| 质粒名称: | pTSin EGFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 127695 |
| 存储实验室: | Justin English, Bryan Roth |
|
载体骨架基本信息 |
|
| 载体名称: | pSin |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | Synthetic Biology |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | EGFP |
| 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: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pTSin EGFP was a gift from Justin English & Bryan Roth (Addgene plasmid # 127695 ; http://n2t.net/addgene:127695 ; RRID:Addgene_127695) |
| 参考文献部分: | VEGAS as a Platform for Facile Directed Evolution in Mammalian Cells. English JG, Olsen RHJ, Lansu K, Patel M, White K, Cockrell AS, Singh D, Strachan RT, Wacker D, Roth BL. Cell. 2019 Jul 2. pii: S0092-8674(19)30622-1. doi: 10.1016/j.cell.2019.05.051. 10.1016/j.cell.2019.05.051 PubMed 31280962 |
买家导航
pTSinEGFP载体基本信息
pTSinEGFP载体图谱质粒图谱和多克隆位点信息
pTSinEGFP载体简介
pTSinEGFP载体序列
LOCUS pTSin EGFP 10667 bp ds-DNA circular SYN 03-JUL-2019
DEFINITION Plasmid contains the entire replication competent Sindbis genome
with the structural genome components replaced by EGFP in an MCS
locus..
ACCESSION .
VERSION .
KEYWORDS pTSin EGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10667)
TITLE Roth Lab Sindbis Plasmids
REFERENCE 2 (bases 1 to 10667)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jul 3, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10667
/organism="synthetic DNA construct"
/mol_type="other DNA"
regulatory 7682..7691
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 7688..8407
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(7733..7754)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(7994..8013)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 8341..8362
/label=EGFP-C
/note="EGFP, forward primer"
promoter 8838..8942
/gene="bla"
/label=AmpR promoter
CDS 8943..9803
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(9161..9180)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 9974..10562
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 10463..10482
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 10650..1
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 10650..10667
/label=SP6
/note="SP6 promoter, forward primer"
ORIGIN
1 attggcggcg tagtacacac tattgaatca aacagccgac caattgcact accatcacaa
61 tggagaagcc agtagttaac gtagacgtag acccgcagag tccgtttgtc gtgcaactgc
121 aaaagagctt cccgcaattt gaggtagtag cacagcaggt cactccaaat gaccatgcta
181 atgccagagc attttcgcat ctggccagta aactaatcga gctggaggtt cctaccacag
241 cgacgatttt ggacataggc agcgcaccgg ctcgtagaat gttttccgag caccagtacc
301 attgcgtttg ccccatgcgt agtccagaag acccggaccg catgatgaaa tatgccagca
361 aactggcgga aaaagcatgc aagattacga ataagaactt gcatgagaag atcaaggacc
421 tccggaccgt acttgataca ccggatgctg aaacgccatc actctgcttc cacaacgatg
481 ttacctgcaa cacgcgtgcc gagtactccg tcatgcagga cgtgtacatc aacgcacccg
541 gaactattta ccatcaggct atgaaaggcg tgcggaccct gtactggatt ggcttcgata
601 ccacccagtt catgttctcg gctatggcag gttcgtaccc tgcgtacaac accaactggg
661 ccgacgaaaa agtcctcgaa gcgcgtaaca tcggactctg cagcacaaag ctgagtgaag
721 gcaggacagg aaagttgtcg ataatgagga agaaggagtt gaagcccggg tcacgggttt
781 atttctccgt tggatcgaca ctttacccag aacacagagc cagcttgcag agctggcatc
841 ttccatcggt gttccacctg aaaggaaagc agtcgtacac ttgccgctgt gatacagtgg
901 tgagctgcga aggctacgta gtgaagaaaa tcaccatcag tcccgggatc acgggagaaa
961 ccgtgggata cgcggttaca aacaatagcg agggcttctt gctatgcaaa gttaccgata
1021 cagtaaaagg agaacgggta tcgttccccg tgtgcacgta tatcccggcc accatatgcg
1081 atcagatgac cggcataatg gccacggata tctcacctga cgatgcacaa aaacttctgg
1141 ttgggctcaa ccagcgaatc gtcattaacg gtaagactaa caggaacacc aataccatgc
1201 aaaattacct tctgccaatc attgcacaag ggttcagcaa atgggccaag gagcgcaaag
1261 aagaccttga caatgaaaaa atgctgggta ccagagagcg caagcttaca tatggctgct
1321 tgtgggcgtt tcgcactaag aaagtgcact cgttctatcg cccacctgga acgcagacca
1381 tcgtaaaagt cccagcctct tttagcgctt tccccatgtc atccgtatgg actacctctt
1441 tgcccatgtc gctgaggcag aagataaaat tggcattaca accaaagaag gaggaaaaac
