| 质粒名称: | pHSN501 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 50589 |
| 存储实验室: | Qi-Jun Chen |
|
载体骨架基本信息 |
|
| 载体名称: | pGreen-like binary vector |
| 载体出品公司: | Mullineaux Lab, see http://www.pgreen.ac.uk/ |
| 载体抗性: | 卡那霉素和大观霉素 |
| 载体类型: | CRISPR ; 植物表达 |
| 筛选标记: | 潮霉素 |
| 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: | zCas9D10A,Cas9 |
| 物种来源: | Synthetic |
| 基因突变: | Cas9D10A nickase, was derived from the zCas9 and included in the toolkit to facilitate HR-based gene replacement or NHEJ-based insertion/deletion events (indels) through offset nicking under the precondition of the avoidance of off-target effects |
| 启动子 | 2×35Sp |
| 标签/融合蛋白: |
3xFLAG（插入件上的N端子）
NLS（插入件上的N端子）
|
| 5' 测序引物 | hSpCas9-R1, CGCTCGTGCTTCTTATCCTC |
|
文献引用方法 |
|
| 材料和方法部分: | pHSN501 was a gift from Qi-Jun Chen (Addgene plasmid # 50589 ; http://n2t.net/addgene:50589 ; RRID:Addgene_50589) |
| 参考文献部分: | A CRISPR/Cas9 toolkit for multiplex genome editing in plants. Xing HL, Dong L, Wang ZP, Zhang HY, Han CY, Liu B, Wang XC, Chen QJ. BMC Plant Biol. 2014 Nov 29;14(1):327. 10.1186/s12870-014-0327-y PubMed 25432517 |
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 pHSN501 12729 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION CRISPR/Cas based plant genome editing and gene regulation; expresses
zCas9D10A, gRNA scaffold for insertion of target sequence (AtU6-26
promoter), Hyg resistance.
ACCESSION .
VERSION .
KEYWORDS pHSN501
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12729)
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 12729)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..12729
/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"
CDS 810..1601
/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 1753..1828
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 2442..2786
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
primer_bind 2730..2750
/label=35S promoter
/note="CaMV 35S promoter, forward primer"
CDS 2969..3034
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 3041..3061
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 3086..7186
/codon_start=1
/product="nickase mutant of the Cas9 endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system"
/label=Cas9(D10A)
/note="generates RNA-guided single strand nicks in DNA due
to the D10A mutation in the RuvC catalytic domain"
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
RBS 7099..7107
/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 7187..7234
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
terminator 7285..7537
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
promoter 7653..8330
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
CDS 8398..9423
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
K"
polyA_signal 9463..9637
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
misc_feature 9787..9811
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
primer_bind complement(10059..10082)
/label=pENTR-F
/note="pENTR vectors, forward primer"
terminator 10090..10117
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 10209..10295
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
rep_origin 10464..10899
/label=pSa ori
/note="origin of replication from bacterial plasmid pSa"
rep_origin complement(11014..11602)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(11094..11113)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(11692..12501)
/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 12415..12434
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 12493..12512
/label=pENTR-R
/note="pENTR vectors, reverse primer"
ORIGIN
