| 质粒名称: | pRGE32 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 63159 |
| 存储实验室: | Yinong Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pRGE31 |
| 载体出品公司: | Kabin Xie |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | 用水稻UBI启动子+5'-UTR替换35S启动子。 |
| 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: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | Rice snoRNA U3 promoter for gRNA/PTG expression and UBI promoter for Cas9 expression |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | M13R(-48) |
| 3' 测序引物 | M13F(-20) |
|
文献引用方法 |
|
| 材料和方法部分: | pRGE32 was a gift from Yinong Yang (Addgene plasmid # 63159 ; http://n2t.net/addgene:63159 ; RRID:Addgene_63159) |
| 参考文献部分: | Boosting CRISPR/Cas9 multiplex editing capability with the endogenous tRNA-processing system. Xie K, Minkenberg B, Yang Y. Proc Natl Acad Sci U S A. 2015 Mar 17;112(11):3570-5. doi: 10.1073/pnas.1420294112. Epub 2015 Mar 2. 10.1073/pnas.1420294112 PubMed 25733849 |
买家导航
pRGE32载体质粒基本信息
pRGE32质粒图谱载体图谱和pRGE32载体序列质粒序列多克隆位点信息
pRGE32质粒载体简介
pRGE32质粒序列载体序列
LOCUS pRGE32 10092 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Express sgRNA/PTG with rice snoRNA U3 promoter and Cas9 with rice
ubiquitin promoter for transient expression in protoplast.
ACCESSION .
VERSION .
KEYWORDS pRGE32
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10092)
AUTHORS Xie K, Minkenberg B, Yang Y
TITLE Boosting CRISPR/Cas9 multiplex editing capability with the
endogenous tRNA-processing system.
JOURNAL Proc Natl Acad Sci U S A. 2015 Mar 17;112(11):3570-5. doi:
10.1073/pnas.1420294112. Epub 2015 Mar 2.
PUBMED 25733849
REFERENCE 2 (bases 1 to 10092)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10092
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 26..49
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
terminator 71..323
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(332..349)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(332..348)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(341..363)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 561..1016
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(648..667)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 858..879
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(1033..1052)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 1152..1174
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(1212..1230)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1298..1402
/gene="bla"
/label=AmpR promoter
CDS 1403..2263
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGTRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(1621..1640)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2434..3022
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2923..2942
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3176..3193
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 3310..3331
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3346..3376
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3384..3400
/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 3389..3411
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 3408..3424
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3408..3424
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 3443..3823
/label=OsU3 promoter
/note="Oryza sativa (rice) snRNA U3 promoter"
misc_RNA 3840..3915
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 3996..5705
/gene="RUBQ2"
/label=Ubi promoter
/note="promoter for the rice (Oryza sativa) polyubiquitin
gene RUBQ2"
intron 4744..5705
/note="intron upstream of the coding sequence for the rice
polyubiquitin gene RUBQ2"
protein_bind 5716..5740
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 5785..5850
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 5857..5877
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5902..10002
/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"
CDS 10003..10050
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
protein_bind complement(10083..15)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
ORIGIN
1 cttgtacaaa gtggttgata acagcgacta caaggatgac gatgacaagg cttagagctc
61 gaatttcccc gatcgttcaa acatttggca ataaagtttc ttaagattga atcctgttgc
121 cggtcttgcg atgattatca tataatttct gttgaattac gttaagcatg taataattaa
181 catgtaatgc atgacgttat ttatgagatg ggtttttatg attagagtcc cgcaattata
241 catttaatac gcgatagaaa acaaaatata gcgcgcaaac taggataaat tatcgcgcgc
301 ggtgtcatct atgttactag atcgggaatt cactggccgt cgttttacaa cgtcgtgact
361 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
421 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
481 gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
541 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
601 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
661 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
721 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
781 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
841 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
901 gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
961 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
1021 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
1081 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
1141 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
1201 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
1261 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
1321 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
1381 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
1441 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
1501 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
1561 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
1621 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
1681 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
1741 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
1801 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
1861 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
1921 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
1981 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
2041 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
2101 tggagccggt gagcgtggca ctcgcggtat cattgcagca ctggggccag atggtaagcc
2161 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
2221 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
2281 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
2341 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
2401 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
2461 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
2521 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
2581 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
2641 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
2701 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
2761 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
2821 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
2881 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
2941 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
3001 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
3061 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
3121 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
3181 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
3241 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
3301 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
