LOCUS       sequence-477292-       16690 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plant expression of Cas9 and gRNA against M. polymorpha CK2 beta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16690)
  TITLE     Hothorn CK2 plasmids
REFERENCE   2  (bases 1 to 16690)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: GE-0010-gRNA1-MpCKB
FEATURES             Location/Qualifiers
     source          1..16690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    54..78
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     266..288
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     280..297
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     281..297
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2057..2081
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2102..6205
                     /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"
     CDS             2102..6205
                     /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"
     CDS             6218..6238
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    complement(6741..6765)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    6794..6818
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        7369..7444
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(7486..7510)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     terminator      7550..7802
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(7835..7851)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7835..7851)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7848..7870)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7859..7875
                     /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(join(7883..7889,7890..7907,7908..7913))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7928..7949
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     polyA_signal    8034..8208
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(8249..9274)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     promoter        complement(9341..10017)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     complement(9384..9404)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    10212..10236
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     CDS             10757..11548
                     /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"
     rep_origin      11794..12382
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12283..12302
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12536..12553
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    12568..12708
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(12634..12653)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12794..12816
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      13052..13246
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(13312..14385)
                     /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(14814..15443)
                     /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 tttcacgccc ttttaaatat ccgattattc taataaacgc tcttttctct taggtttacc
       61 cgccaatata tcctgtcaaa cactgatagt ttaaactgaa ggcgggaaac gacaatctga
      121 tccaagctca agctgctcta gcattcgcca ttcaggctgc gcaactgttg ggaagggcga
      181 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
      241 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc
