pBUN411

LOCUS       pBUN411               13518 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  CRISPR/Cas based plant genome editing and gene regulation; expresses
            3×FLAG-NLS-zCas9-NLS, gRNA scaffold for insertion of target sequence
            (OsU3 promoter), Bar resistance.
ACCESSION   .
VERSION     .
KEYWORDS    pBUN411
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13518)
  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 13518)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..13518
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..25
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     promoter        164..543
                     /label=OsU3 promoter
                     /note="Oryza sativa (rice) snRNA U3 promoter"
     CDS             823..1614
                     /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        1766..1841
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             4186..4251
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4258..4278
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4303..8403
                     /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             8316..8324
                     /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             8404..8451
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     terminator      8496..8748
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(8757..8774)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8757..8773)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8766..8788)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        8973..9649
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     9586..9606
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             9694..10245
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     polyA_signal    10252..10426
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    10576..10600
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(10848..10871)
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     terminator      10879..10906
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      10998..11084
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      11253..11688
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     rep_origin      complement(11803..12391)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11883..11902)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(12481..13290)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     13204..13223
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     13282..13301
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 gtttacccgc caatatatcc tgtcaaacac tgatagttta aactgaaggc gggaaacgac
       61 aatctgatcc aagctcaagc taagctcacg tgacggaatt aagcttagta attcatccag
      121 gtctccaagt tctaggattt tcagaactgc aacttatttt atcaaggaat ctttaaacat
      181 acgaacagat cacttaaagt tcttctgaag caacttaaag ttatcaggca tgcatggatc
      241 ttggaggaat cagatgtgca gtcagggacc atagcacaag acaggcgtct tctactggtg
      301 ctaccagcaa atgctggaag ccgggaacac tgggtacgtt ggaaaccacg tgatgtgaag
      361 aagtaagata aactgtagga gaaaagcatt tcgtagtggg ccatgaagcc tttcaggaca
      421 tgtattgcag tatgggccgg cccattacgc aattggacga caacaaagac tagtattagt
      481 accacctcgg ctatccacat agatcaaagc tgatttaaaa gagttgtgca gatgatccgt
      541 ggcgtgagac caacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag
      601 cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca
      661 gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca
      721 tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat
      781 gttacgcagc agggcagtcg ccctaaaaca aagttaaaca tcatggggga agcggtgatc
      841 gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg
      901 acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt
      961 gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg
     1021 atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta
     1081 gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa
     1141 ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg
     1201 atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta
     1261 ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta
     1321 aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta
     1381 gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat
     1441 gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa
     1501 gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg
     1561 gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataatgtcta
     1621 gctagaaatt cgttcaagcc gacgccgctt cgcggcgcgg cttaactcaa gcgttagatg
     1681 cactaagcac ataattgctc acagccaaac tatcaggtca agtctgcttt tattattttt
     1741 aagcgtgcat aataagccgg tctcggtttt agagctagaa atagcaagtt aaaataaggc
     1801 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cttttttttt tcgttttgca
     1861 ttgagttttc tccgtcgcat gtttgcagtt ttattttccg ttttgcattg aaatttctcc
     1921 gtctcatgtt tgcagcgtgt tcaaaaagta cgcagctgta tttcacttat ttacggcgcc
     1981 acattttcat gccgtttgtg ccaactatcc cgagctagtg aatacagctt ggcttcacac
     2041 aacactggtg acccgctgac ctgctcgtac ctcgtaccgt cgtacggcac agcatttgga
     2101 attaaagggt gtgatcgata ctgcttgctg ctaagcttga tatcgaattc ctgcagtgca
     2161 gcgtgacccg gtcgtgcccc tctctagaga taatgagcat tgcatgtcta agttataaaa
     2221 aattaccaca tatttttttt gtcacacttg tttgaagtgc agtttatcta tctttataca
     2281 tatatttaaa ctttactcta cgaataatat aatctatagt actacaataa tatcagtgtt
     2341 ttagagaatc atataaatga acagttagac atggtctaaa ggacaattga gtattttgac
     2401 aacaggactc tacagtttta tctttttagt gtgcatgtgt tctccttttt ttttgcaaat
     2461 agcttcacct atataatact tcatccattt tattagtaca tccatttagg gtttagggtt
     2521 aatggttttt atagactaat ttttttagta catctatttt attctatttt agcctctaaa
     2581 ttaagaaaac taaaactcta ttttagtttt tttatttaat aatttagata taaaatagaa
     2641 taaaataaag tgactaaaaa ttaaacaaat accctttaag aaattaaaaa aactaaggaa
     2701 acatttttct tgtttcgagt agataatgcc agcctgttaa acgccgtcga cgagtctaac
     2761 ggacaccaac cagcgaacca gcagcgtcgc gtcgggccaa gcgaagcaga cggcacggca
     2821 tctctgtcgc tgcctctgga cccctctcga gagttccgct ccaccctctc gagagttccg
     2881 ctccaccgtt ggacttgcgt ggcggagcgg cagacgtgag ccggcacggc aggcggcctc
     2941 ctcctcctct cacggcaccg gcagctacgg gggattcctt tcccaccgct ccttcgcttt
     3001 cccttcctcg cccgccgtaa taaatagaca ccccctccac accctctttc cccaacctcg
     3061 tgttgttcgg agcgcacaca cacacaacca gatctccccc aaatccaccc gtcggcacct
     3121 ccgcttcaag gtacgccgct cgtcctcccc ccccccccct ctctaccttc tctagatcgg
     3181 cgttccggtc catggttagg gcccggtagt tctacttctg ttcatgtttg tgttagatcc
     3241 gtgtttgtgt tagatccgtg ctgctagcgt tcgtacacgg atgcgacctg tacgtcagac
     3301 acgttctgat tgctaacttg ccagtgtttc tctttgggga atcctgggat ggctctagcc
     3361 gttccgcaga cgggatcgat ttcatgattt tttttgtttc gttgcatagg gtttggtttg
     3421 cccttttcct ttatttcaat atatgccgtg cacttgtttg tcgggtcatc ttttcatgct
     3481 tttttttgtc ttggttgtga tgatgtggtc tggttgggcg gtcgttctag atcggagtag
     3541 aattctgttt caaactacct ggtggattta ttaattttgg atctgtatgt gtgtgccata
     3601 catattcata gttacgaatt gaagatgatg gatggaaata tcgatctagg ataggtatac
     3661 atgttgatgc gggttttact gatgcatata cagagatgct ttttgttcgc ttggttgtga
     3721 tgatgtggtg tggttgggcg gtcgttcatt cgttctagat cggagtagaa tactgtttca
     3781 aactacctgg tgtatttatt aattttggaa ctgtatgtgt gtgtcataca tcttcatagt
     3841 tacgagttta agatggatgg aaatatcgat ctaggatagg tatacatgtt gatgtgggtt
     3901 ttactgatgc atatacatga tggcatatgc agcatctatt catatgctct aaccttgagt
     3961 acctatctat tataataaac aagtatgttt tataattatt ttgatcttga tatacttgga
     4021 tgatggcata tgcagcagct atatgtggat ttttttagcc ctgccttcat acgctattta
