pHSE401

LOCUS       pHSE401               16653 bp ds-DNA     circular SYN 13-NOV-2018
DEFINITION  CRISPR/Cas based plant genome editing and gene regulation; expresses
            3×FLAG-NLS-zCas9-NLS, gRNA scaffold for insertion of target sequence
            (AtU6-26 promoter), Hyg resistance.
ACCESSION   .
VERSION     .
KEYWORDS    pHSE401
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16653)
  AUTHORS   Xing HL, Dong L, Wang ZP, Zhang HY, Han CY, Liu B, Wang XC, Chen QJ
  TITLE     A CRISPR/Cas9 toolkit for multiplex genome editing in plants.
  JOURNAL   BMC Plant Biol. 2014 Nov 29;14(1):327.
  PUBMED    25432517
REFERENCE   2  (bases 1 to 16653)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 13, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..16653
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    21..45
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     233..255
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     247..264
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     248..264
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             977..1768
                     /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        1920..1995
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        2609..2953
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             3106..3171
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3178..3198
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3223..7323
                     /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             7236..7244
                     /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             7324..7371
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(8028..8044)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8028..8044)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8041..8063)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8052..8068
                     /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(8076..8106)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8121..8142
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8333..9009
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     CDS             9076..10101
                     /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"
     polyA_signal    10142..10316
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    10394..10418
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             10843..11637
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      11724..12312
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12213..12232
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12466..12483
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    12498..12638
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(12564..12583)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12724..12746
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      12982..13176
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(13242..14315)
                     /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(14744..15373)
                     /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 taaacgctct tttctcttag gtttacccgc caatatatcc tgtcaaacac tgatagttta
       61 aactgaaggc gggaaacgac aatctgatcc aagctcaagc tgctctagca ttcgccattc