1 gtttacccgc caatatatcc tgtcaaacac tgatagttta aactgaaggc gggaaacgac
61 aatctgatcc aagctcaagc taagctcacg tgacggaatt aagcttcgac ttgccttccg
121 cacaatacat catttcttct tagctttttt tcttcttctt cgttcataca gttttttttt
181 gtttatcagc ttacattttc ttgaaccgta gctttcgttt tcttcttttt aactttccat
241 tcggagtttt tgtatcttgt ttcatagttt gtcccaggat tagaatgatt aggcatcgaa
301 ccttcaagaa tttgattgaa taaaacatct tcattcttaa gatatgaaga taatcttcaa
361 aaggcccctg ggaatctgaa agaagagaag caggcccatt tatatgggaa agaacaatag
421 tatttcttat ataggcccat ttaagttgaa aacaatcttc aaaagtccca catcgcttag
481 ataagaaaac gaagctgagt ttatatacag ctagagtcga agtagtgatt gggagaccaa
541 cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt atgcgctcac
601 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca
661 tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag
721 cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg
781 gcagtcgccc taaaacaaag ttaaacatca tgggggaagc ggtgatcgcc gaagtatcga
841 ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg
901 tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc
961 tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt
1021 tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg
1081 ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag
1141 aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc
1201 tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg
1261 aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa
1321 cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt
1381 cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact
1441 gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt
1501 atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc
1561 actacgtgaa aggcgagatc accaaggtag tcggcaaata atgtctagct agaaattcgt
1621 tcaagccgac gccgcttcgc ggcgcggctt aactcaagcg ttagatgcac taagcacata
1681 attgctcaca gccaaactat caggtcaagt ctgcttttat tatttttaag cgtgcataat
1741 aagccggtct cggttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca
1801 acttgaaaaa gtggcaccga gtcggtgctt ttttttgcaa aattttccag atcgatttct
1861 tcttcctctg ttcttcggcg ttcaatttct ggggttttct cttcgttttc tgtaactgaa
1921 acctaaaatt tgacctaaaa aaaatctcaa ataatatgat tcagtggttt tgtacttttc
1981 agttagttga gttttgcagt tccgatgaga taaaccaata agcttgcatg cctgcaggtc
2041 aacatggtgg agcacgacac acttgtctac tccaaaaata tcaaagatac agtctcagaa
2101 gaccaaaggg caattgagac ttttcaacaa agggtaatat ccggaaacct cctcggattc
2161 cattgcccag ctatctgtca ctttattgtg aagatagtgg aaaaggaagg tggctcctac
2221 aaatgccatc attgcgataa aggaaaggcc atcgttgaag atgcctctgc cgacagtggt
2281 cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagacgt tccaaccacg
2341 tcttcaaagc aagtggattg atgtgataac atggtggagc acgacacact tgtctactcc
2401 aaaaatatca aagatacagt ctcagaagac caaagggcaa ttgagacttt tcaacaaagg
2461 gtaatatccg gaaacctcct cggattccat tgcccagcta tctgtcactt tattgtgaag
2521 atagtggaaa aggaaggtgg ctcctacaaa tgccatcatt gcgataaagg aaaggccatc
2581 gttgaagatg cctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc
2641 gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag tggattgatg tgatatctcc
2701 actgacgtaa gggatgacgc acaatcccac tatccttcgc aagacccttc ctctatataa
2761 ggaagttcat ttcatttgga gaggacctcg acctcaacac aacatataca aaacaaacga
2821 atctcaagca atcaagcatt ctacttctat tgcagcaatt taaatcattt cttttaaagc
2881 aaaagcaatt ttctgaaaat tttcaccatt tacgaacgat actcgagggg gatccccaat
2941 acttgtatgg ccgcggccgc tctagatgga ttacaaggac cacgacgggg attacaagga
3001 ccacgacatt gattacaagg atgatgatga caagatggct ccgaagaaga agaggaaggt
3061 tggcatccac ggggtgccag ctgctgacaa gaagtactcg atcggcctcg ccattgggac
3121 taactctgtt ggctgggccg tgatcaccga cgagtacaag gtgccctcaa agaagttcaa
