| 质粒名称: | pSC2 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 91180 |
| 存储实验室: | Daniel Voytas |
|
载体骨架基本信息 |
|
| 载体名称: | pDIRECT_23C |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 植物表达，CRISPR |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | gRNAs targeting NCR genes |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | 未知(未破坏) |
| 3' 克隆位点: | 未知(未破坏) |
| 5' 测序引物 | 未知 |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pSC2 was a gift from Daniel Voytas (Addgene plasmid # 91180 ; http://n2t.net/addgene:91180 ; RRID:Addgene_91180) |
| 参考文献部分: | A multi-purpose toolkit to enable advanced genome engineering in plants. Cermak T, Curtin SJ, Gil-Humanes J, Cegan R, Kono TJY, Konecna E, Belanto JJ, Starker CG, Mathre JW, Greenstein RL, Voytas DF. Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. 10.1105/tpc.16.00922 PubMed 28522548 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pSC2 15703 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION T-DNA vector for combinatorial deletion of NCR genes in medicago
truncatula (Csy4 array of 6 gRNAs under the control of CmYLCV
promoter).
ACCESSION .
VERSION .
KEYWORDS pSC2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 15703)
AUTHORS Cermak T, Curtin SJ, Gil-Humanes J, Cegan R, Kono TJY, Konecna E,
Belanto JJ, Starker CG, Mathre JW, Greenstein RL, Voytas DF
TITLE A multi-purpose toolkit to enable advanced genome engineering in
plants.
JOURNAL Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi:
10.1105/tpc.16.00922.
PUBMED 28522548
REFERENCE 2 (bases 1 to 15703)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..15703
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(291..1085)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 1510..1534
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal 1612..1786
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(1793..2344)
/codon_start=1
/gene="Streptomyces hygroscopicus bar"
/product="phosphinothricin acetyltransferase"
/label=BlpR
/note="confers resistance to bialophos or phosphinothricin"
/translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
WQLDFSLPVPPRPVLPVTEI"
promoter complement(2389..3065)
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
protein_bind 3256..3277
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3292..3322
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3330..3346
/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 3335..3357
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 3354..3370
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3354..3370
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 3900..4245
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
CDS 4837..4893
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 4894..8997
/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 9010..9030
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
protein_bind 9791..9805
/label=Csy4 site
/bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
/note="cleavage site for the Csy4 endoribonuclease
(Haurwitz et al., 2010)"
misc_RNA 9826..9901
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
protein_bind 9907..9921
/label=Csy4 site
/bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
/note="cleavage site for the Csy4 endoribonuclease
(Haurwitz et al., 2010)"
misc_RNA 9942..10017
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
protein_bind 10023..10037
/label=Csy4 site
/bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
/note="cleavage site for the Csy4 endoribonuclease
(Haurwitz et al., 2010)"
misc_RNA 10058..10133
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
protein_bind 10139..10153
/label=Csy4 site
/bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
/note="cleavage site for the Csy4 endoribonuclease
(Haurwitz et al., 2010)"
misc_RNA 10174..10249
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
protein_bind 10255..10269
/label=Csy4 site
/bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
/note="cleavage site for the Csy4 endoribonuclease
(Haurwitz et al., 2010)"
misc_RNA 10290..10365
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
protein_bind 10371..10385
/label=Csy4 site
/bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
/note="cleavage site for the Csy4 endoribonuclease
(Haurwitz et al., 2010)"
misc_RNA 10406..10481
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
protein_bind 10487..10501
/label=Csy4 site
/bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
/note="cleavage site for the Csy4 endoribonuclease
(Haurwitz et al., 2010)"
polyA_signal 10521..10695
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
primer_bind complement(10714..10731)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(10714..10730)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(10723..10745)
/label=M13/pUC Forward
/note="In lacZ gene"
misc_feature 10933..10957
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 12258..12887
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 13316..14389
/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"
rep_origin 14455..14649
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
primer_bind complement(14885..14907)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 14993..15133
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 15048..15067
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(15148..15165)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(15319..204)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(15399..15418)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
61 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
121 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
181 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
241 cagtggaacg aaaactcacg ttaagggatt ttggtcatgc attctaggta ctaaaacaat
301 tcatccagta aaatataata ttttattttc tcccaatcag gcttgatccc cagtaagtca
361 aaaaatagct cgacatactg ttcttccccg atatcctccc tgatcgaccg gacgcagaag
421 gcaatgtcat accacttgtc cgccctgccg cttctcccaa gatcaataaa gccacttact
481 ttgccatctt tcacaaagat gttgctgtct cccaggtcgc cgtgggaaaa gacaagttcc
541 tcttcgggct tttccgtctt taaaaaatca tacagctcgc gcggatcttt aaatggagtg
601 tcttcttccc agttttcgca atccacatcg gccagatcgt tattcagtaa gtaatccaat
661 tcggctaagc ggctgtctaa gctattcgta tagggacaat ccgatatgtc gatggagtga
721 aagagcctga tgcactccgc atacagctcg ataatctttt cagggctttg ttcatcttca
781 tactcttccg agcaaaggac gccatcggcc tcactcatga gcagattgct ccagccatca
841 tgccgttcaa agtgcaggac ctttggaaca ggcagctttc cttccagcca tagcatcatg
901 tccttttccc gttccacatc ataggtggtc cctttatacc ggctgtccgt catttttaaa
961 tataggtttt cattttctcc caccagctta tataccttag caggagacat tccttccgta
1021 tcttttacgc agcggtattt ttcgatcagt tttttcaatt ccggtgatat tctcatttta
1081 gccatttatt atttccttcc tcttttctac agtatttaaa gataccccaa gaagctaatt
1141 ataacaagac gaactccaat tcactgttcc ttgcattcta aaaccttaaa taccagaaaa