      121 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      181 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      241 cgacgttgta aaacgacggc cagtgccaag cttcgacttg ccttccgcac aatacatcat
      301 ttcttcttag ctttttttct tcttcttcgt tcatacagtt tttttttgtt tatcagctta
      361 cattttcttg aaccgtagct ttcgttttct tctttttaac tttccattcg gagtttttgt
      421 atcttgtttc atagtttgtc ccaggattag aatgattagg catcgaacct tcaagaattt
      481 gattgaataa aacatcttca ttcttaagat atgaagataa tcttcaaaag gcccctggga
      541 atctgaaaga agagaagcag gcccatttat atgggaaaga acaatagtat ttcttatata
      601 ggcccattta agttgaaaac aatcttcaaa agtcccacat cgcttagata agaaaacgaa
      661 gctgagttta tatacagcta gagtcgaagt agtgattggg agaccaaccc agtggacata
      721 agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca actggtccag
      781 aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga
      841 ctgttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt
      901 gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa
      961 aacaaagtta aacatcatgg gggaagcggt gatcgccgaa gtatcgactc aactatcaga
     1021 ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg
     1081 ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg ttacggtgac
     1141 cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc
     1201 ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga
     1261 catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa
     1321 tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct
     1381 gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga
     1441 tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc
     1501 gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta
     1561 cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg
     1621 cctgccggcc cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga
     1681 agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgtccact acgtgaaagg
     1741 cgagatcacc aaggtagtcg gcaaataatg tctagctaga aattcgttca agccgacgcc
     1801 gcttcgcggc gcggcttaac tcaagcgtta gatgcactaa gcacataatt gctcacagcc
     1861 aaactatcag gtcaagtctg cttttattat ttttaagcgt gcataataag ccggtctcgg
     1921 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
     1981 gcaccgagtc ggtgcttttt tttgcaaaat tttccagatc gatttcttct tcctctgttc
     2041 ttcggcgttc aatttctggg gttttctctt cgttttctgt aactgaaacc taaaatttga
     2101 cctaaaaaaa atctcaaata atatgattca gtggttttgt acttttcagt tagttgagtt
     2161 ttgcagttcc gatgagataa accaataagc ttgcatgcct gcaggtcaac atggtggagc
     2221 acgacacact tgtctactcc aaaaatatca aagatacagt ctcagaagac caaagggcaa
     2281 ttgagacttt tcaacaaagg gtaatatccg gaaacctcct cggattccat tgcccagcta
     2341 tctgtcactt tattgtgaag atagtggaaa aggaaggtgg ctcctacaaa tgccatcatt
     2401 gcgataaagg aaaggccatc gttgaagatg cctctgccga cagtggtccc aaagatggac
     2461 ccccacccac gaggagcatc gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag
     2521 tggattgatg tgataacatg gtggagcacg acacacttgt ctactccaaa aatatcaaag
     2581 atacagtctc agaagaccaa agggcaattg agacttttca acaaagggta atatccggaa
     2641 acctcctcgg attccattgc ccagctatct gtcactttat tgtgaagata gtggaaaagg
     2701 aaggtggctc ctacaaatgc catcattgcg ataaaggaaa ggccatcgtt gaagatgcct
     2761 ctgccgacag tggtcccaaa gatggacccc cacccacgag gagcatcgtg gaaaaagaag
     2821 acgttccaac cacgtcttca aagcaagtgg attgatgtga tatctccact gacgtaaggg
     2881 atgacgcaca atcccactat ccttcgcaag acccttcctc tatataagga agttcatttc
