| 质粒名称: | pHSP02 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 117259 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pCas |
| 载体出品公司: | Sheng Yang, Addgene Plasmid #62225 |
| 载体抗性: | 卡那霉素低（25微克/毫升） |
| 空载体大小: | 4476 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | -- |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 30 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Cas9, promotor P11-guide-RNA, homologous arms of Lp_0537,Cas9 (4107 bps), promoter-P11-guide-RNA (144 bps), homologous arm (2000 bps) |
| 物种来源: | Cas9 (Streptococcus pyogenes M1 GAS) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | Cas9: AE004092 (SPy_1046) |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | XbaI(未破坏) |
| 3' 克隆位点: | ApaI(未破坏) |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pHSP02 was a gift from Sheng Yang (Addgene plasmid # 117259 ; http://n2t.net/addgene:117259 ; RRID:Addgene_117259) |
| 参考文献部分: | Development of a RecE/T-Assisted CRISPR-Cas9 Toolbox for Lactobacillus. Huang H, Song X, Yang S. Biotechnol J. 2019 Jul;14(7):e1800690. doi: 10.1002/biot.201800690. Epub 2019 May 20. 10.1002/biot.201800690 PubMed 30927506 |
买家导航
pHSP02载体质粒基本信息
pHSP02质粒图谱载体图谱和pHSP02载体序列质粒序列多克隆位点信息
pHSP02质粒载体简介
pHSP02质粒序列载体序列
LOCUS pHSP02 14102 bp ds-DNA circular SYN 17-JUN-2019
DEFINITION Genome editing for Lactobacillus plantarum WCSF1.
ACCESSION .
VERSION .
KEYWORDS pHSP02
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14102)
TITLE RecE/T-Assisted Genome Editing in Lactobacillus
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 14102)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jun 17, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..14102
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(319..1134)
/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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 1042..1061
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 1126..1145
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 1431..5537
/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"
misc_RNA complement(12299..12374)
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
CDS complement(12549..13499)
/codon_start=1
/gene="rep101 encoding an A56V mutation"
/product="temperature-sensitive version of the RepA protein
needed for replication with the pSC101 origin (Armstrong et
al., 1984)"
/label=Rep101(Ts)
/translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
rep_origin 13547..13769
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
ORIGIN
1 ggtctgctat gtggtgctat ctgacttttt gctgttcagc agttcctgcc ctctgatttt
61 ccagtctgac cacttcggat tatcccgtga caggtcattc agactggcta atgcacccag
121 taaggcagcg gtatcatcaa cggggtctga cgctcagtgg aacgaaaact cacgttaagg
181 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
241 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt acgtttccaa
301 ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
361 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
421 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
481 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
541 atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca tttctttcca
601 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
