pG3B-U3Ub

LOCUS       pG3B-U3Ub               13821 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  pGreen3 CRISPR/Cas9 .
ACCESSION   .
VERSION     .
KEYWORDS    pG3B-U3Ub
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13821)
  AUTHORS   Zhang Q, Zhang Y, Lu MH, Chai YP, Jiang YY, Zhou Y, Wang XC, Chen QJ
  TITLE     A Novel Ternary Vector System United with Morphogenic Genes Enhances
            CRISPR/Cas Delivery in Maize.
  JOURNAL   Plant Physiol. 2019 Sep 26. pii: pp.19.00767. doi: 
            10.1104/pp.19.00767.
  PUBMED    31558579
REFERENCE   2  (bases 1 to 13821)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..13821
                     /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"
     primer_bind     213..235
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     227..244
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     228..244
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        311..690
                     /label=OsU3 promoter
                     /note="Oryza sativa (rice) snRNA U3 promoter"
     promoter        713..817
                     /gene="bla"
                     /label=AmpR promoter
     CDS             818..1678
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1036..1055)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     misc_RNA        1827..1902
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        1968..3960
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     CDS             3979..4044
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4051..4071
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4096..8196
                     /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             8109..8117
                     /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             8197..8244
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(8901..8917)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8901..8917)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8914..8936)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8925..8941
                     /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(8949..8979)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8994..9015
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9206..9882
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     9819..9839
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             9927..10478
                     /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    10485..10659
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    10737..10761
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(11139..11158)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             11150..11959
                     /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     complement(11217..11236)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      12038..12124
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      12216..12243
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      12322..12910
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12811..12830
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtttacccgc caatatatcc tgtcaaacac tgatagttta aactgaaggc gggaaacgac
       61 aatctgatcc aagctcaagc tgctctagca ttcgccattc aggctgcgca actgttggga
      121 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
      181 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
      241 cagtgccaag cttagtaatt catccaggtc accaagttct aggattttca gaactgcaac
      301 ttattttatc aaggaatctt taaacatacg aacagatcac ttaaagttct tctgaagcaa
      361 cttaaagtta tcaggcttgc atggatcttg gaggaatcag atgtgcagtc agggaccata
      421 gcacaagaca ggcgtcttct actggtgcta ccagcaaatg ctggaagccg ggaacactgg
      481 gtacgttgga aaccacgtga tgtgaagaag taagataaac tgtaggagaa aagcatttcg
      541 tagtgggcca tgaagccttt caggacatgt attgcagtat gggccggccc attacgcaat
      601 tggacgacaa caaagtctag tattagtacc acctcggcta tccacataga tcaaagctga