     4081 tttgcttggt actgtttctt ttgtcgatgc tcaccctgtt gtttggtgtt acttctgcag
     4141 cccgggggat ccccaatact tgtatggccg cggccgctct agatggatta caaggaccac
     4201 gacggggatt acaaggacca cgacattgat tacaaggatg atgatgacaa gatggctccg
     4261 aagaagaaga ggaaggttgg catccacggg gtgccagctg ctgacaagaa gtactcgatc
     4321 ggcctcgata ttgggactaa ctctgttggc tgggccgtga tcaccgacga gtacaaggtg
     4381 ccctcaaaga agttcaaggt cctgggcaac accgatcggc attccatcaa gaagaatctc
     4441 attggcgctc tcctgttcga cagcggcgag acggctgagg ctacgcggct caagcgcacc
     4501 gcccgcaggc ggtacacgcg caggaagaat cgcatctgct acctgcagga gattttctcc
     4561 aacgagatgg cgaaggttga cgattctttc ttccacaggc tggaggagtc attcctcgtg
     4621 gaggaggata agaagcacga gcggcatcca atcttcggca acattgtcga cgaggttgcc
     4681 taccacgaga agtaccctac gatctaccat ctgcggaaga agctcgtgga ctccacagat
     4741 aaggcggacc tccgcctgat ctacctcgct ctggcccaca tgattaagtt caggggccat
     4801 ttcctgatcg agggggatct caacccggac aatagcgatg ttgacaagct gttcatccag
     4861 ctcgtgcaga cgtacaacca gctcttcgag gagaacccca ttaatgcgtc aggcgtcgac
     4921 gcgaaggcta tcctgtccgc taggctctcg aagtctcggc gcctcgagaa cctgatcgcc
     4981 cagctgccgg gcgagaagaa gaacggcctg ttcgggaatc tcattgcgct cagcctgggg
     5041 ctcacgccca acttcaagtc gaatttcgat ctcgctgagg acgccaagct gcagctctcc
     5101 aaggacacat acgacgatga cctggataac ctcctggccc agatcggcga tcagtacgcg
     5161 gacctgttcc tcgctgccaa gaatctgtcg gacgccatcc tcctgtctga tattctcagg
     5221 gtgaacaccg agattacgaa ggctccgctc tcagcctcca tgatcaagcg ctacgacgag
     5281 caccatcagg atctgaccct cctgaaggcg ctggtcaggc agcagctccc cgagaagtac
     5341 aaggagatct tcttcgatca gtcgaagaac ggctacgctg ggtacattga cggcggggcc
     5401 tctcaggagg agttctacaa gttcatcaag ccgattctgg agaagatgga cggcacggag
     5461 gagctgctgg tgaagctcaa tcgcgaggac ctcctgagga agcagcggac attcgataac
     5521 ggcagcatcc cacaccagat tcatctcggg gagctgcacg ctatcctgag gaggcaggag
     5581 gacttctacc ctttcctcaa ggataaccgc gagaagatcg agaagattct gactttcagg
     5641 atcccgtact acgtcggccc actcgctagg ggcaactccc gcttcgcttg gatgacccgc
     5701 aagtcagagg agacgatcac gccgtggaac ttcgaggagg tggtcgacaa gggcgctagc
     5761 gctcagtcgt tcatcgagag gatgacgaat ttcgacaaga acctgccaaa tgagaaggtg
     5821 ctccctaagc actcgctcct gtacgagtac ttcacagtct acaacgagct gactaaggtg
     5881 aagtatgtga ccgagggcat gaggaagccg gctttcctgt ctggggagca gaagaaggcc
     5941 atcgtggacc tcctgttcaa gaccaaccgg aaggtcacgg ttaagcagct caaggaggac
     6001 tacttcaaga agattgagtg cttcgattcg gtcgagatct ctggcgttga ggaccgcttc
     6061 aacgcctccc tggggaccta ccacgatctc ctgaagatca ttaaggataa ggacttcctg
     6121 gacaacgagg agaatgagga tatcctcgag gacattgtgc tgacactcac tctgttcgag
     6181 gaccgggaga tgatcgagga gcgcctgaag acttacgccc atctcttcga tgacaaggtc
     6241 atgaagcagc tcaagaggag gaggtacacc ggctggggga ggctgagcag gaagctcatc
     6301 aacggcattc gggacaagca gtccgggaag acgatcctcg acttcctgaa gagcgatggc
     6361 ttcgcgaacc gcaatttcat gcagctgatt cacgatgaca gcctcacatt caaggaggat
     6421 atccagaagg ctcaggtgag cggccagggg gactcgctgc acgagcatat cgcgaacctc
     6481 gctggctcgc cagctatcaa gaaggggatt ctgcagaccg tgaaggttgt ggacgagctg
     6541 gtgaaggtca tgggcaggca caagcctgag aacatcgtca ttgagatggc ccgggagaat
     6601 cagaccacgc agaagggcca gaagaactca cgcgagagga tgaagaggat cgaggagggc
     6661 attaaggagc tggggtccca gatcctcaag gagcacccgg tggagaacac gcagctgcag
     6721 aatgagaagc tctacctgta ctacctccag aatggccgcg atatgtatgt ggaccaggag
     6781 ctggatatta acaggctcag cgattacgac gtcgatcata tcgttccaca gtcattcctg
     6841 aaggatgact ccattgacaa caaggtcctc accaggtcgg acaagaaccg gggcaagtct
     6901 gataatgttc cttcagagga ggtcgttaag aagatgaaga actactggcg ccagctcctg
     6961 aatgccaagc tgatcacgca gcggaagttc gataacctca caaaggctga gaggggcggg
     7021 ctctctgagc tggacaaggc gggcttcatc aagaggcagc tggtcgagac acggcagatc
     7081 actaagcacg ttgcgcagat tctcgactca cggatgaaca ctaagtacga tgagaatgac
