    12781 tgtgagaaag atgatcgcta agtccgagca agagatcgga aaggctactg ctaagtactt
    12841 cttctactcc aacatcatga acttcttcaa gactgagatc actctggcta acggagagat
    12901 cagaaagaga ccactgatcg agactaacgg agagactgga gagatcgtgt gggataaggg
    12961 aagagatttc gctactgtga gaaaggtgct gtccatgcca caagtgaaca tcgtgaagaa
    13021 gactgaggtg caaactggag gattctccaa ggagtccatc ctgccaaaga gaaactccga
    13081 taagctgatc gctagaaaga aggattggga tccaaagaag tacggaggat tcgattcccc
    13141 aactgtggct tactccgtgc tggtggtggc taaggtggag aagggaaagt ccaagaagct
    13201 gaagtccgtg aaggagctgc tgggaatcac tatcatggag agatcctcct tcgagaagaa
    13261 cccaatcgat ttcctggagg ctaagggata caaggaggtg aagaaggatc tgatcatcaa
    13321 gctgccaaag tactccctgt tcgagctgga gaacggaaga aagagaatgc tggcttccgc
    13381 tggagagctg caaaagggaa acgagctggc tctgccatcc aagtacgtga acttcctgta
    13441 cctggcttcc cactacgaga agctgaaggg atccccagag gataacgagc aaaagcaact
    13501 gttcgtggag caacacaagc actacctgga tgagatcatc gagcaaatct ccgagttctc
    13561 caagagagtg atcctggctg atgctaacct ggataaggtg ctgtccgctt acaacaagca
    13621 cagagataag ccaatcagag agcaagctga gaacatcatc cacctgttca ctctgactaa
    13681 cctgggagct ccagctgctt tcaagtactt cgatactact atcgatagaa agagatacac
    13741 ttccactaag gaggtgctgg atgctactct gatccaccaa tccatcactg gactgtacga
    13801 gactagaatc gatctgtccc aactgggagg agatctgcaa ccaaagaaga agagaaaggt
    13861 gggaggatga tctagaccag attaattaac tagttctaga gcggccgccc accgcggtgg
    13921 agctcgaatt tccccgatcg ttcaaacatt tggcaataaa gtttcttaag attgaatcct
    13981 gttgccggtc ttgcgatgat tatcatataa tttctgttga attacgttaa gcatgtaata
    14041 attaacatgt aatgcatgac gttatttatg agatgggttt ttatgattag agtcccgcaa
    14101 ttatacattt aatacgcgat agaaaacaaa atatagcgcg caaactagga taaattatcg
    14161 cgcgcggtgt catctatgtt actgaatt
//