      301 aagcttggcc cgggcaaatg agtcacacac attgttgaga gacatatcaa aataactcaa
      361 ttgtaaggta agttgataga tttcccagtt aaatgcatta ttttaacaat atttgtatct
      421 caaattttca aaatcatatc aaatttgcag atcttgatga tgattcacaa ctctgtcttc
      481 gatttttgat tgttaacggg tatcgatatt tgggtaactt aaaaagaatt tgctaaagat
      541 cttaaaaaac agattaaaaa aacctttcaa aatagctaat cctttaaatc cgtttcacaa
      601 tataatgctc attgatgata atcgaaaaga ttccatttta aagtgcatat aaattgctta
      661 attatctctt tctgtctcgt aatattttgt tctctcctat tggatcggac ccgcatcttc
      721 tcgatcagtt cactatacta atatttggtc aacttcaaga gagattagaa cttcgttctc
      781 caaagtgtaa aggccgacaa gcagaaaagg aaacttcaat cagaagcctt ctaggttttc
      841 ccgcgtccca aaaagcgaag ggaggacggg gaggacggtg aggccagaag aagaaagcaa
      901 cagtaaaatc tcatgtgatt gcgtgcatgt catggatcta tgggatgcta gtccaccgtt
      961 tggaagtatg tgcagaagga gaaaggcgcg ggaggccaaa agcgacttcg gccacctctc
     1021 tactttctag aaagtttgct cgcagcttgt aataagcgcg gaaaaggtac tcgaaaaacc
     1081 ggagcaatcg gtgtggtatc agcagtcggg ttcgctaaac ctacgtcttc ttattttgga
     1141 atgttaaatt gaaagggcgt gccggcccta agcacggacg atgtgacaga cggaggagtg
     1201 tgagcaattc ggatgacgat gctggccggc agtcacgaca aagagtccga aggggcatgg
     1261 acagggcaat atataaatgg aaaagaggct ttgaatattc ctaacccttc gtcgtcggcc
     1321 tcccaacttg ccctcgcatg ctgaattggc cgcctatctg cgaatggccg gccgccgacc
     1381 ctaagtcgca ataacttgaa cggagccgat gcaaagtcct tatcgaacgc tcgttattat
     1441 aaacggggca gacctaggta tttaattata tcaaaaccct ccgcaatttc gctaatcatt
     1501 ccctaatttc ttctactttg gcgtctagtg ctgtggtcgc gcttgctctc aaagctcgct
     1561 gggaggcagt ggagcgtctg gcttaaaatc gatcgccgca accctagagt tcctttcact
     1621 tctttctctc ttgcgtggag agcatcgaca tcgaagagct gcatcttgtc ttctggtatg
     1681 cgccttgtgc cctcgtctct gtacgtcgtt acgattgctc gatattgtga ttgtctagac
     1741 ctgaaacccg actggagctt ttacgtggtg tcgattgggg cctcgagcga gtgatttttt
     1801 aggttcgttt tagccaagta catcccactt tggtcagtcc tgttatctcg gtaaggtgtt
     1861 cagttcggtc tctggtgcgg aggtgcctcg taagtttcct gcatgataga tttgtctatc
     1921 cttgacttga agaggagtcg aaggtcagcg aggttggctg tgtgctatgg acgtcggtgc
     1981 tccttggttc cgtgcttcta tgggtgttgc ctcaacagta ttgatgtgcc tgcagaaagg
     2041 ttgactagag ttatcaacaa gtttgtacaa aaaagcaggc tccgcggccg cccccttcac
     2101 catggataag aagtactcta tcggactcga tatcggaact aactctgtgg gatgggctgt
     2161 gatcaccgat gagtacaagg tgccatctaa gaagttcaag gttctcggaa acaccgatag
     2221 gcactctatc aagaaaaacc ttatcggtgc tctcctcttc gattctggtg aaactgctga
     2281 ggctaccaga ctcaagagaa ccgctagaag aaggtacacc agaagaaaga acaggatctg
     2341 ctacctccaa gagatcttct ctaacgagat ggctaaagtg gatgattcat tcttccacag
     2401 gctcgaagag tcattcctcg tggaagaaga taagaagcac gagaggcacc ctatcttcgg
     2461 aaacatcgtt gatgaggtgg cataccacga gaagtaccct actatctacc acctcagaaa
     2521 gaagctcgtt gattctactg ataaggctga tctcaggctc atctacctcg ctctcgctca
     2581 catgatcaag ttcagaggac acttcctcat cgagggtgat ctcaaccctg ataactctga
     2641 tgtggataag ttgttcatcc agctcgtgca gacctacaac cagcttttcg aagagaaccc
     2701 tatcaacgct tcaggtgtgg atgctaaggc tatcctctct gctaggctct ctaagtcaag
     2761 aaggcttgag aacctcattg ctcagctccc tggtgagaag aagaacggac ttttcggaaa
     2821 cttgatcgct ctctctctcg gactcacccc taacttcaag tctaacttcg atctcgctga
     2881 ggatgcaaag ctccagctct caaaggatac ctacgatgat gatctcgata acctcctcgc
     2941 tcagatcgga gatcagtacg ctgatttgtt cctcgctgct aagaacctct ctgatgctat