      661 tttaaaagag ttgtgcagat gatccgtggc gtgagacctt cggggaaatg tgcgcggaac
      721 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatgg gacaataacc
      781 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
      841 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
      901 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
      961 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     1021 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     1081 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     1141 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     1201 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     1261 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     1321 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     1381 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     1441 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     1501 tattgctgat aaatctggag ccggtgagcg tggctctcgc ggtatcattg cagcactggg
     1561 gccagatggt aagccctccc gtatcgtagt tatctacacc acggggagtc aggcaactat
     1621 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     1681 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     1741 aaggatctag gtgaagatcc tttttgataa tcccatgacc aaaatccctt aacgtgagtt
     1801 ttcgttccac tgagcgtcag gtctcagttt tagagctaga aatagcaagt taaaataagg
     1861 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctttttttt ttgtccttct
     1921 gtttttttag tcagtctctt ttttcagaag tacaacatct tgcatgcctg cagtgcagcg
     1981 tgacccggtc gtgcccctct ctagagataa tgagcattgc atgtctaagt tataaaaaat
     2041 taccacatat tttttttgtc acacttgttt gaagtgcagt ttatctatct ttatacatat
     2101 atttaaactt tactctacga ataatataat ctatagtact acaataatat cagtgtttta
     2161 gagaatcata taaatgaaca gttagacatg gtctaaagga caattgagta ttttgacaac
     2221 aggactctac agttttatct ttttagtgtg catgtgttct cctttttttt tgcaaatagc
     2281 ttcacctata taatacttca tccattttat tagtacatcc atttagggtt tagggttaat
     2341 ggtttttata gactaatttt tttagtacat ctattttatt ctattttagc ctctaaatta
     2401 agaaaactaa aactctattt tagttttttt atttaataat ttagatataa aatagaataa
     2461 aataaagtga ctaaaaatta aacaaatacc ctttaagaaa ttaaaaaaac taaggaaaca
     2521 tttttcttgt ttcgagtaga taatgccagc ctgttaaacg ccgtcgacga gtctaacgga
     2581 caccaaccag cgaaccagca gcgtcgcgtc gggccaagcg aagcagacgg cacggcatct
     2641 ctgtcgctgc ctctggaccc ctctcgagag ttccgctcca ccgttggact tgctccgctg
     2701 tcggcatcca gaaattgcgt ggcggagcgg cagacgtgag ccggcacggc aggcggcctc
     2761 ctcctcctct cacggcaccg gcagctacgg gggattcctt tcccaccgct ccttcgcttt
     2821 cccttcctcg cccgccgtaa taaatagaca ccccctccac accctctttc cccaacctcg
     2881 tgttgttcgg agcgcacaca cacacaacca gatctccccc aaatccaccc gtcggcacct
     2941 ccgcttcaag gtacgccgct cgtcctcccc ccccccccct ctctaccttc tctagatcgg
     3001 cgttccggtc catggttagg gcccggtagt tctacttctg ttcatgtttg tgttagatcc
     3061 gtgtttgtgt tagatccgtg ctgctagcgt tcgtacacgg atgcgacctg tacgtcagac
     3121 acgttctgat tgctaacttg ccagtgtttc tctttgggga atcctgggat ggctctagcc
     3181 gttccgcaga cgggatcgat ttcatgattt tttttgtttc gttgcatagg gtttggtttg
     3241 cccttttcct ttatttcaat atatgccgtg cacttgtttg tcgggtcatc ttttcatgct
     3301 tttttttgtc ttggttgtga tgatgtggtc tggttgggcg gtcgttctag atcggagtag
     3361 aattctgttt caaactacct ggtggattta ttaattttgg atctgtatgt gtgtgccata
     3421 catattcata gttacgaatt gaagatgatg gatggaaata tcgatctagg ataggtatac
     3481 atgttgatgc gggttttact gatgcatata cagagatgct ttttgttcgc ttggttgtga
     3541 tgatgtggtg tggttgggcg gtcgttcatt cgttctagat cggagtagaa tactgtttca
     3601 aactacctgg tgtatttatt aattttggaa ctgtatgtgt gtgtcataca tcttcatagt
     3661 tacgagttta agatggatgg aaatatcgat ctaggatagg tatacatgtt gatgtgggtt
     3721 ttactgatgc atatacatga tggcatatgc agcatctatt catatgctct aaccttgagt
     3781 acctatctat tataataaac aagtatgttt tataattatt ttgatcttga tatacttgga
     3841 tgatggcata tgcagcagct atatgtggat ttttttagcc ctgccttcat acgctattta
     3901 tttgcttggt actgtttctt ttgtcgatgc tcaccctgtt gtttggtgtt acttctgcag
     3961 ccctaggcct actagatgga ttacaaggac cacgacgggg attacaagga ccacgacatt