3361 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
3421 tgaccatgat tacgccaagc ttaaggaatc tttaaacata cgaacagatc acttaaagtt
3481 cttctgaagc aacttaaagt tatcaggcat gcatggatct tggaggaatc agatgtgcag
3541 tcagggacca tagcacaaga caggcgtctt ctactggtgc taccagcaaa tgctggaagc
3601 cgggaacact gggtacgttg gaaaccacgt gatgtgaaga agtaagataa actgtaggag
3661 aaaagcattt cgtagtgggc catgaagcct ttcaggacat gtattgcagt atgggccggc
3721 ccattacgca attggacgac aacaaagact agtattagta ccacctcggc tatccacata
3781 gatcaaagct gatttaaaag agttgtgcag atgatccgtg gcaggagacc gaggtctcgg
3841 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
3901 gcaccgagtc ggtgcttttt tgttttagag ctagaaatag caagttaaaa taaggctagt
3961 ccgtttttag cgcgtgcatg cctgcaggtc cacaaattcg ggtcaaggcg gaagccagcg
4021 cgccacccca cgtcagcaaa tacggaggcg cggggttgac ggcgtcaccc ggtcctaacg
4081 gcgaccaaca aaccagccag aagaaattac agtaaaaaaa aagtaaattg cactttgatc
4141 caccttttat tacctaagtc tcaatttgga tcacccttaa acctatcttt tcaatttggg
4201 ccgggttgtg gtttggacta ccatgaacaa cttttcgtca tgtctaactt ccctttcagc
4261 aaacatatga accatatata gaggagatcg gccgtatact agagctgatg tgtttaaggt
4321 cgttgattgc acgagaaaaa aaaatccaaa tcgcaacaat agcaaattta tctggttcaa
4381 agtgaaaaga tatgtttaaa ggtagtccaa agtaaaactt atagataata aaatgtggtc
4441 caaagcgtaa ttcactcaaa aaaaatcaac gagacgtgta ccaaacggag acaaacggca
4501 tcttctcgaa atttcccaac cgctcgctcg cccgcctcgt cttcccggaa accgcggtgg
4561 tttcagcgtg gcggattctc caagcagacg gagacgtcac ggcacgggac tcctcccacc
4621 acccaaccgc cataaatacc agccccctca tctcctctcc tcgcatcagc tccacccccg
4681 aaaaatttct ccccaatctc gcgaggctct cgtcgtcgaa tcgaatcctc tcgcgtcctc
4741 aaggtacgct gcttctcctc tcctcgcttc gtttcgattc gatttcggac gggtgaggtt
4801 gttttgttgc tagatccgat tggtggttag ggttgtcgat gtgattatcg tgagatgttt
4861 aggggttgta gatctgatgg ttgtgatttg ggcacggttg gttcgatagg tggaatcgtg
4921 gttaggtttt gggattggat gttggttctg atgattgggg ggaattttta cggttagatg
4981 aattgttgga tgattcgatt ggggaaatcg gtgtagatct gttggggaat tgtggaacta
5041 gtcatgcctg agtgattggt gcgatttgta gcgtgttcca tcttgtaggc cttgttgcga
5101 gcatgttcag atctactgtt ccgctcttga ttgagttatt ggtgccatgg gttggtgcaa
5161 acacaggctt taatatgtta tatctgtttt gtgtttgatg tagatctgta gggtagttct
5221 tcttagacat ggttcaatta tgtagcttgt gcgtttcgat ttgatttcat atgttcacag
5281 attagataat gatgaactct tttaattaat tgtcaatggt aaataggaag tcttgtcgct
5341 atatctgtca taatgatctc atgttactat ctgccagtaa tttatgctaa gaactatatt
5401 agaatatcat gttacaatct gtagtaatat catgttacaa tctgtagttc atctatataa
5461 tctattgtgg taatttcttt ttactatctg tgtgaagatt attgccacta gttcattcta
5521 cttatttctg aagttcagga tacgtgtgct gttactacct atctgaatac atgtgtgatg
5581 tgcctgttac tatctttttg aatacatgta tgttctgttg gaatatgttt gctgtttgat
5641 ccgttgttgt gtccttaatc ttgtgctagt tcttacccta tctgtttggt gattatttct
5701 tgcagatagt tatcaacaag tttgtacaaa aaagcaggct tcgaaggaga tagaaccaat
5761 tctctaagga aatacttaac catggactat aaggaccacg acggagacta caaggatcat
5821 gatattgatt acaaagacga tgacgataag atggccccaa agaagaagcg gaaggtcggt
5881 atccacggag tcccagcagc cgacaagaag tacagcatcg gcctggacat cggcaccaac
5941 tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg
6001 ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac
6061 agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga
6121 cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac
6181 gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag
6241 cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc
6301 atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc
6361 tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg
6421 aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag
6481 ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc
6541 agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag
6601 aatggcctgt tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc
6661 aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac
6721 ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag
6781 aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag
6841 gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg
6901 ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag
6961 agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag
7021 ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac
7081 agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc
7141 cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag
7201 gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct
7261 ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc
7321 ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg
7381 atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg
7441 tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg
7501 agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag
7561 accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc
7621 ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac
7681 cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac
7741 attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa
7801 cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg
7861 agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag
7921 tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg
7981 cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc
8041 ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag
8101 aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac
8161 aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag
8221 aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag
8281 atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac
8341 tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc
8401 gactacgatg tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac
8461 aaggtgctga ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag
8521 gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag
8581 agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc
8641 ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc
8701 ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa
8761 gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa
8821 gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga
8881 accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag
8941 gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc
9001 aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac
9061 ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg
9121 gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc
9181 gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg
9241 aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc
9301 gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc
9361 aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc
9421 gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg
9481 atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg
9541 gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac
9601 ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag
9661 aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc
9721 gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac
9781 aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc
9841 ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag
9901 aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc
9961 ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acaaaaggcc ggcggccacg
10021 aaaaaggccg gccaggcaaa aaagaaaaag taagaattcg cggccgcact cgagatatct
10081 agacccagct tt
//
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pRGE32质粒载体应用举例
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