1201 cagctttttc aaagttgttt tcaaagttgg cgtataacat agtatcgacg gagccgattt
1261 tgaaaccgcg gtgatcacag gcagcaacgc tctgtcatcg ttacaatcaa catgctaccc
1321 tccgcgagat catccgtgtt tcaaacccgg cagcttagtt gccgttcttc cgaatagcat
1381 cggtaacatg agcaaagtct gccgccttac aacggctctc ccgctgacgc cgtcccggac
1441 tgatgggctg cctgtatcga gtggtgattt tgtgccgagc tgccggtcgg ggagctgttg
1501 gctggctggt ggcaggatat attgtggtgt aaacaaattg acgcttagac aacttaataa
1561 cacattgcgg acgtttttaa tgtactgaat taacgccgaa ttaattcggg ggatctggat
1621 tttagtactg gattttggtt ttaggaatta gaaattttat tgatagaagt attttacaaa
1681 tacaaataca tactaagggt ttcttatatg ctcaacacat gagcgaaacc ctataggaac
1741 cctaattccc ttatctggga actactcaca cattattatg gagaaactcg agtcaaatct
1801 cggtgacggg caggaccgga cggggcggta ccggcaggct gaagtccagc tgccagaaac
1861 ccacgtcatg ccagttcccg tgcttgaagc cggccgcccg cagcatgccg cggggggcat
1921 atccgagcgc ctcgtgcatg cgcacgctcg ggtcgttggg cagcccgatg acagcgacca
1981 cgctcttgaa gccctgtgcc tccagggact tcagcagatg ggtgtagagc gtggagccca
2041 gtcccgtccg ctggtggcgg gggctgacgt acacggtcga ctcggccgtc cagtcgtagg
2101 cgttgcgtgc cttccagggg cccgcgtagg cgatgccggc gacctcgccg tccacctcgg
2161 cgacgagcca gggatagcgc tcccgcagac ggacgaggtc gtccgtccac tcctgcggtt
2221 cctgcggctc ggtacggaag ttgaccgtgc ttgtctcgat gtagtggttg acgatggtgc
2281 agaccgccgg catgtccgcc tcggtggcac ggcggatgtc ggccgggcgt cgttctgggc
2341 tcatggtaga ctcgagagag atagatttgt agagagagac tggtgatttc agcgtgtcct
2401 ctccaaatga aatgaacttc cttatataga ggaagggtct tgcgaaggat agtgggattg
2461 tgcgtcatcc cttacgtcag tggagatatc acatcaatcc acttgctttg aagacgtggt
2521 tggaacgtct tctttttcca cgatgctcct cgtgggtggg ggtccatctt tgggaccact
2581 gtcggcagag gcatcttgaa cgatagcctt tcctttatcg caatgatggc atttgtaggt
2641 gccaccttcc ttttctactg tccttttgat gaagtgacag atagctgggc aatggaatcc
2701 gaggaggttt cccgatatta ccctttgttg aaaagtctca atagcccttt ggtcttctga
2761 gactgtatct ttgatattct tggagtagac gagagtgtcg tgctccacca tgttcacatc
2821 aatccacttg ctttgaagac gtggttggaa cgtcttcttt ttccacgatg ctcctcgtgg
2881 gtgggggtcc atctttggga ccactgtcgg cagaggcatc ttgaacgata gcctttcctt
2941 tatcgcaatg atggcatttg taggtgccac cttccttttc tactgtcctt ttgatgaagt
3001 gacagatagc tgggcaatgg aatccgagga ggtttcccga tattaccctt tgttgaaaag
3061 tctcaatagc cctttggtct tctgagactg tatctttgat attcttggag tagacgagag
3121 tgtcgtgctc caccatgttg gcaagctgct ctagccaata cgcaaaccgc ctctccccgc
3181 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
3241 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
3301 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
3361 cagctatgac atgattacga attcgagctc ggtacccggg gatcggcgcg ccagatttgc
3421 cttttcaatt tcagaaagaa tgctaaccca cagatggtta gagaggctta cgcagcaggt
3481 atcatcaaga cgatctaccc gagcaataat ctccaggaaa tcaaatacct tcccaagaag
3541 gttaaagatg cagtcaaaag attcaggact aactgcatca agaacacaga gaaagatata
3601 tttctcaaga tcagaagtac tattccagta tggacgattc aaggcttgct tcacaaacca
3661 aggcaagtaa tagagattgg agtctctaaa aaggtagttc ccactgaatc aaaggccatg
3721 gagtcaaaga ttcaaataga ggacctaaca gaactcgccg taaagactgg cgaacagttc
3781 atacagagtc tcttacgact caatgacaag aagaaaatct tcgtcaacat ggtggagcac
3841 gacacacttg tctactccaa aaatatcaaa gatacagtct cagaagacca aagggcaatt
3901 gagacttttc aacaaagggt aatatccgga aacctcctcg gattccattg cccagctatc
3961 tgtcacttta ttgtgaagat agtggaaaag gaaggtggct cctacaaatg ccatcattgc
4021 gataaaggaa aggccatcgt tgaagatgcc tctgccgaca gtggtcccaa agatggaccc
4081 ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg
4141 gattgatgtg atatctccac tgacgtaagg gatgacgcac aatcccacta tccttcgcaa
4201 gacccttcct ctatataagg aagttcattt catttggaga gaacacgggg gactcctgca
4261 ggaaaaatgg atcattatct tgatattaga cttagacctg atccagaatt tccaccagct
4321 caacttatgt ctgttctttt tggaaaactt catcaagctc ttgttgctca aggaggagat
4381 agaattggag tttcttttcc tgatcttgat gaatcaagat caagacttgg agaaagactt
4441 agaattcatg cttctgctga tgatcttaga gctttgcttg ctagaccttg gcttgaagga
4501 cttagagatc atcttcaatt tggagaacca gctgttgttc cacatccaac tccttataga
4561 caagtttcaa gagttcaagc taaatctaat ccagaaagac ttagaagaag acttatgaga
4621 agacatgatc tttctgaaga agaagctaga aaaagaattc ctgatactgt tgctagagct
4681 ttggatttgc cttttgttac acttagatca caatctactg gacaacattt tagacttttt
4741 attagacatg gaccacttca agttactgct gaagaaggag gatttacttg ttatggactt
4801 tctaagggag gttttgttcc ttggtttgga tctggagcta ctaatttttc tcttcttaag
4861 caagctggag atgttgaaga aaatcctgga cccatggata agaagtactc tatcggactc
4921 gatatcggaa ctaactctgt gggatgggct gtgatcaccg atgagtacaa ggtgccatct
4981 aagaagttca aggttctcgg aaacaccgat aggcactcta tcaagaaaaa ccttatcggt
5041 gctctcctct tcgattctgg tgaaactgct gaggctacca gactcaagag aaccgctaga
5101 agaaggtaca ccagaagaaa gaacaggatc tgctacctcc aagagatctt ctctaacgag
5161 atggctaaag tggatgattc attcttccac aggctcgaag agtcattcct cgtggaagaa
5221 gataagaagc acgagaggca ccctatcttc ggaaacatcg ttgatgaggt ggcataccac
5281 gagaagtacc ctactatcta ccacctcaga aagaagctcg ttgattctac tgataaggct
5341 gatctcaggc tcatctacct cgctctcgct cacatgatca agttcagagg acacttcctc
5401 atcgagggtg atctcaaccc tgataactct gatgtggata agttgttcat ccagctcgtg
5461 cagacctaca accagctttt cgaagagaac cctatcaacg cttcaggtgt ggatgctaag
5521 gctatcctct ctgctaggct ctctaagtca agaaggcttg agaacctcat tgctcagctc
5581 cctggtgaga agaagaacgg acttttcgga aacttgatcg ctctctctct cggactcacc
5641 cctaacttca agtctaactt cgatctcgct gaggatgcaa agctccagct ctcaaaggat
5701 acctacgatg atgatctcga taacctcctc gctcagatcg gagatcagta cgctgatttg
5761 ttcctcgctg ctaagaacct ctctgatgct atcctcctca gtgatatcct cagagtgaac
5821 accgagatca ccaaggctcc actctcagct tctatgatca agagatacga tgagcaccac
5881 caggatctca cacttctcaa ggctcttgtt agacagcagc tcccagagaa gtacaaagag
5941 attttcttcg atcagtctaa gaacggatac gctggttaca tcgatggtgg tgcatctcaa
6001 gaagagttct acaagttcat caagcctatc ctcgagaaga tggatggaac cgaggaactc
6061 ctcgtgaagc tcaatagaga ggatcttctc agaaagcaga ggaccttcga taacggatct
6121 atccctcatc agatccacct cggagagttg cacgctatcc ttagaaggca agaggatttc
6181 tacccattcc tcaaggataa cagggaaaag attgagaaga ttctcacctt cagaatccct
6241 tactacgtgg gacctctcgc tagaggaaac tcaagattcg cttggatgac cagaaagtct
6301 gaggaaacca tcaccccttg gaacttcgaa gaggtggtgg ataagggtgc tagtgctcag
6361 tctttcatcg agaggatgac caacttcgat aagaaccttc caaacgagaa ggtgctccct
6421 aagcactctt tgctctacga gtacttcacc gtgtacaacg agttgaccaa ggttaagtac
6481 gtgaccgagg gaatgaggaa gcctgctttt ttgtcaggtg agcaaaagaa ggctatcgtt
6541 gatctcttgt tcaagaccaa cagaaaggtg accgtgaagc agctcaaaga ggattacttc
6601 aagaaaatcg agtgcttcga ttcagttgag atttctggtg ttgaggatag gttcaacgca
6661 tctctcggaa cctaccacga tctcctcaag atcattaagg ataaggattt cttggataac
6721 gaggaaaacg aggatatctt ggaggatatc gttcttaccc tcaccctctt tgaagataga
6781 gagatgattg aagaaaggct caagacctac gctcatctct tcgatgataa ggtgatgaag
6841 cagttgaaga gaagaagata cactggttgg ggaaggctct caagaaagct cattaacgga
6901 atcagggata agcagtctgg aaagacaatc cttgatttcc tcaagtctga tggattcgct
6961 aacagaaact tcatgcagct catccacgat gattctctca cctttaaaga ggatatccag
7021 aaggctcagg tttcaggaca gggtgatagt ctccatgagc atatcgctaa cctcgctgga