     4021 gattacaagg atgatgatga caagatggct ccgaagaaga agaggaaggt tggcatccac
     4081 ggggtgccag ctgctgacaa gaagtactcg atcggcctcg atattgggac taactctgtt
     4141 ggctgggccg tgatcaccga cgagtacaag gtgccctcaa agaagttcaa ggtcctgggc
     4201 aacaccgatc ggcattccat caagaagaat ctcattggcg ctctcctgtt cgacagcggc
     4261 gagacggctg aggctacgcg gctcaagcgc accgcccgca ggcggtacac gcgcaggaag
     4321 aatcgcatct gctacctgca ggagattttc tccaacgaga tggcgaaggt tgacgattct
     4381 ttcttccaca ggctggagga gtcattcctc gtggaggagg ataagaagca cgagcggcat
     4441 ccaatcttcg gcaacattgt cgacgaggtt gcctaccacg agaagtaccc tacgatctac
     4501 catctgcgga agaagctcgt ggactccaca gataaggcgg acctccgcct gatctacctc
     4561 gctctggccc acatgattaa gttcaggggc catttcctga tcgaggggga tctcaacccg
     4621 gacaatagcg atgttgacaa gctgttcatc cagctcgtgc agacgtacaa ccagctcttc
     4681 gaggagaacc ccattaatgc gtcaggcgtc gacgcgaagg ctatcctgtc cgctaggctc
     4741 tcgaagtctc ggcgcctcga gaacctgatc gcccagctgc cgggcgagaa gaagaacggc
     4801 ctgttcggga atctcattgc gctcagcctg gggctcacgc ccaacttcaa gtcgaatttc
     4861 gatctcgctg aggacgccaa gctgcagctc tccaaggaca catacgacga tgacctggat
     4921 aacctcctgg cccagatcgg cgatcagtac gcggacctgt tcctcgctgc caagaatctg
     4981 tcggacgcca tcctcctgtc tgatattctc agggtgaaca ccgagattac gaaggctccg
     5041 ctctcagcct ccatgatcaa gcgctacgac gagcaccatc aggatctgac cctcctgaag
     5101 gcgctggtca ggcagcagct ccccgagaag tacaaggaga tcttcttcga tcagtcgaag
     5161 aacggctacg ctgggtacat tgacggcggg gcctctcagg aggagttcta caagttcatc
     5221 aagccgattc tggagaagat ggacggcacg gaggagctgc tggtgaagct caatcgcgag
     5281 gacctcctga ggaagcagcg gacattcgat aacggcagca tcccacacca gattcatctc
     5341 ggggagctgc acgctatcct gaggaggcag gaggacttct accctttcct caaggataac
     5401 cgcgagaaga tcgagaagat tctgactttc aggatcccgt actacgtcgg cccactcgct
     5461 aggggcaact cccgcttcgc ttggatgacc cgcaagtcag aggagacgat cacgccgtgg
     5521 aacttcgagg aggtggtcga caagggcgct agcgctcagt cgttcatcga gaggatgacg
     5581 aatttcgaca agaacctgcc aaatgagaag gtgctcccta agcactcgct cctgtacgag
     5641 tacttcacag tctacaacga gctgactaag gtgaagtatg tgaccgaggg catgaggaag
     5701 ccggctttcc tgtctgggga gcagaagaag gccatcgtgg acctcctgtt caagaccaac
     5761 cggaaggtca cggttaagca gctcaaggag gactacttca agaagattga gtgcttcgat
     5821 tcggtcgaga tctctggcgt tgaggaccgc ttcaacgcct ccctggggac ctaccacgat
     5881 ctcctgaaga tcattaagga taaggacttc ctggacaacg aggagaatga ggatatcctc
     5941 gaggacattg tgctgacact cactctgttc gaggaccggg agatgatcga ggagcgcctg
     6001 aagacttacg cccatctctt cgatgacaag gtcatgaagc agctcaagag gaggaggtac
     6061 accggctggg ggaggctgag caggaagctc atcaacggca ttcgggacaa gcagtccggg
     6121 aagacgatcc tcgacttcct gaagagcgat ggcttcgcga accgcaattt catgcagctg
     6181 attcacgatg acagcctcac attcaaggag gatatccaga aggctcaggt gagcggccag
     6241 ggggactcgc tgcacgagca tatcgcgaac ctcgctggct cgccagctat caagaagggg
     6301 attctgcaga ccgtgaaggt tgtggacgag ctggtgaagg tcatgggcag gcacaagcct
     6361 gagaacatcg tcattgagat ggcccgggag aatcagacca cgcagaaggg ccagaagaac
     6421 tcacgcgaga ggatgaagag gatcgaggag ggcattaagg agctggggtc ccagatcctc
     6481 aaggagcacc cggtggagaa cacgcagctg cagaatgaga agctctacct gtactacctc
     6541 cagaatggcc gcgatatgta tgtggaccag gagctggata ttaacaggct cagcgattac
     6601 gacgtcgatc atatcgttcc acagtcattc ctgaaggatg actccattga caacaaggtc
     6661 ctcaccaggt cggacaagaa ccggggcaag tctgataatg ttccttcaga ggaggtcgtt
     6721 aagaagatga agaactactg gcgccagctc ctgaatgcca agctgatcac gcagcggaag
     6781 ttcgataacc tcacaaaggc tgagaggggc gggctctctg agctggacaa ggcgggcttc
     6841 atcaagaggc agctggtcga gacacggcag atcactaagc acgttgcgca gattctcgac
     6901 tcacggatga acactaagta cgatgagaat gacaagctga tccgcgaggt gaaggtcatc
     6961 accctgaagt caaagctcgt ctccgacttc aggaaggatt tccagttcta caaggttcgg
     7021 gagatcaaca attaccacca tgcccatgac gcgtacctga acgcggtggt cggcacagct
     7081 ctgatcaaga agtacccaaa gctcgagagc gagttcgtgt acggggacta caaggtttac
     7141 gatgtgagga agatgatcgc caagtcggag caggagattg gcaaggctac cgccaagtac
