LOCUS       sequence-105873-       10092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express sgRNA/PTG with rice snoRNA U3 promoter and Cas9 with rice 
            ubiquitin promoter for transient expression in protoplast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10092)
  AUTHORS   Xie K, Minkenberg B, Yang Y
  TITLE     Boosting CRISPR/Cas9 multiplex editing capability with the 
            endogenous tRNA-processing system.
  JOURNAL   Proc Natl Acad Sci U S A. 2015 Mar 17;112(11):3570-5. doi: 
            10.1073/pnas.1420294112. Epub 2015 Mar 2.
  PUBMED    25733849
REFERENCE   2  (bases 1 to 10092)
  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"; doi: 
            "10.1073/pnas.1420294112"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2015-03-17- 17"; volume: "112"; issue: "11"; pages: "3570-5"
COMMENT     Sequence Label: pRGE32
FEATURES             Location/Qualifiers
     source          1..10092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             26..49
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      71..323
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(332..349)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(332..348)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(341..363)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      561..1016
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(648..667)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     858..879
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1033..1052)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1152..1174
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1212..1230)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1298..1402
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1403..1471,1472..2263)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGTRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1621..1640)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2434..3022
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2923..2942
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3176..3193
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3310..3331
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3346..3351,3352..3369,3370..3376)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3384..3400
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     3389..3411
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3408..3424
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3408..3424
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3443..3823
                     /label=OsU3 promoter
                     /note="Oryza sativa (rice) snRNA U3 promoter"
     misc_RNA        3840..3915
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        3996..5705
                     /gene="RUBQ2"
                     /label=Ubi promoter
                     /note="promoter for the rice (Oryza sativa) polyubiquitin 
                     gene RUBQ2"
     intron          4744..5705
                     /note="intron upstream of the coding sequence for the rice 
                     polyubiquitin gene RUBQ2"
     protein_bind    5716..5740
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             5785..5850
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             5857..5877
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5902..10002
                     /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"
     CDS             5902..10002
                     /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"
     CDS             10003..10050
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     protein_bind    complement(join(10083..10092,1..15))
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 cttgtacaaa gtggttgata acagcgacta caaggatgac gatgacaagg cttagagctc
       61 gaatttcccc gatcgttcaa acatttggca ataaagtttc ttaagattga atcctgttgc
      121 cggtcttgcg atgattatca tataatttct gttgaattac gttaagcatg taataattaa
      181 catgtaatgc atgacgttat ttatgagatg ggtttttatg attagagtcc cgcaattata
      241 catttaatac gcgatagaaa acaaaatata gcgcgcaaac taggataaat tatcgcgcgc
      301 ggtgtcatct atgttactag atcgggaatt cactggccgt cgttttacaa cgtcgtgact
      361 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
      421 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
      481 gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
      541 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
      601 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
      661 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
      721 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
      781 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
      841 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
      901 gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
      961 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     1021 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     1081 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     1141 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     1201 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     1261 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     1321 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     1381 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     1441 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     1501 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     1561 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     1621 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     1681 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     1741 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     1801 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     1861 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     1921 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     1981 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     2041 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     2101 tggagccggt gagcgtggca ctcgcggtat cattgcagca ctggggccag atggtaagcc
     2161 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     2221 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     2281 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     2341 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     2401 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2461 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2521 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2581 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2641 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2701 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2761 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2821 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2881 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     2941 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3001 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3061 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3121 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3181 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     3241 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     3301 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     3361 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     3421 tgaccatgat tacgccaagc ttaaggaatc tttaaacata cgaacagatc acttaaagtt
     3481 cttctgaagc aacttaaagt tatcaggcat gcatggatct tggaggaatc agatgtgcag
     3541 tcagggacca tagcacaaga caggcgtctt ctactggtgc taccagcaaa tgctggaagc