     2941 atttggagag gacctcgacc tcaacacaac atatacaaaa caaacgaatc tcaagcaatc
     3001 aagcattcta cttctattgc agcaatttaa atcatttctt ttaaagcaaa agcaattttc
     3061 tgaaaatttt caccatttac gaacgatact cgagtaatct agatggatta caaggaccac
     3121 gacggggatt acaaggacca cgacattgat tacaaggatg atgatgacaa gatggctccg
     3181 aagaagaaga ggaaggttgg catccacggg gtgccagctg ctgacaagaa gtactcgatc
     3241 ggcctcgata ttgggactaa ctctgttggc tgggccgtga tcaccgacga gtacaaggtg
     3301 ccctcaaaga agttcaaggt cctgggcaac accgatcggc attccatcaa gaagaatctc
     3361 attggcgctc tcctgttcga cagcggcgag acggctgagg ctacgcggct caagcgcacc
     3421 gcccgcaggc ggtacacgcg caggaagaat cgcatctgct acctgcagga gattttctcc
     3481 aacgagatgg cgaaggttga cgattctttc ttccacaggc tggaggagtc attcctcgtg
     3541 gaggaggata agaagcacga gcggcatcca atcttcggca acattgtcga cgaggttgcc
     3601 taccacgaga agtaccctac gatctaccat ctgcggaaga agctcgtgga ctccacagat
     3661 aaggcggacc tccgcctgat ctacctcgct ctggcccaca tgattaagtt caggggccat
     3721 ttcctgatcg agggggatct caacccggac aatagcgatg ttgacaagct gttcatccag
     3781 ctcgtgcaga cgtacaacca gctcttcgag gagaacccca ttaatgcgtc aggcgtcgac
     3841 gcgaaggcta tcctgtccgc taggctctcg aagtctcggc gcctcgagaa cctgatcgcc
     3901 cagctgccgg gcgagaagaa gaacggcctg ttcgggaatc tcattgcgct cagcctgggg
     3961 ctcacgccca acttcaagtc gaatttcgat ctcgctgagg acgccaagct gcagctctcc
     4021 aaggacacat acgacgatga cctggataac ctcctggccc agatcggcga tcagtacgcg
     4081 gacctgttcc tcgctgccaa gaatctgtcg gacgccatcc tcctgtctga tattctcagg
     4141 gtgaacaccg agattacgaa ggctccgctc tcagcctcca tgatcaagcg ctacgacgag
     4201 caccatcagg atctgaccct cctgaaggcg ctggtcaggc agcagctccc cgagaagtac
     4261 aaggagatct tcttcgatca gtcgaagaac ggctacgctg ggtacattga cggcggggcc
     4321 tctcaggagg agttctacaa gttcatcaag ccgattctgg agaagatgga cggcacggag
     4381 gagctgctgg tgaagctcaa tcgcgaggac ctcctgagga agcagcggac attcgataac
     4441 ggcagcatcc cacaccagat tcatctcggg gagctgcacg ctatcctgag gaggcaggag
     4501 gacttctacc ctttcctcaa ggataaccgc gagaagatcg agaagattct gactttcagg
     4561 atcccgtact acgtcggccc actcgctagg ggcaactccc gcttcgcttg gatgacccgc
     4621 aagtcagagg agacgatcac gccgtggaac ttcgaggagg tggtcgacaa gggcgctagc
     4681 gctcagtcgt tcatcgagag gatgacgaat ttcgacaaga acctgccaaa tgagaaggtg
     4741 ctccctaagc actcgctcct gtacgagtac ttcacagtct acaacgagct gactaaggtg
     4801 aagtatgtga ccgagggcat gaggaagccg gctttcctgt ctggggagca gaagaaggcc
     4861 atcgtggacc tcctgttcaa gaccaaccgg aaggtcacgg ttaagcagct caaggaggac
     4921 tacttcaaga agattgagtg cttcgattcg gtcgagatct ctggcgttga ggaccgcttc
     4981 aacgcctccc tggggaccta ccacgatctc ctgaagatca ttaaggataa ggacttcctg
     5041 gacaacgagg agaatgagga tatcctcgag gacattgtgc tgacactcac tctgttcgag
     5101 gaccgggaga tgatcgagga gcgcctgaag acttacgccc atctcttcga tgacaaggtc
     5161 atgaagcagc tcaagaggag gaggtacacc ggctggggga ggctgagcag gaagctcatc
     5221 aacggcattc gggacaagca gtccgggaag acgatcctcg acttcctgaa gagcgatggc
     5281 ttcgcgaacc gcaatttcat gcagctgatt cacgatgaca gcctcacatt caaggaggat
     5341 atccagaagg ctcaggtgag cggccagggg gactcgctgc acgagcatat cgcgaacctc
     5401 gctggctcgc cagctatcaa gaaggggatt ctgcagaccg tgaaggttgt ggacgagctg
     5461 gtgaaggtca tgggcaggca caagcctgag aacatcgtca ttgagatggc ccgggagaat
     5521 cagaccacgc agaagggcca gaagaactca cgcgagagga tgaagaggat cgaggagggc
     5581 attaaggagc tggggtccca gatcctcaag gagcacccgg tggagaacac gcagctgcag