     7201 ttcttctact ctaacattat gaatttcttc aagacagaga tcactctggc caatggcgag
     7261 atccggaagc gccccctcat cgagacgaac ggcgagacgg gggagatcgt gtgggacaag
     7321 ggcagggatt tcgcgaccgt caggaaggtt ctctccatgc cacaagtgaa tatcgtcaag
     7381 aagacagagg tccagactgg cgggttctct aaggagtcaa ttctgcctaa gcggaacagc
     7441 gacaagctca tcgcccgcaa gaaggactgg gatccgaaga agtacggcgg gttcgacagc
     7501 cccactgtgg cctactcggt cctggttgtg gcgaaggttg agaagggcaa gtccaagaag
     7561 ctcaagagcg tgaaggagct gctggggatc acgattatgg agcgctccag cttcgagaag
     7621 aacccgatcg atttcctgga ggcgaagggc tacaaggagg tgaagaagga cctgatcatt
     7681 aagctcccca agtactcact cttcgagctg gagaacggca ggaagcggat gctggcttcc
     7741 gctggcgagc tgcagaaggg gaacgagctg gctctgccgt ccaagtatgt gaacttcctc
     7801 tacctggcct cccactacga gaagctcaag ggcagccccg aggacaacga gcagaagcag
     7861 ctgttcgtcg agcagcacaa gcattacctc gacgagatca ttgagcagat ttccgagttc
     7921 tccaagcgcg tgatcctggc cgacgcgaat ctggataagg tcctctccgc gtacaacaag
     7981 caccgcgaca agccaatcag ggagcaggct gagaatatca ttcatctctt caccctgacg
     8041 aacctcggcg cccctgctgc tttcaagtac ttcgacacaa ctatcgatcg caagaggtac
     8101 acaagcacta aggaggtcct ggacgcgacc ctcatccacc agtcgattac cggcctctac
     8161 gagacgcgca tcgacctgtc tcagctcggg ggcgacaagc ggccagcggc gacgaagaag
     8221 gcggggcagg cgaagaagaa gaagtgagct cagagctttc gttcgtatca tcggtttcga
     8281 caacgttcgt caagttcaat gcatcagttt cattgcgcac acaccagaat cctactgagt
     8341 ttgagtatta tggcattggg aaaactgttt ttcttgtacc atttgttgtg cttgtaattt
     8401 actgtgtttt ttattcggtt ttcgctatcg aactgtgaaa tggaaatgga tggagaagag
     8461 ttaatgaatg atatggtcct tttgttcatt ctcaaattaa tattatttgt tttttctctt
     8521 atttgttgtg tgttgaattt gaaattataa gagatatgca aacattttgt tttgagtaaa
     8581 aatgtgtcaa atcgtggcct ctaatgaccg aagttaatat gaggagtaaa acacttgtag
     8641 ttgtaccatt atgcttattc actaggcaac aaatatattt tcagacctag aaaagctgca
     8701 aatgttactg aatacaagta tgtcctcttg tgttttagac atttatgaac tttcctttat
     8761 gtaattttcc agaatccttg tcagattcta atcattgctt tataattata gttatactca
     8821 tggatttgta gttgagtatg aaaatatttt ttaatgcatt ttatgacttg ccaattgatt
     8881 gacaacgaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     8941 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     9001 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     9061 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattggctag
     9121 agcagcttgc caacatggtg gagcacgaca ctctcgtcta ctccaagaat atcaaagata
     9181 cagtctcaga agaccaaagg gctattgaga cttttcaaca aagggtaata tcgggaaacc
     9241 tcctcggatt ccattgccca gctatctgtc acttcatcaa aaggacagta gaaaaggaag
     9301 gtggcaccta caaatgccat cattgcgata aaggaaaggc tatcgttcaa gatgcctctg
     9361 ccgacagtgg tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg
     9421 ttccaaccac gtcttcaaag caagtggatt gatgtgaaca tggtggagca cgacactctc
     9481 gtctactcca agaatatcaa agatacagtc tcagaagacc aaagggctat tgagactttt
     9541 caacaaaggg taatatcggg aaacctcctc ggattccatt gcccagctat ctgtcacttc
     9601 atcaaaagga cagtagaaaa ggaaggtggc acctacaaat gccatcattg cgataaagga
     9661 aaggctatcg ttcaagatgc ctctgccgac agtggtccca aagatggacc cccacccacg
     9721 aggagcatcg tggaaaaaga agacgttcca accacgtctt caaagcaagt ggattgatgt
     9781 gatatctcca ctgacgtaag ggatgacgca caatcccact atccttcgca agacccttcc
     9841 tctatataag gaagttcatt tcatttggag aggacacgct gaaatcacca gtctctctct
     9901 acaaatctat ctctctcgag tctaccatga gcccagaacg acgcccggcc gacatccgcc
     9961 gtgccaccga ggcggacatg ccggcggtct gcaccatcgt caaccactac atcgagacaa
    10021 gcacggtcaa cttccgtacc gagccgcagg aaccgcagga gtggacggac gacctcgtcc
    10081 gtctgcggga gcgctatccc tggctcgtcg ccgaggtgga cggcgaggtc gccggcatcg
    10141 cctacgcggg cccctggaag gcacgcaacg cctacgactg gacggccgag tcgaccgtgt
    10201 acgtctcccc ccgccaccag cggacgggac tgggctccac gctctacacc cacctgctga
    10261 agtccctgga ggcacagggc ttcaagagcg tggtcgctgt catcgggctg cccaacgacc
    10321 cgagcgtgcg catgcacgag gcgctcggat atgccccacg cggcatgctg cgggcggccg