     3001 cctcctcagt gatatcctca gagtgaacac cgagatcacc aaggctccac tctcagcttc
     3061 tatgatcaag agatacgatg agcaccacca ggatctcaca cttctcaagg ctcttgttag
     3121 acagcagctc ccagagaagt acaaagagat tttcttcgat cagtctaaga acggatacgc
     3181 tggttacatc gatggtggtg catctcaaga agagttctac aagttcatca agcctatcct
     3241 cgagaagatg gatggaaccg aggaactcct cgtgaagctc aatagagagg atcttctcag
     3301 aaagcagagg accttcgata acggatctat ccctcatcag atccacctcg gagagttgca
     3361 cgctatcctt agaaggcaag aggatttcta cccattcctc aaggataaca gggaaaagat
     3421 tgagaagatt ctcaccttca gaatccctta ctacgtggga cctctcgcta gaggaaactc
     3481 aagattcgct tggatgacca gaaagtctga ggaaaccatc accccttgga acttcgaaga
     3541 ggtggtggat aagggtgcta gtgctcagtc tttcatcgag aggatgacca acttcgataa
     3601 gaaccttcca aacgagaagg tgctccctaa gcactctttg ctctacgagt acttcaccgt
     3661 gtacaacgag ttgaccaagg ttaagtacgt gaccgaggga atgaggaagc ctgctttttt
     3721 gtcaggtgag caaaagaagg ctatcgttga tctcttgttc aagaccaaca gaaaggtgac
     3781 cgtgaagcag ctcaaagagg attacttcaa gaaaatcgag tgcttcgatt cagttgagat
     3841 ttctggtgtt gaggataggt tcaacgcatc tctcggaacc taccacgatc tcctcaagat
     3901 cattaaggat aaggatttct tggataacga ggaaaacgag gatatcttgg aggatatcgt
     3961 tcttaccctc accctctttg aagatagaga gatgattgaa gaaaggctca agacctacgc
     4021 tcatctcttc gatgataagg tgatgaagca gttgaagaga agaagataca ctggttgggg
     4081 aaggctctca agaaagctca ttaacggaat cagggataag cagtctggaa agacaatcct
     4141 tgatttcctc aagtctgatg gattcgctaa cagaaacttc atgcagctca tccacgatga
     4201 ttctctcacc tttaaagagg atatccagaa ggctcaggtt tcaggacagg gtgatagtct
     4261 ccatgagcat atcgctaacc tcgctggatc tcctgcaatc aagaagggaa tcctccagac
     4321 tgtgaaggtt gtggatgagt tggtgaaggt gatgggaagg cataagcctg agaacatcgt
     4381 gatcgaaatg gctagagaga accagaccac tcagaaggga cagaagaact ctagggaaag
     4441 gatgaagagg atcgaggaag gtatcaaaga gcttggatct cagatcctca aagagcaccc
     4501 tgttgagaac actcagctcc agaatgagaa gctctacctc tactacctcc agaacggaag
     4561 ggatatgtat gtggatcaag agttggatat caacaggctc tctgattacg atgttgatca
     4621 tatcgtgcca cagtcattct tgaaggatga ttctatcgat aacaaggtgc tcaccaggtc
     4681 tgataagaac aggggtaaga gtgataacgt gccaagtgaa gaggttgtga agaaaatgaa
     4741 gaactattgg aggcagctcc tcaacgctaa gctcatcact cagagaaagt tcgataactt
     4801 gactaaggct gagaggggag gactctctga attggataag gcaggattca tcaagaggca
     4861 gcttgtggaa accaggcaga tcactaagca cgttgcacag atcctcgatt ctaggatgaa
     4921 caccaagtac gatgagaacg ataagttgat cagggaagtg aaggttatca ccctcaagtc
     4981 aaagctcgtg tctgatttca gaaaggattt ccaattctac aaggtgaggg aaatcaacaa
     5041 ctaccaccac gctcacgatg cttaccttaa cgctgttgtt ggaaccgctc tcatcaagaa
     5101 gtatcctaag ctcgagtcag agttcgtgta cggtgattac aaggtgtacg atgtgaggaa
     5161 gatgatcgct aagtctgagc aagagatcgg aaaggctacc gctaagtatt tcttctactc
     5221 taacatcatg aatttcttca agaccgagat taccctcgct aacggtgaga tcagaaagag
     5281 gccactcatc gagacaaacg gtgaaacagg tgagatcgtg tgggataagg gaagggattt
     5341 cgctaccgtt agaaaggtgc tctctatgcc acaggtgaac atcgttaaga aaaccgaggt
     5401 gcagaccggt ggattctcta aagagtctat cctccctaag aggaactctg ataagctcat
     5461 tgctaggaag aaggattggg accctaagaa atacggtggt ttcgattctc ctaccgtggc
     5521 ttactctgtt ctcgttgtgg ctaaggttga gaagggaaag agtaagaagc tcaagtctgt
     5581 taaggaactt ctcggaatca ctatcatgga aaggtcatct ttcgagaaga acccaatcga
     5641 tttcctcgag gctaagggat acaaagaggt taagaaggat ctcatcatca agctcccaaa