661 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
721 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
781 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg ggatcgcagt
841 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
901 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
961 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc gatagattgt
1021 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
1081 gttggaattt aatcgcggcc tcgagcaaga cgtttcccgt tgaatatggc tcataacacc
1141 ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc
1201 ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttcctgca gcgaaaagcc
1261 ctgacaaccc ttgttcctaa aaaggaataa gcgttcggtc agtaaataat agaaataaaa
1321 aatcagacct aagactgatg acaaaaagag aaaattttga taaaatagtc ttagaattaa
1381 attaaaaagg gaggccaaat ataatgaaaa atatgaatga caatgatgtt atggataaga
1441 aatactcaat aggcttagat atcggcacaa atagcgtcgg atgggcggtg atcactgatg
1501 aatataaggt tccgtctaaa aagttcaagg ttctgggaaa tacagaccgc cacagtatca
1561 aaaaaaatct tataggggct cttttatttg acagtggaga gacagcggaa gcgactcgtc
1621 tcaaacggac agctcgtaga aggtatacac gtcggaagaa tcgtatttgt tatctacagg
1681 agattttttc aaatgagatg gcgaaagtag atgatagttt ctttcatcga cttgaagagt
1741 cttttttggt ggaagaagac aagaagcatg aacgtcatcc tatttttgga aatatagtag
1801 atgaagttgc ttatcatgag aaatatccaa ctatctatca tctgcgaaaa aaattggtag
1861 attctactga taaagcggat ttgcgcttaa tctatttggc cttagcgcat atgattaagt
1921 ttcgtggtca ttttttgatt gagggagatt taaatcctga taatagtgat gtggacaaac
1981 tatttatcca gttggtacaa acctacaatc aattatttga agaaaaccct attaacgcaa
2041 gtggagtaga tgctaaagcg attctttctg cacgattgag taaatcaaga cgattagaaa
2101 atctcattgc tcagctcccc ggtgagaaga aaaatggctt atttgggaat ctcattgctt
2161 tgtcattggg tttgacccct aattttaaat caaattttga tttggcagaa gatgctaaat
2221 tacagctttc aaaagatact tacgatgatg atttagataa tttattggcg caaattggag
2281 atcaatatgc tgatttgttt ttggcagcta agaatttatc agatgctatt ttactttcag
2341 atatcctaag agtaaatact gaaataacta aggctcccct atcagcttca atgattaaac
2401 gctacgatga acatcatcaa gacttgactc ttttaaaagc tttagttcga caacaacttc
2461 cagaaaagta taaagaaatc ttttttgatc aatcaaaaaa cggatatgca ggttatattg
2521 atgggggagc tagccaagaa gaattttata aatttatcaa accaatttta gaaaaaatgg
2581 atggtactga ggaattattg gtgaaactaa atcgtgaaga tttgctgcgc aagcaacgga
2641 cctttgacaa cggctctatt ccccatcaaa ttcacttggg tgagctgcat gctattttga
2701 gaagacaaga agacttttat ccatttttaa aagacaatcg tgagaagatt gaaaaaatct
2761 tgacttttcg aattccttat tatgttggtc cattggcgcg tggcaatagt cgttttgcat
2821 ggatgactcg gaagtctgaa gaaacaatta ccccatggaa ttttgaagaa gttgtcgata
2881 aaggtgcttc agctcaatca tttattgaac gcatgacaaa ctttgataaa aatcttccaa
2941 atgaaaaagt actaccaaaa catagtttgc tttatgagta ttttacggtt tataacgaat
3001 tgacaaaggt caaatatgtt actgaaggaa tgcgaaaacc agcatttctt tcaggtgaac
3061 agaagaaagc cattgttgat ttactcttca aaacaaatcg aaaagtaacc gttaagcaat
3121 taaaagaaga ttatttcaaa aaaatagaat gttttgatag tgttgaaatt tcaggagttg
3181 aagatagatt taatgcttca ttaggtacct accatgattt gctaaaaatt attaaagata
3241 aagatttttt ggataatgaa gaaaatgaag atatcttaga ggatattgtt ttaacattga
3301 ccttatttga agatagggag atgattgagg aaagacttaa aacatatgct cacctctttg
3361 atgataaggt gatgaaacag cttaaacgtc gccgttatac tggttgggga cgtttgtctc