    10381 gcttcaagca cgggaactgg catgacgtgg gtttctggca gctggacttc agcctgccgg
    10441 taccgccccg tccggtcctg cccgtcaccg agatttgact cgagtttctc cataataatg
    10501 tgtgagtagt tcccagataa gggaattagg gttcctatag ggtttcgctc atgtgttgag
    10561 catataagaa acccttagta tgtatttgta tttgtaaaat acttctatca ataaaatttc
    10621 taattcctaa aaccaaaatc cagtactaaa atccagatcc cccgaattaa ttcggcgtta
    10681 attcagtaca ttaaaaacgt ccgcaatgtg ttattaagtt gtctaagcgt caatttgttt
    10741 acaccacaat atatcctgcc accagccagc caacagctcc ccgaccggca gctcggcaca
    10801 aaatcaccac tcgatacagg cagcccatca gtccgggacg gcgtcagcgg gagagccgtt
    10861 gtaaggcggc agactttgct catgttaccg atgctattcg gaagaacggc aactaagctg
    10921 ccgggtttga aacacggatg atctcgcgga gggtagcatg ttgattgtaa cgatgacaga
    10981 gcgttgctgc ctgtgatcac cgcggctggc aaacagctat tatgggtatt atgggtctag
    11041 tctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
    11101 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
    11161 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
    11221 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
    11281 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
    11341 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
    11401 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
    11461 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
    11521 gtccctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
    11581 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
    11641 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
    11701 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
    11761 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
    11821 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
    11881 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
    11941 gctcgatgag tttttctaat cagaattggt taattggttg taacattatt cagattggcg
    12001 tctaactact aagcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
    12061 ggaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
    12121 aaatccgccg ggagcggatt tgaacgttgt gaagcaacgg cccggagggt ggcgggcagg
    12181 acgcccgcca taaactgcca ggcatcaaac taagcagaag gccatcctga cggatggcct
    12241 ttttgcgttt ctacaaactc ttcctgttag ggccccgttc cactgagcgt cagaccccgt
    12301 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
    12361 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
    12421 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
    12481 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
    12541 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
    12601 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
    12661 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
    12721 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
    12781 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
    12841 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
    12901 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
    12961 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgctag
    13021 atagctggtt gccctcgccg ctgggctggc ggccgtctat ggccctgcaa acgcgccaga
    13081 aacgccgtcg aagccgtgtg cgagacaccc cggccgccgg cgttgtggat acctcgcgga
    13141 aaacttggcc ctcactgaca gatgaggggc ggacgttgac acttgagggg ccgactcacc
    13201 cggcgcggcg ttgacagatg aggggcaggc tcgatttcgg ccggcgacgt ggagctggcc
    13261 agcctcgcaa atcggcgaaa acgcctgatt ttacgcgagt ttcccacaga tgatgtggac
    13321 aagcctgggg ataagtgccc tgcggtattg acacttgagg ggcgcgacta ctgacagatg
    13381 aggggcgcga tccttgacac ttgaggggca gagtgctgac agatgagggg cgcacctatt
    13441 gacatttgag gggctgtcca caggcagaaa atccagcatt tgcaagggtt tccgcccgtt
    13501 tttcggccac cgctaacctg tcttttaacc tgcttttaaa ccaatattta taaaccttgt
    13561 ttttaaccag ggctgcgccc tgtgcgcgtg accgcgcacg ccgaaggggg gtgccccccc
    13621 ttctcgaacc ctcccggtcg agtcatggtc tagcatggat ctcggggacg tcacccataa
    13681 tacccataat agctgtttgc cagtactttg atcccgaggg gaaccctgtg gttggcatgc
    13741 acatacaaat ggacgaacgg ataaaccttt tcacgccctt ttaaatatcc gattattcta
    13801 ataaacgctc ttttctctta g
//