7081 tctcctgcaa tcaagaaggg aatcctccag actgtgaagg ttgtggatga gttggtgaag
7141 gtgatgggaa ggcataagcc tgagaacatc gtgatcgaaa tggctagaga gaaccagacc
7201 actcagaagg gacagaagaa ctctagggaa aggatgaaga ggatcgagga aggtatcaaa
7261 gagcttggat ctcagatcct caaagagcac cctgttgaga acactcagct ccagaatgag
7321 aagctctacc tctactacct ccagaacgga agggatatgt atgtggatca agagttggat
7381 atcaacaggc tctctgatta cgatgttgat catatcgtgc cacagtcatt cttgaaggat
7441 gattctatcg ataacaaggt gctcaccagg tctgataaga acaggggtaa gagtgataac
7501 gtgccaagtg aagaggttgt gaagaaaatg aagaactatt ggaggcagct cctcaacgct
7561 aagctcatca ctcagagaaa gttcgataac ttgactaagg ctgagagggg aggactctct
7621 gaattggata aggcaggatt catcaagagg cagcttgtgg aaaccaggca gatcactaag
7681 cacgttgcac agatcctcga ttctaggatg aacaccaagt acgatgagaa cgataagttg
7741 atcagggaag tgaaggttat caccctcaag tcaaagctcg tgtctgattt cagaaaggat
7801 ttccaattct acaaggtgag ggaaatcaac aactaccacc acgctcacga tgcttacctt
7861 aacgctgttg ttggaaccgc tctcatcaag aagtatccta agctcgagtc agagttcgtg
7921 tacggtgatt acaaggtgta cgatgtgagg aagatgatcg ctaagtctga gcaagagatc
7981 ggaaaggcta ccgctaagta tttcttctac tctaacatca tgaatttctt caagaccgag
8041 attaccctcg ctaacggtga gatcagaaag aggccactca tcgagacaaa cggtgaaaca
8101 ggtgagatcg tgtgggataa gggaagggat ttcgctaccg ttagaaaggt gctctctatg
8161 ccacaggtga acatcgttaa gaaaaccgag gtgcagaccg gtggattctc taaagagtct
8221 atcctcccta agaggaactc tgataagctc attgctagga agaaggattg ggaccctaag
8281 aaatacggtg gtttcgattc tcctaccgtg gcttactctg ttctcgttgt ggctaaggtt
8341 gagaagggaa agagtaagaa gctcaagtct gttaaggaac ttctcggaat cactatcatg
8401 gaaaggtcat ctttcgagaa gaacccaatc gatttcctcg aggctaaggg atacaaagag
8461 gttaagaagg atctcatcat caagctccca aagtactcac tcttcgaact cgagaacggt
8521 agaaagagga tgctcgcttc tgctggtgag cttcaaaagg gaaacgagct tgctctccca
8581 tctaagtacg ttaactttct ttacctcgct tctcactacg agaagttgaa gggatctcca
8641 gaagataacg agcagaagca acttttcgtt gagcagcaca agcactactt ggatgagatc
8701 atcgagcaga tctctgagtt ctctaaaagg gtgatcctcg ctgatgcaaa cctcgataag
8761 gtgttgtctg cttacaacaa gcacagagat aagcctatca gggaacaggc agagaacatc
8821 atccatctct tcacccttac caacctcggt gctcctgctg ctttcaagta cttcgataca
8881 accatcgata ggaagagata cacctctacc aaagaagtgc tcgatgctac cctcatccat
8941 cagtctatca ctggactcta cgagactagg atcgatctct cacagctcgg tggtgattca
9001 agggctgatc ctaagaagaa gaggaaggtt tgactcgaga tatgaagatg aagatgaaat
9061 atttggtgtg tcaaataaaa agcttgtgtg cttaagtttg tgtttttttc ttggcttgtt
9121 gtgttatgaa tttgtggctt tttctaatat taaatgaatg taagatcaca ttataatgaa
9181 taaacaaatg tttctataat ccattgtgaa tgttttgttg gatctcttct gcagcatata
9241 actactgtat gtgctatggt atggactatg gaatatgatt aaagataagg agctccggtg
9301 acggacggcg cgctggcaga catactgtcc cacaaatgaa gatggaatct gtaaaagaaa
9361 acgcgtgaaa taatgcgtct gacaaaggtt aggtcggctg cctttaatca ataccaaagt
9421 ggtccctacc acgatggaaa aactgtgcag tcggtttggc tttttctgac gaacaaataa
9481 gattcgtggc cgacaggtgg gggtccacca tgtgaaggca tcttcagact ccaataatgg
9541 agcaatgacg taagggctta cgaaataagt aagggtagtt tgggaaatgt ccactcaccc
9601 gtcagtctat aaatacttag cccctccctc attgttaagg gagcaaaatc tcagagagat
9661 agtcctagag agagaaagag agcaagtagc ctagaagtag tcaaggcggc gaagtattca
9721 ggcacgtggc caggaagaag aaaagccaag acgacgaaaa caggtaagag ctaagcttcc
9781 tgcaggttca ctgccgtata ggcagactat tctttggatg caaatgtttt agagctagaa
9841 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
9901 cgttcactgc cgtataggca ggttaatatt aggtgtcgta ggttttagag ctagaaatag