3181 ggtcctgggc aacaccgatc ggcattccat caagaagaat ctcattggcg ctctcctgtt
3241 cgacagcggc gagacggctg aggctacgcg gctcaagcgc accgcccgca ggcggtacac
3301 gcgcaggaag aatcgcatct gctacctgca ggagattttc tccaacgaga tggcgaaggt
3361 tgacgattct ttcttccaca ggctggagga gtcattcctc gtggaggagg ataagaagca
3421 cgagcggcat ccaatcttcg gcaacattgt cgacgaggtt gcctaccacg agaagtaccc
3481 tacgatctac catctgcgga agaagctcgt ggactccaca gataaggcgg acctccgcct
3541 gatctacctc gctctggccc acatgattaa gttcaggggc catttcctga tcgaggggga
3601 tctcaacccg gacaatagcg atgttgacaa gctgttcatc cagctcgtgc agacgtacaa
3661 ccagctcttc gaggagaacc ccattaatgc gtcaggcgtc gacgcgaagg ctatcctgtc
3721 cgctaggctc tcgaagtctc ggcgcctcga gaacctgatc gcccagctgc cgggcgagaa
3781 gaagaacggc ctgttcggga atctcattgc gctcagcctg gggctcacgc ccaacttcaa
3841 gtcgaatttc gatctcgctg aggacgccaa gctgcagctc tccaaggaca catacgacga
3901 tgacctggat aacctcctgg cccagatcgg cgatcagtac gcggacctgt tcctcgctgc
3961 caagaatctg tcggacgcca tcctcctgtc tgatattctc agggtgaaca ccgagattac
4021 gaaggctccg ctctcagcct ccatgatcaa gcgctacgac gagcaccatc aggatctgac
4081 cctcctgaag gcgctggtca ggcagcagct ccccgagaag tacaaggaga tcttcttcga
4141 tcagtcgaag aacggctacg ctgggtacat tgacggcggg gcctctcagg aggagttcta
4201 caagttcatc aagccgattc tggagaagat ggacggcacg gaggagctgc tggtgaagct
4261 caatcgcgag gacctcctga ggaagcagcg gacattcgat aacggcagca tcccacacca
4321 gattcatctc ggggagctgc acgctatcct gaggaggcag gaggacttct accctttcct
4381 caaggataac cgcgagaaga tcgagaagat tctgactttc aggatcccgt actacgtcgg
4441 cccactcgct aggggcaact cccgcttcgc ttggatgacc cgcaagtcag aggagacgat
4501 cacgccgtgg aacttcgagg aggtggtcga caagggcgct agcgctcagt cgttcatcga
4561 gaggatgacg aatttcgaca agaacctgcc aaatgagaag gtgctcccta agcactcgct
4621 cctgtacgag tacttcacag tctacaacga gctgactaag gtgaagtatg tgaccgaggg
4681 catgaggaag ccggctttcc tgtctgggga gcagaagaag gccatcgtgg acctcctgtt
4741 caagaccaac cggaaggtca cggttaagca gctcaaggag gactacttca agaagattga
4801 gtgcttcgat tcggtcgaga tctctggcgt tgaggaccgc ttcaacgcct ccctggggac
4861 ctaccacgat ctcctgaaga tcattaagga taaggacttc ctggacaacg aggagaatga
4921 ggatatcctc gaggacattg tgctgacact cactctgttc gaggaccggg agatgatcga
4981 ggagcgcctg aagacttacg cccatctctt cgatgacaag gtcatgaagc agctcaagag
5041 gaggaggtac accggctggg ggaggctgag caggaagctc atcaacggca ttcgggacaa
5101 gcagtccggg aagacgatcc tcgacttcct gaagagcgat ggcttcgcga accgcaattt
5161 catgcagctg attcacgatg acagcctcac attcaaggag gatatccaga aggctcaggt
5221 gagcggccag ggggactcgc tgcacgagca tatcgcgaac ctcgctggct cgccagctat
5281 caagaagggg attctgcaga ccgtgaaggt tgtggacgag ctggtgaagg tcatgggcag
5341 gcacaagcct gagaacatcg tcattgagat ggcccgggag aatcagacca cgcagaaggg
5401 ccagaagaac tcacgcgaga ggatgaagag gatcgaggag ggcattaagg agctggggtc
5461 ccagatcctc aaggagcacc cggtggagaa cacgcagctg cagaatgaga agctctacct
5521 gtactacctc cagaatggcc gcgatatgta tgtggaccag gagctggata ttaacaggct
5581 cagcgattac gacgtcgatc atatcgttcc acagtcattc ctgaaggatg actccattga
5641 caacaaggtc ctcaccaggt cggacaagaa ccggggcaag tctgataatg ttccttcaga
5701 ggaggtcgtt aagaagatga agaactactg gcgccagctc ctgaatgcca agctgatcac
5761 gcagcggaag ttcgataacc tcacaaaggc tgagaggggc gggctctctg agctggacaa
5821 ggcgggcttc atcaagaggc agctggtcga gacacggcag atcactaagc acgttgcgca
5881 gattctcgac tcacggatga acactaagta cgatgagaat gacaagctga tccgcgaggt
5941 gaaggtcatc accctgaagt caaagctcgt ctccgacttc aggaaggatt tccagttcta
6001 caaggttcgg gagatcaaca attaccacca tgcccatgac gcgtacctga acgcggtggt
6061 cggcacagct ctgatcaaga agtacccaaa gctcgagagc gagttcgtgt acggggacta
6121 caaggtttac gatgtgagga agatgatcgc caagtcggag caggagattg gcaaggctac
6181 cgccaagtac ttcttctact ctaacattat gaatttcttc aagacagaga tcactctggc