3421 gaaaattgat taatggtatt agggataagc aatctggcaa aacaatatta gattttttga
3481 aatcagatgg ttttgccaat cgcaatttta tgcagctgat ccatgatgat agtttgacat
3541 ttaaagaaga cattcaaaaa gcacaagtgt ctggacaagg cgatagttta catgaacata
3601 ttgcaaattt agctggtagc cctgctatta aaaaaggtat tttacagact gtaaaagttg
3661 ttgatgaatt ggtcaaagta atggggcggc ataagccaga aaatatcgtt attgaaatgg
3721 cacgtgaaaa tcagacaact caaaagggcc agaaaaattc gcgagagcgt atgaaacgaa
3781 tcgaagaagg tatcaaagaa ttaggaagtc agattcttaa agagcatcct gttgaaaata
3841 ctcaattgca aaatgaaaag ctctatctct attatctcca aaatggaaga gacatgtatg
3901 tggaccaaga attagatatt aatcgtttaa gtgattatga tgtcgatcac attgttccac
3961 aaagtttcct taaagacgat tcaatagaca ataaggtctt aacgcgttct gataaaaatc
4021 gtggtaaatc ggataacgtt ccaagtgaag aagtagtcaa aaagatgaaa aactattgga
4081 gacaacttct aaacgccaag ttaatcactc aacgtaagtt tgataattta acgaaagctg
4141 aacgtggagg tttgagtgaa cttgataaag ctggttttat caaacgccaa ttggttgaaa
4201 ctcgccaaat cactaagcat gtggcacaaa ttttggatag tcgcatgaat actaaatacg
4261 atgaaaatga taaacttatt cgagaggtta aagtgattac cttaaaatct aaattagttt
4321 ctgacttccg aaaagatttc caattctata aagtacgtga gattaacaat taccatcatg
4381 cccatgatgc gtatctaaat gccgtcgttg gaactgcttt gattaagaaa tatccaaaac
4441 ttgaatcgga gtttgtctat ggtgattata aagtttatga tgttcgtaaa atgattgcta
4501 agtctgagca agaaataggc aaagcaaccg caaaatattt cttttactct aatatcatga
4561 acttcttcaa aacagaaatt acacttgcaa atggagagat tcgcaaacgc cctctaatcg
4621 aaactaatgg ggaaactgga gaaattgtct gggataaagg gcgagatttt gccacagtgc
4681 gcaaagtatt gtccatgccc caagtcaata ttgtcaagaa aacagaagta cagacaggcg
4741 gattctccaa ggagtcaatt ttaccaaaaa gaaattcgga caagcttatt gctcgtaaaa
4801 aagactggga tccaaaaaaa tatggtggtt ttgatagtcc aacggtagct tattcagtcc
4861 tagtggttgc taaggtggaa aaagggaaat cgaagaagtt aaaatccgtt aaagagttac
4921 tagggatcac aattatggaa agaagttcct ttgaaaaaaa tccgattgac tttttagaag
4981 ctaaaggata taaggaagtt aaaaaagact taatcattaa actacctaaa tatagtcttt
5041 ttgagttaga aaacggtcgt aaacggatgc tggctagtgc cggagaatta caaaaaggaa
5101 atgagctggc tctgccaagc aaatatgtga attttttata tttagctagt cattatgaaa
5161 agttgaaggg tagtccagaa gataacgaac aaaaacaatt gtttgtggag cagcataagc
5221 attatttaga tgagattatt gagcaaatca gtgaattttc taagcgtgtt attttagcag
5281 atgccaattt agataaagtt cttagtgcat ataacaaaca tagagacaaa ccaatacgtg
5341 aacaagcaga aaatattatt catttattta cgttgacgaa tcttggagct cccgctgctt
5401 ttaaatattt tgatacaaca attgatcgta aacgatatac gtctacaaaa gaagttttag
5461 atgccactct tatccatcaa tccatcactg gtctttatga aacacgcatt gatttgagtc
5521 agctaggagg tgactgaagt atattttaga tgaagattat ttcttaatct aggctcgaga
5581 tctatcgata agcttaagtg attagtcaaa gaatggtgat gacaattgta aattctattt
5641 aatcactttg actagcaaat actaacaaca agacacacac accaaaaatc aaaaattcac
5701 tacttttagt taaaaaccac gtaaccacaa gaactaatcc aatccatgta atcgggttct
5761 tcaaatattt ctccaagatt ttcctcctct aatatgctca acttaaatga cctattcaat
5821 aaatctatta tgctgctaaa tagtttatag gacaaataag tatactctaa tgacctataa
5881 aagatagaaa attaaaaaat caagtgttcg cttcgctctc actgcccctc gacgttttag
5941 tagcctttcc ctcacttcgt tcagtccaag ccaactaaaa gttttcgggc tactctctcc
6001 ttctccccct aataattaat taaaatctta ctctgtatat ttctgctaat cattcgctaa
6061 acagcaaaga aaaaacaaac acgtatcata gatataaatg taatggcata gtgcgggttt