     3601 cgggaacact gggtacgttg gaaaccacgt gatgtgaaga agtaagataa actgtaggag
     3661 aaaagcattt cgtagtgggc catgaagcct ttcaggacat gtattgcagt atgggccggc
     3721 ccattacgca attggacgac aacaaagact agtattagta ccacctcggc tatccacata
     3781 gatcaaagct gatttaaaag agttgtgcag atgatccgtg gcaggagacc gaggtctcgg
     3841 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
     3901 gcaccgagtc ggtgcttttt tgttttagag ctagaaatag caagttaaaa taaggctagt
     3961 ccgtttttag cgcgtgcatg cctgcaggtc cacaaattcg ggtcaaggcg gaagccagcg
     4021 cgccacccca cgtcagcaaa tacggaggcg cggggttgac ggcgtcaccc ggtcctaacg
     4081 gcgaccaaca aaccagccag aagaaattac agtaaaaaaa aagtaaattg cactttgatc
     4141 caccttttat tacctaagtc tcaatttgga tcacccttaa acctatcttt tcaatttggg
     4201 ccgggttgtg gtttggacta ccatgaacaa cttttcgtca tgtctaactt ccctttcagc
     4261 aaacatatga accatatata gaggagatcg gccgtatact agagctgatg tgtttaaggt
     4321 cgttgattgc acgagaaaaa aaaatccaaa tcgcaacaat agcaaattta tctggttcaa
     4381 agtgaaaaga tatgtttaaa ggtagtccaa agtaaaactt atagataata aaatgtggtc
     4441 caaagcgtaa ttcactcaaa aaaaatcaac gagacgtgta ccaaacggag acaaacggca
     4501 tcttctcgaa atttcccaac cgctcgctcg cccgcctcgt cttcccggaa accgcggtgg
     4561 tttcagcgtg gcggattctc caagcagacg gagacgtcac ggcacgggac tcctcccacc
     4621 acccaaccgc cataaatacc agccccctca tctcctctcc tcgcatcagc tccacccccg
     4681 aaaaatttct ccccaatctc gcgaggctct cgtcgtcgaa tcgaatcctc tcgcgtcctc
     4741 aaggtacgct gcttctcctc tcctcgcttc gtttcgattc gatttcggac gggtgaggtt
     4801 gttttgttgc tagatccgat tggtggttag ggttgtcgat gtgattatcg tgagatgttt
     4861 aggggttgta gatctgatgg ttgtgatttg ggcacggttg gttcgatagg tggaatcgtg
     4921 gttaggtttt gggattggat gttggttctg atgattgggg ggaattttta cggttagatg
     4981 aattgttgga tgattcgatt ggggaaatcg gtgtagatct gttggggaat tgtggaacta
     5041 gtcatgcctg agtgattggt gcgatttgta gcgtgttcca tcttgtaggc cttgttgcga
     5101 gcatgttcag atctactgtt ccgctcttga ttgagttatt ggtgccatgg gttggtgcaa
     5161 acacaggctt taatatgtta tatctgtttt gtgtttgatg tagatctgta gggtagttct
     5221 tcttagacat ggttcaatta tgtagcttgt gcgtttcgat ttgatttcat atgttcacag
     5281 attagataat gatgaactct tttaattaat tgtcaatggt aaataggaag tcttgtcgct
     5341 atatctgtca taatgatctc atgttactat ctgccagtaa tttatgctaa gaactatatt
     5401 agaatatcat gttacaatct gtagtaatat catgttacaa tctgtagttc atctatataa
     5461 tctattgtgg taatttcttt ttactatctg tgtgaagatt attgccacta gttcattcta
     5521 cttatttctg aagttcagga tacgtgtgct gttactacct atctgaatac atgtgtgatg
     5581 tgcctgttac tatctttttg aatacatgta tgttctgttg gaatatgttt gctgtttgat
     5641 ccgttgttgt gtccttaatc ttgtgctagt tcttacccta tctgtttggt gattatttct
     5701 tgcagatagt tatcaacaag tttgtacaaa aaagcaggct tcgaaggaga tagaaccaat
     5761 tctctaagga aatacttaac catggactat aaggaccacg acggagacta caaggatcat
     5821 gatattgatt acaaagacga tgacgataag atggccccaa agaagaagcg gaaggtcggt
     5881 atccacggag tcccagcagc cgacaagaag tacagcatcg gcctggacat cggcaccaac
     5941 tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg
     6001 ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac
     6061 agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga
     6121 cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac
     6181 gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag
     6241 cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc
     6301 atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc
     6361 tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg
     6421 aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag
     6481 ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc
     6541 agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag
     6601 aatggcctgt tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc
     6661 aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac
     6721 ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag
     6781 aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag
     6841 gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg
     6901 ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag
     6961 agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag
     7021 ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac
     7081 agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc
     7141 cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag
     7201 gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct
     7261 ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc
     7321 ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg
     7381 atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg
     7441 tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg
     7501 agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag
     7561 accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc
     7621 ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac
     7681 cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac
     7741 attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa
     7801 cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg
     7861 agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag
     7921 tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg
     7981 cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc
     8041 ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag
     8101 aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac
     8161 aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag
     8221 aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag
     8281 atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac
     8341 tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc
     8401 gactacgatg tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac
     8461 aaggtgctga ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag
     8521 gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag
     8581 agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc
     8641 ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc
     8701 ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa
     8761 gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa
     8821 gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga
     8881 accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag
     8941 gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc
     9001 aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac
     9061 ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg
     9121 gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc
     9181 gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg
     9241 aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc
     9301 gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc
     9361 aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc
     9421 gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg
     9481 atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg
     9541 gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac
     9601 ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag
     9661 aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc
     9721 gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac
     9781 aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc
     9841 ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag
     9901 aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc
     9961 ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acaaaaggcc ggcggccacg
    10021 aaaaaggccg gccaggcaaa aaagaaaaag taagaattcg cggccgcact cgagatatct
    10081 agacccagct tt
//