     5641 aatgagaagc tctacctgta ctacctccag aatggccgcg atatgtatgt ggaccaggag
     5701 ctggatatta acaggctcag cgattacgac gtcgatcata tcgttccaca gtcattcctg
     5761 aaggatgact ccattgacaa caaggtcctc accaggtcgg acaagaaccg gggcaagtct
     5821 gataatgttc cttcagagga ggtcgttaag aagatgaaga actactggcg ccagctcctg
     5881 aatgccaagc tgatcacgca gcggaagttc gataacctca caaaggctga gaggggcggg
     5941 ctctctgagc tggacaaggc gggcttcatc aagaggcagc tggtcgagac acggcagatc
     6001 actaagcacg ttgcgcagat tctcgactca cggatgaaca ctaagtacga tgagaatgac
     6061 aagctgatcc gcgaggtgaa ggtcatcacc ctgaagtcaa agctcgtctc cgacttcagg
     6121 aaggatttcc agttctacaa ggttcgggag atcaacaatt accaccatgc ccatgacgcg
     6181 tacctgaacg cggtggtcgg cacagctctg atcaagaagt acccaaagct cgagagcgag
     6241 ttcgtgtacg gggactacaa ggtttacgat gtgaggaaga tgatcgccaa gtcggagcag
     6301 gagattggca aggctaccgc caagtacttc ttctactcta acattatgaa tttcttcaag
     6361 acagagatca ctctggccaa tggcgagatc cggaagcgcc ccctcatcga gacgaacggc
     6421 gagacggggg agatcgtgtg ggacaagggc agggatttcg cgaccgtcag gaaggttctc
     6481 tccatgccac aagtgaatat cgtcaagaag acagaggtcc agactggcgg gttctctaag
     6541 gagtcaattc tgcctaagcg gaacagcgac aagctcatcg cccgcaagaa ggactgggat
     6601 ccgaagaagt acggcgggtt cgacagcccc actgtggcct actcggtcct ggttgtggcg
     6661 aaggttgaga agggcaagtc caagaagctc aagagcgtga aggagctgct ggggatcacg
     6721 attatggagc gctccagctt cgagaagaac ccgatcgatt tcctggaggc gaagggctac
     6781 aaggaggtga agaaggacct gatcattaag ctccccaagt actcactctt cgagctggag
     6841 aacggcagga agcggatgct ggcttccgct ggcgagctgc agaaggggaa cgagctggct
     6901 ctgccgtcca agtatgtgaa cttcctctac ctggcctccc actacgagaa gctcaagggc
     6961 agccccgagg acaacgagca gaagcagctg ttcgtcgagc agcacaagca ttacctcgac
     7021 gagatcattg agcagatttc cgagttctcc aagcgcgtga tcctggccga cgcgaatctg
     7081 gataaggtcc tctccgcgta caacaagcac cgcgacaagc caatcaggga gcaggctgag
     7141 aatatcattc atctcttcac cctgacgaac ctcggcgccc ctgctgcttt caagtacttc
     7201 gacacaacta tcgatcgcaa gaggtacaca agcactaagg aggtcctgga cgcgaccctc
     7261 atccaccagt cgattaccgg cctctacgag acgcgcatcg acctgtctca gctcgggggc
     7321 gacaagcggc cagcggcgac gaagaaggcg gggcaggcga agaagaagaa gtgagctcag
     7381 agctttcgtt cgtatcatcg gtttcgacaa cgttcgtcaa gttcaatgca tcagtttcat
     7441 tgcgcacaca ccagaatcct actgagtttg agtattatgg cattgggaaa actgtttttc
     7501 ttgtaccatt tgttgtgctt gtaatttact gtgtttttta ttcggttttc gctatcgaac
     7561 tgtgaaatgg aaatggatgg agaagagtta atgaatgata tggtcctttt gttcattctc
     7621 aaattaatat tatttgtttt ttctcttatt tgttgtgtgt tgaatttgaa attataagag
     7681 atatgcaaac attttgtttt gagtaaaaat gtgtcaaatc gtggcctcta atgaccgaag
     7741 ttaatatgag gagtaaaaca cttgtagttg taccattatg cttattcact aggcaacaaa
     7801 tatattttca gacctagaaa agctgcaaat gttactgaat acaagtatgt cctcttgtgt
     7861 tttagacatt tatgaacttt cctttatgta attttccaga atccttgtca gattctaatc
     7921 attgctttat aattatagtt atactcatgg atttgtagtt gagtatgaaa atatttttta
     7981 atgcatttta tgacttgcca attgattgac aacgaattcg taatcatgtc atagctgttt
     8041 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     8101 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     8161 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     8221 gggagaggcg gtttgcgtat tggctagagc agcttgccaa catggtggag cacgacactc
     8281 tcgtctactc caagaatatc aaagatacag tctcagaaga ccaaagggct attgagactt