     5701 gtactcactc ttcgaactcg agaacggtag aaagaggatg ctcgcttctg ctggtgagct
     5761 tcaaaaggga aacgagcttg ctctcccatc taagtacgtt aactttcttt acctcgcttc
     5821 tcactacgag aagttgaagg gatctccaga agataacgag cagaagcaac ttttcgttga
     5881 gcagcacaag cactacttgg atgagatcat cgagcagatc tctgagttct ctaaaagggt
     5941 gatcctcgct gatgcaaacc tcgataaggt gttgtctgct tacaacaagc acagagataa
     6001 gcctatcagg gaacaggcag agaacatcat ccatctcttc acccttacca acctcggtgc
     6061 tcctgctgct ttcaagtact tcgatacaac catcgatagg aagagataca cctctaccaa
     6121 agaagtgctc gatgctaccc tcatccatca gtctatcact ggactctacg agactaggat
     6181 cgatctctca cagctcggtg gtgattcaag ggctgatcct aagaagaaga ggaaggtttg
     6241 aggcgcgccg agctccaggc ctcccagctt tcgtccgtat catcggtttc gacaacgttc
     6301 gtcaagttca atgcatcagt ttcattgccc acacaccaga atcctactaa gtttgagtat
     6361 tatggcattg gaaaagctgt tttcttctat catttgttct gcttgtaatt tactgtgttc
     6421 tttcagtttt tgttttcgga catcaaaatg caaatggatg gataagagtt aataaatgat
     6481 atggtccttt tgttcattct caaattatta ttatctgttg tttttacttt aatgggttga
     6541 atttaagtaa gaaaggaact aacagtgtga tattaaggtg caatgttaga catataaaac
     6601 agtctttcac ctctctttgg ttatgtcttg aattggtttg tttcttcact tatctgtgta
     6661 atcaagttta ctatgagtct atgatcaagt aattatgcaa tcaagttaag tacagtatag
     6721 gcttaagggt gggcgcgccg acccagcttt cttgtacaaa gtggttgata acagcggttg
     6781 actagagtta tcaacaagtt tgtacaaaaa agcaggctcc gcggccgccc ccttgtaacg
     6841 tgagactact acactcctca acgcaagttc tatctcatat ctctgcaaga gcttattcca
     6901 agaaactttc ttgtgcttct atgcatgccc aggtacctta agtcgcgcgc atagtcactt
     6961 cacagccttt tcagttctcc ctttgatggc tcagagtgac gcaactgcac aaaaataaga
     7021 aaaaagtcga agcgtcacga gagatgtatc cggctagttt ccttggagtt cggattgctc
     7081 tctttctctg tctccccgaa ccatatggaa tttatgaact gctgttcact agtcagtcac
     7141 ttgtttcaga acttttgagt ggtcgattta tccaccttct tgtttcttcc tgggtccaga
     7201 atctgacttc ttgagacaac aaacagtgca cagtggatcg aagaatgacc acagttgaga
     7261 cacgtggcgc tggcctcatt tattgcacca tacgtttagt gaagaactga ataatggtgg
     7321 atataaagca gagttgcacc cagcctctcg agaaaaggag aggccctcgt tttagagcta
     7381 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
     7441 gtgctttttt tctagtatcg ggaattctaa agggtgggcg cgccgaccca gctttcttgt
     7501 acaaagtggt tgataacagc gcttagaggc ttagagctcg aatttccccg atcgttcaaa
     7561 catttggcaa taaagtttct taagattgaa tcctgttgcc ggtcttgcga tgattatcat
     7621 ataatttctg ttgaattacg ttaagcatgt aataattaac atgtaatgca tgacgttatt
     7681 tatgagatgg gtttttatga ttagagtccc gcaattatac atttaatacg cgatagaaaa
     7741 caaaatatag cgcgcaaact aggataaatt atcgcgcgcg gtgtcatcta tgttactaga
     7801 tcgggaattg gttccggaac caattcgtaa tcatgtcata gctgtttcct gtgtgaaatt
     7861 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     7921 gtgcctaatg agtgagctaa ctcacattag gctgaattag gcgcgcctat ttctgaattc
     7981 aacacattgc ggacgttttt aatgtactga attaacgccg aattaattcg ggggatctgg
     8041 attttagtac tggattttgg ttttaggaat tagaaatttt attgatagaa gtattttaca
     8101 aatacaaata catactaagg gtttcttata tgctcaacac atgagcgaaa ccctatagga
     8161 accctaattc ccttatctgg gaactactca cacattatta tggagaaact cgagcttgtc
     8221 gatcgacaga tcccggtcgg catctactct atttctttgc cctcggacga gtgctggggc
     8281 gtcggtttcc actatcggcg agtacttcta cacagccatc ggtccagacg gccgcgcttc
     8341 tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt gcgtcgcatc