6241 caatggcgag atccggaagc gccccctcat cgagacgaac ggcgagacgg gggagatcgt
6301 gtgggacaag ggcagggatt tcgcgaccgt caggaaggtt ctctccatgc cacaagtgaa
6361 tatcgtcaag aagacagagg tccagactgg cgggttctct aaggagtcaa ttctgcctaa
6421 gcggaacagc gacaagctca tcgcccgcaa gaaggactgg gatccgaaga agtacggcgg
6481 gttcgacagc cccactgtgg cctactcggt cctggttgtg gcgaaggttg agaagggcaa
6541 gtccaagaag ctcaagagcg tgaaggagct gctggggatc acgattatgg agcgctccag
6601 cttcgagaag aacccgatcg atttcctgga ggcgaagggc tacaaggagg tgaagaagga
6661 cctgatcatt aagctcccca agtactcact cttcgagctg gagaacggca ggaagcggat
6721 gctggcttcc gctggcgagc tgcagaaggg gaacgagctg gctctgccgt ccaagtatgt
6781 gaacttcctc tacctggcct cccactacga gaagctcaag ggcagccccg aggacaacga
6841 gcagaagcag ctgttcgtcg agcagcacaa gcattacctc gacgagatca ttgagcagat
6901 ttccgagttc tccaagcgcg tgatcctggc cgacgcgaat ctggataagg tcctctccgc
6961 gtacaacaag caccgcgaca agccaatcag ggagcaggct gagaatatca ttcatctctt
7021 caccctgacg aacctcggcg cccctgctgc tttcaagtac ttcgacacaa ctatcgatcg
7081 caagaggtac acaagcacta aggaggtcct ggacgcgacc ctcatccacc agtcgattac
7141 cggcctctac gagacgcgca tcgacctgtc tcagctcggg ggcgacaagc ggccagcggc
7201 gacgaagaag gcggggcagg cgaagaagaa gaagtgagct caggcctcca tacaagtatt
7261 ggggatccac tagcgaattt ccccgatcgt tcaaacattt ggcaataaag tttcttaaga
7321 ttgaatcctg ttgccggtct tgcgatgatt atcatataat ttctgttgaa ttacgttaag
7381 catgtaataa ttaacatgta atgcatgacg ttatttatga gatgggtttt tatgattaga
7441 gtcccgcaat tatacattta atacgcgata gaaaacaaaa tatagcgcgc aaactaggat
7501 aaattatcgc gcgcggtgtc atctatgtta ctagatcgga attcgaggcg gtttgcgtat
7561 tggctagagc agcttgccaa catggtggag cacgacactc tcgtctactc caagaatatc
7621 aaagatacag tctcagaaga ccaaagggct attgagactt ttcaacaaag ggtaatatcg
7681 ggaaacctcc tcggattcca ttgcccagct atctgtcact tcatcaaaag gacagtagaa
7741 aaggaaggtg gcacctacaa atgccatcat tgcgataaag gaaaggctat cgttcaagat
7801 gcctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat cgtggaaaaa
7861 gaagacgttc caaccacgtc ttcaaagcaa gtggattgat gtgataacat ggtggagcac
7921 gacactctcg tctactccaa gaatatcaaa gatacagtct cagaagacca aagggctatt
7981 gagacttttc aacaaagggt aatatcggga aacctcctcg gattccattg cccagctatc
8041 tgtcacttca tcaaaaggac agtagaaaag gaaggtggca cctacaaatg ccatcattgc
8101 gataaaggaa aggctatcgt tcaagatgcc tctgccgaca gtggtcccaa agatggaccc
8161 ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg
8221 gattgatgtg atatctccac tgacgtaagg gatgacgcac aatcccacta tccttcgcaa
8281 gaccttcctc tatataagga agttcatttc atttggagag gacacgctga aatcaccagt
8341 ctctctctac aaatctatct ctctcgagct ttcgcagatc ccggggggca atgagatatg
8401 aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc
8461 gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta
8521 ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt
8581 tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg
8641 gagtttagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa
8701 gacctgcctg aaaccgaact gcccgctgtt ctacaaccgg tcgcggaggc tatggatgcg
8761 atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc
8821 ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac
8881 tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg
8941 atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc
9001 aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg
9061 ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt
9121 atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccacga
9181 ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc
9241 aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc
9301 gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt
9361 gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaagaaa
9421 tagagtagat gccgaccgga tctgtcgatc gacaagctcg agtttctcca taataatgtg