     8341 ttcaacaaag ggtaatatcg ggaaacctcc tcggattcca ttgcccagct atctgtcact
     8401 tcatcaaaag gacagtagaa aaggaaggtg gcacctacaa atgccatcat tgcgataaag
     8461 gaaaggctat cgttcaagat gcctctgccg acagtggtcc caaagatgga cccccaccca
     8521 cgaggagcat cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa gtggattgat
     8581 gtgaacatgg tggagcacga cactctcgtc tactccaaga atatcaaaga tacagtctca
     8641 gaagaccaaa gggctattga gacttttcaa caaagggtaa tatcgggaaa cctcctcgga
     8701 ttccattgcc cagctatctg tcacttcatc aaaaggacag tagaaaagga aggtggcacc
     8761 tacaaatgcc atcattgcga taaaggaaag gctatcgttc aagatgcctc tgccgacagt
     8821 ggtcccaaag atggaccccc acccacgagg agcatcgtgg aaaaagaaga cgttccaacc
     8881 acgtcttcaa agcaagtgga ttgatgtgat atctccactg acgtaaggga tgacgcacaa
     8941 tcccactatc cttcgcaaga cccttcctct atataaggaa gttcatttca tttggagagg
     9001 acacgctgaa atcaccagtc tctctctaca aatctatctc tctcgagctt tcgcagatcc
     9061 ggggggcaat gagatatgaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg
     9121 atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt
     9181 gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat
     9241 ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg
     9301 gaagtgcttg acattgggga gtttagcgag agcctgacct attgcatctc ccgccgttca
     9361 cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct acaaccggtc
     9421 gcggaggcta tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca
     9481 ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct
     9541 gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg
     9601 caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg
     9661 cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt
     9721 gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg
     9781 aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag
     9841 cttgcaggat cgccacgact ccgggcgtat atgctccgca ttggtcttga ccaactctat
     9901 cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca
     9961 atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc
    10021 gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact
    10081 cgtccgaggg caaagaaata gagtagatgc cgaccgggat ctgtcgatcg acaagctcga
    10141 gtttctccat aataatgtgt gagtagttcc cagataaggg aattagggtt cctatagggt
    10201 ttcgctcatg tgttgagcat ataagaaacc cttagtatgt atttgtattt gtaaaatact
    10261 tctatcaata aaatttctaa ttcctaaaac caaaatccag tactaaaatc cagatccccc
    10321 gaattaattc ggcgttaatt cagtacatta aaaacgtccg caatgtgtta ttaagttgtc
    10381 taagcgtcaa tttgtttaca ccacaatata tcctgccacc agccagccaa cagctccccg
    10441 accggcagct cggcacaaaa tcaccactcg atacaggcag cccatcagtc cgggacggcg
    10501 tcagcgggag agccgttgta aggcggcaga ctttgctcat gttaccgatg ctattcggaa
    10561 gaacggcaac taagctgccg ggtttgaaac acggatgatc tcgcggaggg tagcatgttg
    10621 attgtaacga tgacagagcg ttgctgcctg tgatcaccgc ggtttcaaaa tcggctccgt
    10681 cgatactatg ttatacgcca actttgaaaa caactttgaa aaagctgttt tctggtattt
    10741 aaggttttag aatgcaagga acagtgaatt ggagttcgtc ttgttataat tagcttcttg
    10801 gggtatcttt aaatactgta gaaaagagga aggaaataat aaatggctaa aatgagaata
    10861 tcaccggaat tgaaaaaact gatcgaaaaa taccgctgcg taaaagatac ggaaggaatg
    10921 tctcctgcta aggtatataa gctggtggga gaaaatgaaa acctatattt aaaaatgacg
    10981 gacagccggt ataaagggac cacctatgat gtggaacggg aaaaggacat gatgctatgg
    11041 ctggaaggaa agctgcctgt tccaaaggtc ctgcactttg aacggcatga tggctggagc