     8401 gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac caagctctga tagagttggt
     8461 caagaccaat gcggagcata tacgcccgga gtcgtggcga tcctgcaagc tccggatgcc
     8521 tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc cagaagaaga
     8581 tgttggcgac ctcgtattgg gaatccccga acatcgcctc gctccagtca atgaccgctg
     8641 ttatgcggcc attgtccgtc aggacattgt tggagccgaa atccgcgtgc acgaggtgcc
     8701 ggacttcggg gcagtcctcg gcccaaagca tcagctcatc gagagcctgc gcgacggacg
     8761 cactgacggt gtcgtccatc acagtttgcc agtgatacac atggggatca gcaatcgcgc
     8821 atatgaaatc acgccatgta gtgtattgac cgattccttg cggtccgaat gggccgaacc
     8881 cgctcgtctg gctaagatcg gccgcagcga tcgcatccat agcctccgcg accggttgta
     8941 gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt gaacggcggg
     9001 agatgcaata ggtcaggctc tcgctaaact ccccaatgtc aagcacttcc ggaatcggga
     9061 gcgcggccga tgcaaagtgc cgataaacat aacgatcttt gtagaaacca tcggcgcagc
     9121 tatttacccg caggacatat ccacgccctc ctacatcgaa gctgaaagca cgagattctt
     9181 cgccctccga gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc agaaacttct
     9241 cgacagacgt cgcggtgagt tcaggctttt tcatatctca ttgccccccg gatctgcgaa
     9301 agctcgagag agatagattt gtagagagag actggtgatt tcagcgtgtc ctctccaaat
     9361 gaaatgaact tccttatata gaggaagggt cttgcgaagg atagtgggat tgtgcgtcat
     9421 cccttacgtc agtggagata tcacatcaat ccacttgctt tgaagacgtg gttggaacgt
     9481 cttctttttc cacgatgctc ctcgtgggtg ggggtccatc tttgggacca ctgtcggcag
     9541 aggcatcttg aacgatagcc tttcctttat cgcaatgatg gcatttgtag gtgccacctt
     9601 ccttttctac tgtccttttg atgaagtgac agatagctgg gcaatggaat ccgaggaggt
     9661 ttcccgatat taccctttgt tgaaaagtct caatagccct ttggtcttct gagactgtat
     9721 ctttgatatt cttggagtag acgagagtgt cgtgctccac catgttcaca tcaatccact
     9781 tgctttgaag acgtggttgg aacgtcttct ttttccacga tgctcctcgt gggtgggggt
     9841 ccatctttgg gaccactgtc ggcagaggca tcttgaacga tagcctttcc tttatcgcaa
     9901 tgatggcatt tgtaggtgcc accttccttt tctactgtcc ttttgatgaa gtgacagata
     9961 gctgggcaat ggaatccgag gaggtttccc gatattaccc tttgttgaaa agtctcaata
    10021 gccctttggt cttctgagac tgtatctttg atattcttgg agtagacgag agtgtcgtgc
    10081 tccaccatgt tggcaagctg ctctagccaa tacgcaaacc gaattcagaa ataaattcag
    10141 cctaattcgg cgttaattca gtacattaaa aacgtccgca atgtgttatt aagttgtcta
    10201 agcgtcaatt tgtttacacc acaatatatc ctgccaccag ccagccaaca gctccccgac
    10261 cggcagctcg gcacaaaatc accactcgat acaggcagcc catcagtccg ggacggcgtc
    10321 agcgggagag ccgttgtaag gcggcagact ttgctcatgt taccgatgct attcggaaga
    10381 acggcaacta agctgccggg tttgaaacac ggatgatctc gcggagggta gcatgttgat
    10441 tgtaacgatg acagagcgtt gctgcctgtg atcaattcgg gcacgaaccc agtggacata
    10501 agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca actggtccag
    10561 aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga
    10621 ctgttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt
    10681 gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa
    10741 aacaaagtta aacatcatgg gggaagcggt gatcgccgaa gtatcgactc aactatcaga
    10801 ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg
    10861 ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg ttacggtgac
    10921 cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc
    10981 ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga
    11041 catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa
    11101 tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct
    11161 gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga
    11221 tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc
    11281 gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta
    11341 cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg
    11401 cctgccggcc cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga
    11461 agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgtccact acgtgaaagg
    11521 cgagatcacc aaggtagtcg gcaaataatg tctagctaga aattcgttca agccgacgcc
    11581 gcttcgcggc gcggcttaac tcaagcgtta gatgcactaa gcacataatt gctcacagcc
    11641 aaactatcag gtcaagtctg cttttattat ttttaagcgt gcataataag ccctacacaa
    11701 attgggagat atatcatgca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
    11761 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
    11821 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
    11881 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
    11941 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
    12001 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
    12061 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
    12121 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
    12181 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
    12241 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
    12301 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
    12361 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
    12421 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
    12481 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
    12541 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
    12601 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
    12661 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
    12721 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
    12781 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
    12841 gcgaggcagg gtgccttgat gtgggcgccg gcggtcgagt ggcgacggcg cggcttgtcc
    12901 gcgccctggt agattgcctg gccgtaggcc agccattttt gagcggccag cggccgcgat
    12961 aggccgacgc gaagcggcgg ggcgtaggga gcgcagcgac cgaagggtag gcgctttttg
    13021 cagctcttcg gctgtgcgct ggccagacag ttatgcacag gccaggcggg ttttaagagt
    13081 tttaataagt tttaaagagt tttaggcgga aaaatcgcct tttttctctt ttatatcagt
    13141 cacttacatg tgtgaccggt tcccaatgta cggctttggg ttcccaatgt acgggttccg
    13201 gttcccaatg tacggctttg ggttcccaat gtacgtgcta tccacaggaa agagaccttt
    13261 tcgacctttt tcccctgcta gggcaatttg ccctagcatc tgctccgtac attaggaacc
    13321 ggcggatgct tcgccctcga tcaggttgcg gtagcgcatg actaggatcg ggccagcctg
    13381 ccccgcctcc tccttcaaat cgtactccgg caggtcattt gacccgatca gcttgcgcac
    13441 ggtgaaacag aacttcttga actctccggc gctgccactg cgttcgtaga tcgtcttgaa
    13501 caaccatctg gcttctgcct tgcctgcggc gcggcgtgcc aggcggtaga gaaaacggcc
    13561 gatgccggga tcgatcaaaa agtaatcggg gtgaaccgtc agcacgtccg ggttcttgcc
    13621 ttctgtgatc tcgcggtaca tccaatcagc tagctcgatc tcgatgtact ccggccgccc
    13681 ggtttcgctc tttacgatct tgtagcggct aatcaaggct tcaccctcgg ataccgtcac
    13741 caggcggccg ttcttggcct tcttcgtacg ctgcatggca acgtgcgtgg tgtttaaccg
    13801 aatgcaggtt tctaccaggt cgtctttctg ctttccgcca tcggctcgcc ggcagaactt
    13861 gagtacgtcc gcaacgtgtg gacggaacac gcggccgggc ttgtctccct tcccttcccg
    13921 gtatcggttc atggattcgg ttagatggga aaccgccatc agtaccaggt cgtaatccca