     7141 aagctgatcc gcgaggtgaa ggtcatcacc ctgaagtcaa agctcgtctc cgacttcagg
     7201 aaggatttcc agttctacaa ggttcgggag atcaacaatt accaccatgc ccatgacgcg
     7261 tacctgaacg cggtggtcgg cacagctctg atcaagaagt acccaaagct cgagagcgag
     7321 ttcgtgtacg gggactacaa ggtttacgat gtgaggaaga tgatcgccaa gtcggagcag
     7381 gagattggca aggctaccgc caagtacttc ttctactcta acattatgaa tttcttcaag
     7441 acagagatca ctctggccaa tggcgagatc cggaagcgcc ccctcatcga gacgaacggc
     7501 gagacggggg agatcgtgtg ggacaagggc agggatttcg cgaccgtcag gaaggttctc
     7561 tccatgccac aagtgaatat cgtcaagaag acagaggtcc agactggcgg gttctctaag
     7621 gagtcaattc tgcctaagcg gaacagcgac aagctcatcg cccgcaagaa ggactgggat
     7681 ccgaagaagt acggcgggtt cgacagcccc actgtggcct actcggtcct ggttgtggcg
     7741 aaggttgaga agggcaagtc caagaagctc aagagcgtga aggagctgct ggggatcacg
     7801 attatggagc gctccagctt cgagaagaac ccgatcgatt tcctggaggc gaagggctac
     7861 aaggaggtga agaaggacct gatcattaag ctccccaagt actcactctt cgagctggag
     7921 aacggcagga agcggatgct ggcttccgct ggcgagctgc agaaggggaa cgagctggct
     7981 ctgccgtcca agtatgtgaa cttcctctac ctggcctccc actacgagaa gctcaagggc
     8041 agccccgagg acaacgagca gaagcagctg ttcgtcgagc agcacaagca ttacctcgac
     8101 gagatcattg agcagatttc cgagttctcc aagcgcgtga tcctggccga cgcgaatctg
     8161 gataaggtcc tctccgcgta caacaagcac cgcgacaagc caatcaggga gcaggctgag
     8221 aatatcattc atctcttcac cctgacgaac ctcggcgccc ctgctgcttt caagtacttc
     8281 gacacaacta tcgatcgcaa gaggtacaca agcactaagg aggtcctgga cgcgaccctc
     8341 atccaccagt cgattaccgg cctctacgag acgcgcatcg acctgtctca gctcgggggc
     8401 gacaagcggc cagcggcgac gaagaaggcg gggcaggcga agaagaagaa gtgagctcag
     8461 gcctccatac aagtattggg gatccgaatt tccccgatcg ttcaaacatt tggcaataaa
     8521 gtttcttaag attgaatcct gttgccggtc ttgcgatgat tatcatataa tttctgttga
     8581 attacgttaa gcatgtaata attaacatgt aatgcatgac gttatttatg agatgggttt
     8641 ttatgattag agtcccgcaa ttatacattt aatacgcgat agaaaacaaa atatagcgcg
     8701 caaactagga taaattatcg cgcgcggtgt catctatgtt actagatcgg gaattcactg
     8761 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     8821 gcagcacatc cccctttcgc caggggtacc gagctctaga attcgaggcg gtttgcgtat
     8881 tggctagagc agcttgccaa catggtggag cacgacactc tcgtctactc caagaatatc
     8941 aaagatacag tctcagaaga ccaaagggct attgagactt ttcaacaaag ggtaatatcg
     9001 ggaaacctcc tcggattcca ttgcccagct atctgtcact tcatcaaaag gacagtagaa
     9061 aaggaaggtg gcacctacaa atgccatcat tgcgataaag gaaaggctat cgttcaagat
     9121 gcctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat cgtggaaaaa
     9181 gaagacgttc caaccacgtc ttcaaagcaa gtggattgat gtgaacatgg tggagcacga
     9241 cactctcgtc tactccaaga atatcaaaga tacagtctca gaagaccaaa gggctattga
     9301 gacttttcaa caaagggtaa tatcgggaaa cctcctcgga ttccattgcc cagctatctg
     9361 tcacttcatc aaaaggacag tagaaaagga aggtggcacc tacaaatgcc atcattgcga
     9421 taaaggaaag gctatcgttc aagatgcctc tgccgacagt ggtcccaaag atggaccccc
     9481 acccacgagg agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga
     9541 ttgatgtgat atctccactg acgtaaggga tgacgcacaa tcccactatc cttcgcaaga
     9601 cccttcctct atataaggaa gttcatttca tttggagagg acacgctgaa atcaccagtc
     9661 tctctctaca aatctatctc tctcgagtct accatgagcc cagaacgacg cccggccgac
     9721 atccgccgtg ccaccgaggc ggacatgccg gcggtctgca ccatcgtcaa ccactacatc
     9781 gagacaagca cggtcaactt ccgtaccgag ccgcaggaac cgcaggagtg gacggacgac
     9841 ctcgtccgtc tgcgggagcg ctatccctgg ctcgtcgccg aggtggacgg cgaggtcgcc
     9901 ggcatcgcct acgcgggccc ctggaaggca cgcaacgcct acgactggac ggccgagtcg
     9961 accgtgtacg tctccccccg ccaccagcgg acgggactgg gctccacgct ctacacccac
    10021 ctgctgaagt ccctggaggc acagggcttc aagagcgtgg tcgctgtcat cgggctgccc
    10081 aacgacccga gcgtgcgcat gcacgaggcg ctcggatatg ccccccgcgg catgctgcgg
    10141 gcggccggct tcaagcacgg gaactggcat gacgtgggtt tctggcagct ggacttcagc