    11101 aatctgctca tgagtgaggc cgatggcgtc ctttgctcgg aagagtatga agatgaacaa
    11161 agccctgaaa agattatcga gctgtatgcg gagtgcatca ggctctttca ctccatcgac
    11221 atatcggatt gtccctatac gaatagctta gacagccgct tagccgaatt ggattactta
    11281 ctgaataacg atctggccga tgtggattgc gaaaactggg aagaagacac tccatttaaa
    11341 gatccgcgcg agctgtatga ttttttaaag acggaaaagc ccgaagagga acttgtcttt
    11401 tcccacggcg acctgggaga cagcaacatc tttgtgaaag atggcaaagt aagtggcttt
    11461 attgatcttg ggagaagcgg cagggcggac aagtggtatg acattgcctt ctgcgtccgg
    11521 tcgatcaggg aggatatcgg ggaagaacag tatgtcgagc tattttttga cttactgggg
    11581 atcaagcctg attgggagaa aataaaatat tatattttac tggatgaatt gttttagtac
    11641 ctagaatgca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
    11701 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
    11761 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
    11821 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
    11881 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
    11941 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
    12001 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
    12061 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
    12121 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
    12181 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
    12241 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
    12301 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
    12361 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
    12421 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
    12481 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
    12541 caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
    12601 acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg
    12661 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
    12721 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagg
    12781 gtgccttgat gtgggcgccg gcggtcgagt ggcgacggcg cggcttgtcc gcgccctggt
    12841 agattgcctg gccgtaggcc agccattttt gagcggccag cggccgcgat aggccgacgc
    12901 gaagcggcgg ggcgtaggga gcgcagcgac cgaagggtag gcgctttttg cagctcttcg
    12961 gctgtgcgct ggccagacag ttatgcacag gccaggcggg ttttaagagt tttaataagt
    13021 tttaaagagt tttaggcgga aaaatcgcct tttttctctt ttatatcagt cacttacatg
    13081 tgtgaccggt tcccaatgta cggctttggg ttcccaatgt acgggttccg gttcccaatg
    13141 tacggctttg ggttcccaat gtacgtgcta tccacaggaa acagaccttt tcgacctttt
    13201 tcccctgcta gggcaatttg ccctagcatc tgctccgtac attaggaacc ggcggatgct
    13261 tcgccctcga tcaggttgcg gtagcgcatg actaggatcg ggccagcctg ccccgcctcc
    13321 tccttcaaat cgtactccgg caggtcattt gacccgatca gcttgcgcac ggtgaaacag
    13381 aacttcttga actctccggc gctgccactg cgttcgtaga tcgtcttgaa caaccatctg
    13441 gcttctgcct tgcctgcggc gcggcgtgcc aggcggtaga gaaaacggcc gatgccggga
    13501 tcgatcaaaa agtaatcggg gtgaaccgtc agcacgtccg ggttcttgcc ttctgtgatc
    13561 tcgcggtaca tccaatcagc tagctcgatc tcgatgtact ccggccgccc ggtttcgctc
    13621 tttacgatct tgtagcggct aatcaaggct tcaccctcgg ataccgtcac caggcggccg
    13681 ttcttggcct tcttcgtacg ctgcatggca acgtgcgtgg tgtttaaccg aatgcaggtt
    13741 tctaccaggt cgtctttctg ctttccgcca tcggctcgcc ggcagaactt gagtacgtcc
    13801 gcaacgtgtg gacggaacac gcggccgggc ttgtctccct tcccttcccg gtatcggttc
    13861 atggattcgg ttagatggga aaccgccatc agtaccaggt cgtaatccca cacactggcc
    13921 atgccggccg gccctgcgga aacctctacg tgcccgtctg gaagctcgta gcggatcacc