9961 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcgtt
10021 cactgccgta taggcagact attatttgga tgcaaatgtt ttagagctag aaatagcaag
10081 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcgttcact
10141 gccgtatagg caggaatttg tattctaaga tgagttttag agctagaaat agcaagttaa
10201 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgcg ttcactgccg
10261 tataggcaga ttaaaaatat tctgataagg ttttagagct agaaatagca agttaaaata
10321 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcgttca ctgccgtata
10381 ggcagctaaa gacataaatt tgaaagtttt agagctagaa atagcaagtt aaaataaggc
10441 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg cgttcactgc cgtataggca
10501 ggtcgatcga caagctcgag tttctccata ataatgtgtg agtagttccc agataaggga
10561 attagggttc ctatagggtt tcgctcatgt gttgagcata taagaaaccc ttagtatgta
10621 tttgtatttg taaaatactt ctatcaataa aatttctaat tcctaaaacc aaaatccagt
10681 actaaaatcc agatcccccg aattaccggt ggcactggcc gtcgttttac aacgtcgtga
10741 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
10801 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
10861 tggcgaatgc tagagcagct tgagcttgga tcagattgtc gtttcccgcc ttcagtttaa
10921 actatcagtg tttgacagga tatattggcg ggtaaaccta agagaaaaga gcgtttatta
10981 gaataatcgg atatttaaaa gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtgc
11041 atgccaacca cagggttccc ctcgggatca aagtactttg atccaacccc tccgctgcta
11101 tagtgcagtc ggcttctgac gttcagtgca gccgtcttct gaaaacgaca tgtcgcacaa
11161 gtcctaagtt acgcgacagg ctgccgccct gcccttttcc tggcgttttc ttgtcgcgtg
11221 ttttagtcgc ataaagtaga atacttgcga ctagaaccgg agacattacg ccatgaacaa
11281 gagcgccgcc gctggcctgc tgggctatgc ccgcgtcagc accgacgacc aggacttgac
11341 caaccaacgg gccgaactgc acgcggccgg ctgcaccaag ctgttttccg agaagatcac
11401 cggcaccagg cgcgaccgcc cggagctggc caggatgctt gaccacctac gccctggcga
11461 cgttgtgaca gtgaccaggc tagaccgcct ggcccgcagc acccgcgacc tactggacat
11521 tgccgagcgc atccaggagg ccggcgcggg cctgcgtagc ctggcagagc cgtgggccga
11581 caccaccacg ccggccggcc gcatggtgtt gaccgtgttc gccggcattg ccgagttcga
11641 gcgttcccta atcatcgacc gcacccggag cgggcgcgag gccgccaagg cccgaggcgt
11701 gaagtttggc ccccgcccta ccctcacccc ggcacagatc gcgcacgccc gcgagctgat
11761 cgaccaggaa ggccgcaccg tgaaagaggc ggctgcactg cttggcgtgc atcgctcgac
11821 cctgtaccgc gcacttgagc gcagcgagga agtgacgccc accgaggcca ggcggcgcgg
11881 tgccttccgt gaggacgcat tgaccgaggc cgacgccctg gcggccgccg agaatgaacg
11941 ccaagaggaa caagcatgaa accgcaccag gacggccagg acgaaccgtt tttcattacc
12001 gaagagatcg aggcggagat gatcgcggcc gggtacgtgt tcgagccgcc cgcgcacggc
12061 tcaaccgtgc ggctgcatga aatcctggcc ggtttgtctg atgccaagct ggcggcctgg
12121 ccggccagct tggccgctga agaaaccgag cgccgccgtc taaaaaggtg atgtgtattt
12181 gagtaaaaca gcttgcgtca tgcggtcgct gcgtatatga tgcgatgagt aaataaacaa
12241 atacgcaagg ggaacgcatg aaggttatcg ctgtacttaa ccagaaaggc gggtcaggca
12301 agacgaccat cgcaacccat ctagcccgcg ccctgcaact cgccggggcc gatgttctgt
12361 tagtcgattc cgatccccag ggcagtgccc gcgattgggc ggccgtgcgg gaagatcaac
12421 cgctaaccgt tgtcggcatc gaccgcccga cgattgaccg cgacgtgaag gccatcggcc
12481 ggcgcgactt cgtagtgatc gacggagcgc cccaggcggc ggacttggct gtgtccgcga
12541 tcaaggcagc cgacttcgtg ctgattccgg tgcagccaag cccttacgac atatgggcca
12601 ccgccgacct ggtggagctg gttaagcagc gcattgaggt cacggatgga aggctacaag
12661 cggcctttgt cgtgtcgcgg gcgatcaaag gcacgcgcat cggcggtgag gttgccgagg
12721 cgctggccgg gtacgagctg cccattcttg agtcccgtat cacgcagcgc gtgagctacc
12781 caggcactgc cgccgccggc acaaccgttc ttgaatcaga acccgagggc gacgctgccc