6121 tattttcagc ctgtatcata gctaaacaaa tcgagttgtg tgtccgtttt agggcgttct
6181 gctagcttgt ttaaagtctc ttgaatgaat gtatgctcta agtcaaaaga atttgtcagc
6241 gcctttatat agctttcttt ttcttctttt tttactttaa tgatcgatag caacaatgat
6301 ttaacactag caagttgaat gccaccattt cttcctggtt taatcttaaa gaaaatttcc
6361 tgattcgcct tcagtacctt cagcaattta tctaatgtcc gttcaggaat gcctagcact
6421 tctctaatct cttttttggt cgtcactaaa taaggcttgt atacatcgct tttttcgcta
6481 atataagcca ttaaatcttc tttccattct gacaaatgaa cacgttgacg ttcgcttctt
6541 tttttcttga atttaaacca cccttgacgg acaaataaat ctttactggt taaatcactt
6601 gatacccaag ctttgcaaag aatggtaatg tattccctat tagccccttg atagttttct
6661 gaataggcac ttctaacaat tttgattact tctttttctt ctaagggttg atctaatcga
6721 ttattaaact caaacatatt atattcgcac gtttcgattg aatagcctga actaaagtag
6781 gctaaagaga gggtaaacat gacgttatta cgccctatta aacccttttc tcctgaaaat
6841 ttcgtttcgt gcaataagag attaaaccag ggttcatcta cttgtttttt gccttctgta
6901 ccgcttaaaa ccgttagact tgaacgagta aagcccttat tatctgtttg tttgaaagac
6961 caatcttgcc attctttgaa agaataacgg taattaggat caaaaaattc tacattgtcc
7021 gttcttggta tgcgagcaat accaaaatga ttacacgtta gatcaactgg caaagacttt
7081 ccaaaatatt ctcggatatt ttgcgaaatt attttggctg ctttgacaga tttaaattct
7141 gattttgaag tcacatagac tggcgtttct aaaacaaaat atgcttgata acctttatca
7201 gatttgataa tcatagtagg cataaaacct aaatcaatag cggttgttaa aatatcgctt
7261 gctgaaatag tttcttttgc cgtgtgaata tcaaaatcaa taaagaaggt attgatttgt
7321 cttaaattgt tttcagaatg tcctttcgtg tatgaacggt tttcgtctgc atacgttcca
7381 taacgataaa cgttgggtgt ccaatgtgta aatgtatctt gattttcttg aatcgcttcc
7441 tcggaagtca gaacaacacc acgaccgcca atcatgcttg attttgagcg atacgcaaaa
7501 atagcccctt tgcttttacc tggcttggta gtgattgagc gaattttact atttttaaat
7561 ttgtacttta acaagccgtc atgaagcaca gtttctacaa caaaagggat attcattcag
7621 ctgttctcct ttcctataaa tcctataaaa taggttgttt aattaacttg gtttgctttt
7681 tcattcaact gtttcaatat tgcatgtttt gaaaaagatt tttttccttt ataagtcaat
7741 ttttttccac taatcgaata aattattttg ttattttcta ttaacttata tatataatct
7801 tccccctccg aagaaaaata cttatctgat tttgtttcta agtagatatt tctcttttct
7861 aactctttct taaacgtttc tagtgtatag atatttgcta attttcttat ctccaataaa
7921 ctatttttta tataagtttt acattcatca tgattcatac aaactccacc ttctataaat
7981 gaatacaaaa aaagcaatca aacgatttcc gattgattgc ttaacaattc ttaaattcag
8041 tagcttagat acttgaaaac tctctgattt ccctatataa tgatagtacg gttatatacc
8101 gtcttcaaac aaagttaatt aaataacttc ttacgaggga agagttcatc tgactaactg
8161 ataagcgttg gtttggcaat cttatcgggc tatgcattta taaaatgtcg tcaaacattt
8221 tataaatgtg tcatggctct tttttcgttt ctattcagtt cgttgtttcg ttatatctag
8281 tataccgctt ttaaaaaaaa taagcaacga tttcgtgcat tattcacacg aagtcattgc
8341 ttttttcttc ttccatttct aaatccaatg ttacttgttc tgattctgtt tctggttctg
8401 gttctgttgg ctcatttggg attaaatcca ctactagcgt tgagttagtt aactttgcaa
8461 tttgttctag tgtttttatg gttggatctg attttcctga ttctattcgt gaataatttg
8521 atctactcat ttctaattct tggggtaccg ccagcatttc ggaaaaaaac cacgctaagg
8581 attttttcta taaaaagagc cgttatatta agaataaaac ggctctttta tacgtaaagg
8641 acgtaaattc atttgcccag tgtcatgtaa tccttcaaat ttgtattctc caagaaaatt
8701 gatatgttcc catcctaacg gccacgcata tggcattaaa tcttctctaa attctcctct
8761 tgcttttaat tcttctacgg ctttttccat atatacagtg ttccacacac ttatagcgtt