    13981 tcgccagctc gtcggtcacg cttcgacaga cggaaaacgg ccacgtccat gatgctgcga
    14041 ctatcgcggg tgcccacgtc atagagcatc ggaacgaaaa aatctggttg ctcgtcgccc
    14101 ttgggcggct tcctaatcga cggcgcaccg gctgccggcg gttgccggga ttctttgcgg
    14161 attcgatcag cggccgcttg ccacgattca ccggggcgtg cttctgcctc gatgcgttgc
    14221 cgctgggcgg cctgcgcggc cttcaacttc tccaccaggt catcacccag cgccgcgccg
    14281 atttgtaccg ggccggatgg tttgcgaccg ctcacgccga ttcctcgggc ttgggggttc
    14341 cagtgccatt gcagggccgg caggcaaccc agccgcttac gcctggccaa ccgcccgttc
    14401 ctccacacat ggggcattcc acggcgtcgg tgcctggttg ttcttgattt tccatgccgc
    14461 ctcctttagc cgctaaaatt catctactca tttattcatt tgctcattta ctctggtagc
    14521 tgcgcgatgt attcagatag cagctcggta atggtcttgc cttggcgtac cgcgtacatc
    14581 ttcagcttgg tgtgatcctc cgccggcaac tgaaagttga cccgcttcat ggctggcgtg
    14641 tctgccaggc tggccaacgt tgcagccttg ctgctgcgtg cgctcggacg gccggcactt
    14701 agcgtgtttg tgcttttgct cattttctct ttacctcatt aactcaaatg agttttgatt
    14761 taatttcagc ggccagcgcc tggacctcgc gggcagcgtc gccctcgggt tctgattcaa
    14821 gaacggttgt gccggcggcg gcagtgcctg ggtagctcac gcgctgcgtg atacgggact
    14881 caagaatggg cagctcgtac ccggccagcg cctcggcaac ctcaccgccg atgcgcgtgc
    14941 ctttgatcgc ccgcgacacg acaaaggccg cttgtagcct tccatccgtg acctcaatgc
    15001 gctgcttaac cagctccacc aggtcggcgg tggcccatat gtcgtaaggg cttggctgca
    15061 ccggaatcag cacgaagtcg gctgccttga tcgcggacac agccaagtcc gccgcctggg
    15121 gcgctccgtc gatcactacg aagtcgcgcc ggccgatggc cttcacgtcg cggtcaatcg
    15181 tcgggcggtc gatgccgaca acggttagcg gttgatcttc ccgcacggcc gcccaatcgc
    15241 gggcactgcc ctggggatcg gaatcgacta acagaacatc ggccccggcg agttgcaggg
    15301 cgcgggctag atgggttgcg atggtcgtct tgcctgaccc gcctttctgg ttaagtacag
    15361 cgataacctt catgcgttcc ccttgcgtat ttgtttattt actcatcgca tcatatacgc
    15421 agcgaccgca tgacgcaagc tgttttactc aaatacacat caccttttta gacggcggcg
    15481 ctcggtttct tcagcggcca agctggccgg ccaggccgcc agcttggcat cagacaaacc
    15541 ggccaggatt tcatgcagcc gcacggttga gacgtgcgcg ggcggctcga acacgtaccc
    15601 ggccgcgatc atctccgcct cgatctcttc ggtaatgaaa aacggttcgt cctggccgtc
    15661 ctggtgcggt ttcatgcttg ttcctcttgg cgttcattct cggcggccgc cagggcgtcg
    15721 gcctcggtca atgcgtcctc acggaaggca ccgcgccgcc tggcctcggt gggcgtcact
    15781 tcctcgctgc gctcaagtgc gcggtacagg gtcgagcgat gcacgccaag cagtgcagcc
    15841 gcctctttca cggtgcggcc ttcctggtcg atcagctcgc gggcgtgcgc gatctgtgcc
    15901 ggggtgaggg tagggcgggg gccaaacttc acgcctcggg ccttggcggc ctcgcgcccg
    15961 ctccgggtgc ggtcgatgat tagggaacgc tcgaactcgg caatgccggc gaacacggtc
    16021 aacaccatgc ggccggccgg cgtggtggtg tcggcccacg gctctgccag gctacgcagg
    16081 cccgcgccgg cctcctggat gcgctcggca atgtccagta ggtcgcgggt gctgcgggcc
    16141 aggcggtcta gcctggtcac tgtcacaacg tcgccagggc gtaggtggtc aagcatcctg
    16201 gccagctccg ggcggtcgcg cctggtgccg gtgatcttct cggaaaacag cttggtgcag
    16261 ccggccgcgt gcagttcggc ccgttggttg gtcaagtcct ggtcgtcggt gctgacgcgg
    16321 gcatagccca gcaggccagc ggcggcgctc ttgttcatgg cgtaatgtct ccggttctag
    16381 tcgcaagtat tctactttat gcgactaaaa cacgcgacaa gaaaacgcca ggaaaagggc
    16441 agggcggcag cctgtcgcgt aacttaggac ttgtgcgaca tgtcgttttc agaagacggc
    16501 tgcactgaac gtcagaagcc gactgcacta tagcagcgga ggggttggat caaagtactt
    16561 tgatcccgag gggaaccctg tggttggcat gcacatacaa atggacgaac ggataaacct
    16621 tttcacgccc ttttaaatat ccgattattc taa
//