    13981 cacactggcc atgccggccg gccctgcgga aacctctacg tgcccgtctg gaagctcgta
    14041 gcggatcacc tcgccagctc gtcggtcacg cttcgacaga cggaaaacgg ccacgtccat
    14101 gatgctgcga ctatcgcggg tgcccacgtc atagagcatc ggaacgaaaa aatctggttg
    14161 ctcgtcgccc ttgggcggct tcctaatcga cggcgcaccg gctgccggcg gttgccggga
    14221 ttctttgcgg attcgatcag cggccgcttg ccacgattca ccggggcgtg cttctgcctc
    14281 gatgcgttgc cgctgggcgg cctgcgcggc cttcaacttc tccaccaggt catcacccag
    14341 cgccgcgccg atttgtaccg ggccggatgg tttgcgaccg ctcacgccga ttcctcgggc
    14401 ttgggggttc cagtgccatt gcagggccgg cagacaaccc agccgcttac gcctggccaa
    14461 ccgcccgttc ctccacacat ggggcattcc acggcgtcgg tgcctggttg ttcttgattt
    14521 tccatgccgc ctcctttagc cgctaaaatt catctactca tttattcatt tgctcattta
    14581 ctctggtagc tgcgcgatgt attcagatag cagctcggta atggtcttgc cttggcgtac
    14641 cgcgtacatc ttcagcttgg tgtgatcctc cgccggcaac tgaaagttga cccgcttcat
    14701 ggctggcgtg tctgccaggc tggccaacgt tgcagccttg ctgctgcgtg cgctcggacg
    14761 gccggcactt agcgtgtttg tgcttttgct cattttctct ttacctcatt aactcaaatg
    14821 agttttgatt taatttcagc ggccagcgcc tggacctcgc gggcagcgtc gccctcgggt
    14881 tctgattcaa gaacggttgt gccggcggcg gcagtgcctg ggtagctcac gcgctgcgtg
    14941 atacgggact caagaatggg cagctcgtac ccggccagcg cctcggcaac ctcaccgccg
    15001 atgcgcgtgc ctttgatcgc ccgcgacacg acaaaggccg cttgtagcct tccatccgtg
    15061 acctcaatgc gctgcttaac cagctccacc aggtcggcgg tggcccatat gtcgtaaggg
    15121 cttggctgca ccggaatcag cacgaagtcg gctgccttga tcgcggacac agccaagtcc
    15181 gccgcctggg gcgctccgtc gatcactacg aagtcgcgcc ggccgatggc cttcacgtcg
    15241 cggtcaatcg tcgggcggtc gatgccgaca acggttagcg gttgatcttc ccgcacggcc
    15301 gcccaatcgc gggcactgcc ctggggatcg gaatcgacta acagaacatc ggccccggcg
    15361 agttgcaggg cgcgggctag atgggttgcg atggtcgtct tgcctgaccc gcctttctgg
    15421 ttaagtacag cgataacctt catgcgttcc ccttgcgtat ttgtttattt actcatcgca
    15481 tcatatacgc agcgaccgca tgacgcaagc tgttttactc aaatacacat caccttttta
    15541 gacggcggcg ctcggtttct tcagcggcca agctggccgg ccaggccgcc agcttggcat
    15601 cagacaaacc ggccaggatt tcatgcagcc gcacggttga gacgtgcgcg ggcggctcga
    15661 acacgtaccc ggccgcgatc atctccgcct cgatctcttc ggtaatgaaa aacggttcgt
    15721 cctggccgtc ctggtgcggt ttcatgcttg ttcctcttgg cgttcattct cggcggccgc
    15781 cagggcgtcg gcctcggtca atgcgtcctc acggaaggca ccgcgccgcc tggcctcggt
    15841 gggcgtcact tcctcgctgc gctcaagtgc gcggtacagg gtcgagcgat gcacgccaag
    15901 cagtgcagcc gcctctttca cggtgcggcc ttcctggtcg atcagctcgc gggcgtgcgc
    15961 gatctgtgcc ggggtgaggg tagggcgggg gccaaacttc acgcctcggg ccttggcggc
    16021 ctcgcgcccg ctccgggtgc ggtcgatgat tagggaacgc tcgaactcgg caatgccggc
    16081 gaacacggtc aacaccatgc ggccggccgg cgtggtggtg tcggcccacg gctctgccag
    16141 gctacgcagg cccgcgccgg cctcctggat gcgctcggca atgtccagta ggtcgcgggt
    16201 gctgcgggcc aggcggtcta gcctggtcac tgtcacaacg tcgccagggc gtaggtggtc
    16261 aagcatcctg gccagctccg ggcggtcgcg cctggtgccg gtgatcttct cggaaaacag
    16321 cttggtgcag ccggccgcgt gcagttcggc ccgttggttg gtcaagtcct ggtcgtcggt
    16381 gctgacgcgg gcatagccca gcaggccagc ggcggcgctc ttgttcatgg cgtaatgtct
    16441 ccggttctag tcgcaagtat tctactttat gcgactaaaa cacgcgacaa gaaaacgcca
    16501 ggaaaagggc agggcggcag cctgtcgcgt aacttaggac ttgtgcgaca tgtcgttttc
    16561 agaagacggc tgcactgaac gtcagaagcc gactgcacta tagcagcgga ggggttggat
    16621 caaagtactt tgatcccgag gggaaccctg tggttggcat gcacatacaa atggacgaac
    16681 ggataaacct
//