8821 aataattatg tttagtgcac tagctctttg taactggtct tggagagcac gttctctaaa
8881 ttctccacgt tgtccaaaaa atataattct agctaatgca ttgattgctt ctcctttatt
8941 taaacctttt tgaacccgtc tccttacggc tttattagat atgtaatcca gcgtaaagag
9001 ggttttctcg attcgtccca tttctccaag tgctgttgcg agtttatttt gtcttgcata
9061 tgatccgagc ttccccatga taagagcgct agggacctct ttagctcctt ggaagctgtc
9121 agtagtatac ctaataattt atctacattc cctttagtaa cgtgtaactt tccaaattta
9181 caaaagcgac tcatagaatt atttcctccc gttaaataat agataactat taaaaataga
9241 caatacttgc tcataagtaa cggtacttaa attgtttact ttggcgtgtt tcattgcttg
9301 atgaaactga tttttagtaa acagttgacg atattctcga ttgacccatt ttgaaacaaa
9361 gtacgtatat agcttccaat atttatctgg aacatctgtg gtatggcggg taagttttat
9421 taagacactg tttacttttg gtttaggatg aaagcattcc gctggcagct taagcaattg
9481 ctgaatcgag acttgagtgt gcaagagcaa ccctagtgtt cggtgaatat ccaaggtacg
9541 cttgtagaat ccttcttcaa caatcagata gatgtcagac gcatggcttt caaaaaccac
9601 ttttttaata atttgtgtgc ttaaatggta aggaatactc ccaacaattt tatacctctg
9661 tttgttaggg aattgaaact gtagaatatc ttggtgaatt aaagtgacac gagtattcag
9721 ttttaatttt tctgacgata agttgaatag atgactgtct aattcaatag acgttacctg
9781 tttacttatt ttagccagtt tcgtcgttaa atgcccttta cctgttccaa tttcgtaaac
9841 ggtatcggtt tcttttaaat tcaattgttt tattatttgg ttgagtactt tttcactcgt
9901 taaaaagttt tgagaatatt ttatattttt gttcatgtaa tcactccttc ttaattacaa
9961 atttttagca tctaatttaa cttcaattcc tattatacaa aattttaaga tactgcacta
10021 tcaacacact cttaagtttg cttctaagtc ttatttccat aacttctttt acgtttccgc
10081 cattctttgc tgtttcgatt tttatgatat ggtgcaagtc agcacgaaca cgaaccgtct
10141 tatctcccat tatatctttt tttgcactga ttggtgtatc atttcgtttt tctttttgtg
10201 cgcttcttga taaaagggat agtaattcat tcctggttgc aaattttgaa aaccgctacg
10261 gatcacatct ttttctaaac tagggcccat tagttgccga ctacgcgggc tgatacctgc
10321 taaaattgtt tgaaagtcgg gcgtttgctc aacaatcgtt tcaatatcca ttctgtcgtt
10381 tcctcccggg ctagttatat tgattcataa ctgccgtaaa atcatcattt gctaaccaag
10441 cttcgaccgc cgctagcgcc gtaatcttag catcgtcaaa cttcgccact tcggctggcg
10501 taaatttccc caacacgtaa tcaacaacac tcatcttctg aggatggtca ataccaactt
10561 taacgcgctt gaagtgctga tcaccaacat gactaataat gctcttgatg ccattatgac
10621 cacccgccga tcccttttgt ttgaggcgta cttttcctag tggtaaatct aaatcatcat
10681 gtataacgag caaatcagca ggatcaacat tgtaatagct catcaacggt cccaccgctc
10741 gtccagaatc attcatataa gtggcgggtt tcaccagtaa aatcttctca ccgtcttgaa
10801 aagccgttgc cacctgtgct tcaaatttac tcgtttgaaa cgtaacgttt agtttgctag
10861 ctaattcgtc cacaatcata aaaccaacat tatgacgtgt ctgggcatat tgccgcccaa
10921 tgttgcctaa cccaacgatc attttcataa ttttatcttc tcctattact ttgcatacca
10981 aaacaggccg aaccggtaat cgacccgatt cggcaactct gagtaacgat accacctaag
11041 tcgtattggc accactactc acaccgtgac cgacgcgccc gccagtcaag tgttcaaaag
11101 ttagcgttta ttaagtgcga taagtatacc acaaagggct tattgacgcc cgccaaaggg
11161 ttttgcggac attgttaata attgtattaa aagcatgctc aatctaacac ttattttgca
11221 caaacatggt atactttaac cgtaaaaact aaattttcac tacgagagga tgacttattt
11281 tgtcaagcaa taaataatca tttcatacga ttaaatgtat gatgaacgct cgtctatagc
11341 agacgggcgt ttttttgttt gcttgaggta ccttagcgat tcattaaagc gcaacacgca
11401 ctaaaggcta tttttaaaac tttcttatca cgattaccgg ccttgaagtt tgcactcatc
11461 tcacttctgt tataaggtga gaatattacg aatatatgga ggaccaactt aattatgaaa