    10201 ctgccggtac cgccccgtcc ggtcctgccc gtcaccgaga tttgactcga gtttctccat
    10261 aataatgtgt gagtagttcc cagataaggg aattagggtt cctatagggt ttcgctcatg
    10321 tgttgagcat ataagaaacc cttagtatgt atttgtattt gtaaaatact tctatcaata
    10381 aaatttctaa ttcctaaaac caaaatccag tactaaaatc cagatccccc gaattaattc
    10441 ggcgttaatt cagtacatta aaaacgtccg caatgtgtta ttaagttgtc actagtcagg
    10501 ttaactcaat tcggcgttaa ttcagtacat taaaaacgtc cgcaatgtgt tattaagttg
    10561 tctaagcgtc aatttgttta caccacaata tatcctgcca ccagccagcc aacagctccc
    10621 cgaccggcag ctcggcacaa aatcaccact cgatacaggc agcccatcag tccgggacgg
    10681 cgtcagcggg agagccgttg taaggcggca gactttgctc atgttaccga tgctattcgg
    10741 aagaacggca actaagctgc cgggtttgaa acacggatga tctcgcggag ggtagcatgt
    10801 tgattgtaac gatgacagag cgttgctgcc tgtgatcact taagtaacta actaacagga
    10861 agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc
    10921 ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc
    10981 aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata
    11041 aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc
    11101 tactctcgct tagtagttag acgtccccga gatccatgct agaccatgaa tccagaagcc
    11161 cgagaggttg ccgcctttcg ggctttttct ttttcaaaaa aaaaaattta taaaacgatc
    11221 tgttgcggcc ggccgccggg ttgtgggcaa aggcgctcga cggtgggcaa ccgcttgcgg
    11281 ttgtccacgg gcggagccgg tgcgcgtagc gcattgtcca caagccaagg gcgaccaata
    11341 attgatatat atattcataa ttgaaaagct aattgaacat actacttgct gtaactactt
    11401 gccggagcga ggggtgtttg caagctgttg atctgaaagg gctattagcg ttctcacgtg
    11461 cctttttgat tagcgatttc acgtgacctt attagcgatt tcacgtactc cgattagcga
    11521 tttcacgtac cctgattagc gatttcacgt ggatagtttt tggagcgggc cggaaagccc
    11581 cgtgaatcaa ggctttgcgg ggcattagcg gtttcacgtg gataactacc ctctatccac
    11641 aggcttccgg ggataaaaaa gcccgctcga cggcgggctg ttggatgggg atctagcggt
    11701 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
    11761 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
    11821 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
    11881 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
    11941 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
    12001 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
    12061 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
    12121 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
    12181 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
    12241 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
    12301 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
    12361 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
    12421 tgacgctcag tggaacgggg cccaatctga ataatgttac aaccaattaa ccaattctga
    12481 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
    12541 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
    12601 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
    12661 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
    12721 tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca
    12781 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
    12841 cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
    12901 atgcaaccgg cgcagggaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
    12961 ttcttctaat acctggaatg ctgtttttcc ggggatcgca gtggtgagta accatgcatc
    13021 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
    13081 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
    13141 caactctggc gcatcgggct tcccatacaa gcgatagatt gtcgcacctg attgcccgac
    13201 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
    13261 cctcgacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc
    13321 agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt
    13381 ttgagacacg ggccagagct gcagtttgat cccgagggga accctgtggt tgacatgcac
    13441 atacaaatgg acgaacggat aaaccttttc acgccctttt aaatatccgt tattctaata
    13501 aacgctcttt tctcttag
//