12841 gcgaggtcca ggcgctggcc gctgaaatta aatcaaaact catttgagtt aatgaggtaa
12901 agagaaaatg agcaaaagca caaacacgct aagtgccggc cgtccgagcg cacgcagcag
12961 caaggctgca acgttggcca gcctggcaga cacgccagcc atgaagcggg tcaactttca
13021 gttgccggcg gaggatcaca ccaagctgaa gatgtacgcg gtacgccaag gcaagaccat
13081 taccgagctg ctatctgaat acatcgcgca gctaccagag taaatgagca aatgaataaa
13141 tgagtagatg aattttagcg gctaaaggag gcggcatgga aaatcaagaa caaccaggca
13201 ccgacgccgt ggaatgcccc atgtgtggag gaacgggcgg ttggccaggc gtaagcggct
13261 gggttgcctg ccggccctgc aatggcactg gaacccccaa gcccgaggaa tcggcgtgag
13321 cggtcgcaaa ccatccggcc cggtacaaat cggcgcggcg ctgggtgatg acctggtgga
13381 gaagttgaag gccgcgcagg ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg
13441 tgaatcgtgg caagcggccg ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc
13501 cggtgcgccg tcgattagga agccgcccaa gggcgacgag caaccagatt ttttcgttcc
13561 gatgctctat gacgtgggca cccgcgatag tcgcagcatc atggacgtgg ccgttttccg
13621 tctgtcgaag cgtgaccgac gagctggcga ggtgatccgc tacgagcttc cagacgggca
13681 cgtagaggtt tccgcagggc cggccggcat ggccagtgtg tgggattacg acctggtact
13741 gatggcggtt tcccatctaa ccgaatccat gaaccgatac cgggaaggga agggagacaa
13801 gcccggccgc gtgttccgtc cacacgttgc ggacgtactc aagttctgcc ggcgagccga
13861 tggcggaaag cagaaagacg acctggtaga aacctgcatt cggttaaaca ccacgcacgt
13921 tgccatgcag cgtacgaaga aggccaagaa cggccgcctg gtgacggtat ccgagggtga
13981 agccttgatt agccgctaca agatcgtaaa gagcgaaacc gggcggccgg agtacatcga
14041 gatcgagcta gctgattgga tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct
14101 gacggttcac cccgattact ttttgatcga tcccggcatc ggccgttttc tctaccgcct
14161 ggcacgccgc gccgcaggca aggcagaagc cagatggttg ttcaagacga tctacgaacg
14221 cagtggcagc gccggagagt tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc
14281 aaatgacctg ccggagtacg atttgaagga ggaggcgggg caggctggcc cgatcctagt
14341 catgcgctac cgcaacctga tcgagggcga agcatccgcc ggttcctaat gtacggagca
14401 gatgctaggg caaattgccc tagcagggga aaaaggtcga aaaggcctct ttcctgtgga
14461 tagcacgtac attgggaacc caaagccgta cattgggaac cggaacccgt acattgggaa
14521 cccaaagccg tacattggga accggtcaca catgtaagtg actgatataa aagagaaaaa
14581 aggcgatttt tccgcctaaa actctttaaa acttattaaa actcttaaaa cccgcctggc
14641 ctgtgcataa ctgtctggcc agcgcacagc ccaagagctg caaaaagcgc ctacccttcg
14701 gtcgctgcgc tccctacgcc ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc
14761 aaaaatggct ggcctacggc caggcaatct accagggcgc ggacaagccg cgccgtcgcc
14821 actcgaccgc cggcgcccac atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg
14881 aaaacctctg acacatgcag ctcccggaaa cggtcacagc ttgtctgtaa gcggatgccg
14941 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca
15001 tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg catcagagca
15061 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
15121 ataccgcatc aggccctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
15181 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
15241 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
15301 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
15361 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
15421 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
15481 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
15541 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
15601 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
15661 actggcagca gccactggta acaggattag cagagcgagg tat
//
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