11521 cataaacgtg gacttatgat tgggggcagt atcctcggtc tcctcgtcat cggatatgtt
11581 gccacgagct taatctggca acatcaacaa acgttcctca gtaatacgtc ggtcgctggt
11641 gttgacgtca gtggtaaaac cgccgctcaa gcagcaccga aagttaatcg cgctttagaa
11701 catcgcactt atcgaattat tgaaaacggc caaacaaagt actcatttac cagcaaatca
11761 gccggtatta caatcaatac taaaaaagat ttaaccaaat tagtgaccaa acaaaactac
11821 tggcgttggc cccttgcact attgcaatcg gctcaggctg atgatagctc agcggttgga
11881 caactaacca tcaatcacac tgatttgaaa accctactac aaaaaatcac cacgactgcc
11941 aatcaaacca aacgaacaaa aactgaaaac gccaagttag tttatcaaca taaccaagtt
12001 gagattaaga aagaagttca aggaaccgaa ttagatcaag ctaaacttaa gacggtcgtc
12061 accaaagcaa ttagtaatgg tgacagtcat atcaccctta aagacgccta cgtcactccg
12121 accgttacca gcacgagtgc tagtttaaag acggccaaag caaaatcagc gaaatacgct
12181 accgaaaagg ccacctataa tatcaacggt cataaattca cgattcctca cgacctaatt
12241 ctcagttgga ttaaagtcga taagcaaggc aaagtaagtt tagaccaact caaaaaaagc
12301 accgactcgg tgccactttt tcaagttgat aacggactag ccttatttta acttgctatt
12361 tctagctcta aaactagcga catcgcgaac tggatctaga atatatcata gtatgtccat
12421 tctgtcaaca actatagcgc tgtcgacgta tggacagttt tccctttgat atgtaacggt
12481 gaacagttgt tctacttttg tttgttagtc ttgatgcttc actgatagat acaagagcca
12541 taagaacctc agatccttcc gtatttagcc agtatgttct ctagtgtggt tcgttgtttt
12601 tgcgtgagcc atgagaacga accattgaga tcatacttac tttgcatgtc actcaaaaat
12661 tttgcctcaa aactggtgag ctgaattttt gcagttaaag catcgtgtag tgtttttctt
12721 agtccgttat gtaggtagga atctgatgta atggttgttg gtattttgtc accattcatt
12781 tttatctggt tgttctcaag ttcggttacg agatccattt gtctatctag ttcaacttgg
12841 aaaatcaacg tatcagtcgg gcggcctcgc ttatcaacca ccaatttcat attgctgtaa
12901 gtgtttaaat ctttacttat tggtttcaaa acccattggt taagcctttt aaactcatgg
12961 tagttatttt caagcattaa catgaactta aattcatcaa ggctaatctc tatatttgcc
13021 ttgtgagttt tcttttgtgt tagttctttt aataaccact cataaatcct catagagtat
13081 ttgttttcaa aagacttaac atgttccaga ttatatttta tgaatttttt taactggaaa
13141 agataaggca atatctcttc actaaaaact aattctaatt tttcgcttga gaacttggca
13201 tagtttgtcc actggaaaat ctcaaagcct ttaaccaaag gattcctgat ttccacagtt
13261 ctcgtcatca gctctctggt tgctttagct aatacaccat aagcattttc cctactgatg
13321 ttcatcatct gaacgtattg gttataagtg aacgataccg tccgttcttt ccttgtaggg
13381 ttttcaatcg tggggttgag tagtgccaca cagcataaaa ttagcttggt ttcatgctcc
13441 gttaagtcat agcgactaat cgctagttca tttgctttga aaacaactaa ttcagacata
13501 catctcaatt ggtctaggtg attttaatca ctataccaat tgagatgggc tagtcaatga
13561 taattactag tccttttcct ttgagttgtg ggtatctgta aattctgcta gacctttgct
13621 ggaaaacttg taaattctgc tagaccctct gtaaattccg ctagaccttt gtgtgttttt
13681 tttgtttata ttcaagtggt tataatttat agaataaaga aagaataaaa aaagataaaa
13741 agaatagatc ccagccctgt gtataactca ctactttagt cagttccgca gtattacaaa
13801 aggatgtcgc aaacgctgtt tgctcctcta caaaacagac cttaaaaccc taaaggctta
13861 agtagcaccc tcgcaagctc ggttgcggcc gcaatcgggc aaatcgctga atattccttt
13921 tgtctccgac catcaggcac ctgagtcgct gtctttttcg tgacattcag ttcgctgcgc
13981 tcacggctct ggcagtgaat gggggtaaat ggcactacag gcgcctttta tggattcatg
14041 caaggaaact acccataata caagaaaagc ccgtcacggg cttctcaggg cgttttatgg
14101 cg
//
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pHSP02质粒载体应用举例
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