9481 tgagtagttc ccagataagg gaattagggt tcctataggg tttcgctcat gtgttgagca
9541 tataagaaac ccttagtatg tatttgtatt tgtaaaatac ttctatcaat aaaatttcta
9601 attcctaaaa ccaaaatcca gtactaaaat ccagatcccc cgaattaatt cggcgttaat
9661 tcagtacatt aaaaacgtcc gcaatgtgtt attaagttgt cactagtcag gttaactcaa
9721 ttcggcgtta attcagtaca ttaaaaacgt ccgcaatgtg ttattaagtt gtctaagcgt
9781 caatttgttt acaccacaat atatcctgcc accagccagc caacagctcc ccgaccggca
9841 gctcggcaca aaatcaccac tcgatacagg cagcccatca gtccgggacg gcgtcagcgg
9901 gagagccgtt gtaaggcggc agactttgct catgttaccg atgctattcg gaagaacggc
9961 aactaagctg ccgggtttga aacacggatg atctcgcgga gggtagcatg ttgattgtaa
10021 cgatgacaga gcgttgctgc ctgtgatcac ttaagtaact aactaacagg aagagtttgt
10081 agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt
10141 tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct
10201 cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag
10261 gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc
10321 ttagtagtta gacgtccccg agatccatgc tagaccatga atccagaagc ccgagaggtt
10381 gccgcctttc gggctttttc tttttcaaaa aaaaaaattt ataaaacgat ctgttgcggc
10441 cggccgccgg gttgtgggca aaggcgctcg acggtgggca accgcttgcg gttgtccacg
10501 ggcggagccg gtgcgcgtag cgcattgtcc acaagccaag ggcgaccaat aattgatata
10561 tatattcata attgaaaagc taattgaaca tactacttgc tgtaactact tgccggagcg
10621 aggggtgttt gcaagctgtt gatctgaaag ggctattagc gttctcacgt gcctttttga
10681 ttagcgattt cacgtgacct tattagcgat ttcacgtact ccgattagcg atttcacgta
10741 ccctgattag cgatttcacg tggatagttt ttggagcggg ccggaaagcc ccgtgaatca
10801 aggctttgcg gggcattagc ggtttcacgt ggataactac cctctatcca caggcttccg
10861 gggataaaaa agcccgctcg acggcgggct gttggatggg gatctagcgg taatacggtt
10921 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
10981 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
11041 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
11101 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
11161 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
11221 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
11281 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
11341 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
11401 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
11461 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
11521 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
11581 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctaccgggt ctgacgctca
11641 gtggaacggg gcccaatctg aataatgtta caaccaatta accaattctg attagaaaaa
11701 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
11761 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
11821 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
11881 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
11941 tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
12001 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
12061 gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
12121 gcgcagggac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
12181 tacctggaat gctgtttttc cggggatcgc agtggtgagt aaccatgcat catcaggagt
12241 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
12301 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
12361 cgcatcgggc ttcccataca agcgatagat tgtcgcacct gattgcccga cattatcgcg
12421 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctcgacgt
12481 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
12541 tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac
12601 gggccagagc tgcagtttga tcccgagggg aaccctgtgg ttgacatgca catacaaatg
12661 gacgaacgga taaacctttt cacgcccttt taaatatccg ttattctaat aaacgctctt
12721 ttctcttag
//