1501 tgctgcaagt cccggaggaa ttagtcatgg aggccaaggc tgctttcgag gatgctcagg
1561 aggaatccag agcggagaag ctccgagaag cactcccacc attagtggca gacaaaggta
1621 tcgaggcagc cgcggaagtt gtctgcgaag tggaggggct ccaggcggac atcggagcag
1681 cactcgtcga aaccccgcgc ggtcatgtaa ggataatacc tcaagcaaat gaccgtatga
1741 tcggacagta catcgttgtc tcgccaacct ctgtgctgaa gaacgctaaa ctcgcaccag
1801 cacacccgct agcagaccag gttaagatca taacgcactc cggaagatca ggaaggtatg
1861 cagtcgaacc atacgacgct aaagtactga tgccagcagg aagtgccgta ccatggccag
1921 aattcttagc actgagtgag agcgccacgc tagtgtacaa cgaaagagag tttgtgaacc
1981 gcaagctgta ccatattgcc atgcacggtc ccgctaagaa tacagaagag gagcagtaca
2041 aggttacaaa ggcagagctc gcagaaacag agtacgtgtt tgacgtggac aagaagcgat
2101 gcgtcaagaa ggaagaagcc tcaggacttg tcctctcggg agaactgacc aacccgccct
2161 atcacgaact agctcttgag ggactgaaga ctcgacccgc ggtcccgtac aaggttgaaa
2221 caataggagt gataggcaca ccaggatcgg gcaagtcggc tatcatcaag tcaactgtca
2281 cggcacgtga tcttgttacc agcggaaaga aagaaaactg ccgcgaaatt gaggccgatg
2341 tgctacggct gaggggcatg cagatcacgt cgaagacagt ggattcggtt atgctcaacg
2401 gatgccacaa agccgtagaa gtgctgtatg ttgacgaagc gttcgcgtgc cacgcaggag
2461 cactacttgc cttgattgca atcgtcagac cccgtaagaa ggtagtgcta tgcggagacc
2521 ctaagcaatg cggattcttc aacatgatgc aactaaaggt atatttcaac cacccggaaa
2581 aagacatatg taccaagaca ttctacaagt ttatctcccg acgttgcaca cagccagtca
2641 cggctattgt atcgacactg cattacgatg gaaaaatgaa aaccacaaac ccgtgcaaga
2701 agaacatcga aatcgacatt acaggggcca cgaagccgaa gccaggggac atcatcctga
2761 catgcttccg cgggtgggtt aagcaactgc aaatcgacta tcccggacat gaggtaatga
2821 cagccgcggc ctcacaaggg ctaaccagaa aaggagtata tgccgtccgg caaaaagtca
2881 atgaaaaccc gctgtacgcg atcacatcag agcatgtgaa cgtgctgctc acccgcactg
2941 aggacaggct agtatggaaa actttacagg gcgacccatg gattaagcag ctcactaacg
3001 taccaaaagg aaattttcaa gccaccatcg aggactggga agctgaacac aagggaataa
3061 ttgctgcgat aaacagtccc gctccccgta ccaatccgtt cagctgcaag actaacgttt
3121 gctgggcgaa agcactggaa ccgatactgg ccacggccgg tatcgtactt accggttgcc
3181 agtggagcga gctgttccca cagtttgcag atgacaaacc acactcggcc atctacgccc
3241 tggacgtaat ctgcattaag tttttcggca tggacttgac aagcggactg ttttccaaac
3301 agagcatccc gttaacgtac catcctgccg attcagcgag gccagtagct cattgggaca
3361 acagcccagg aacccgcaag tatgggtacg atcacgccgt tgccgccgaa ctctcccgta
3421 gatttccggt gttccagcta gctgggaaag gcacacagct tgatttgcag acgggcagaa
3481 ctagagttat ctccgcacag cataacttgg tcccagtgaa ccgcaatctc ccgcacgcct
3541 tagtccccga gcacaaggag aaacaacccg gcccggtcaa aaaattcttg agccagttca
3601 aacaccactc cgtacttgtg gtctcagagg aaaaaattga agctccccac aagagaatcg
3661 aatggatcgc cccgattggc atagccggcg ctgataagaa ctacaacctg gctttcgggt
3721 ttccgccgca ggcacggtac gacctggtgt ttatcaatat tggaactaaa tacagaaacc
3781 atcactttca gcagtgcgaa gaccatgcgg cgaccttgaa aaccctctcg cgttcggccc
3841 tgaactgcct taaccccgga ggcaccctcg tggtgaagtc ctacggttac gccgaccgca
3901 atagtgagga cgtagtcacc gctcttgcca gaaaatttgt cagagtgtct gcagcgaggc
3961 cagagtgcgt ctcaagcaat acagaaatgt acctgatctt ccgacaacta gacaacagcc
4021 gcacacgaca attcaccccg catcatctga attgtgtgat ttcgtccgtg tacgagggta
4081 caagagacgg agttggagcc gcaccgtcat accgcactaa aagggagaac attgctgatt
4141 gtcaagagga agcagttgtc aatgcagcca atccgctggg cagaccaggc gaaggagtct
4201 gccgtgccat ctataaacgt tggccgaaca gtttcaccga ttcagccaca gagaccggca
4261 ccgcaaaact gactgtgtgc caaggaaaga aagtgatcca cgcggttggc cctgatttcc
4321 ggaaacaccc agaggcagaa gccctgaaat tgctgcaaaa cgcctaccat gcagtggcag
4381 acttagtaaa tgaacataat atcaagtctg tcgccatccc actgctatct acaggcattt
4441 acgcagccgg aaaagaccgc cttgaagtat cacttaactg cttgacaacc gcgctagata
4501 gaactgatgc ggacgtaacc atctactgcc tggataagaa gtggaaggaa agaatcgacg
4561 cggtgctcca acttaaggag tctgtaacag agctgaagga tgaggatatg gagatcgacg
4621 acgagttagt atggatccat ccggacagtt gcctgaaggg aagaaaggga ttcagtacta
4681 caaaaggaaa gttgtattcg tactttgaag gcaccaaatt ccatcaagca gcaaaagata
4741 tggcggagat aaaggtcctg ttcccaaatg accaggaaag caacgagcaa ctgtgtgcct
4801 acatattggg ggagaccatg gaagcaatcc gcgaaaaatg cccggtcgac cacaacccgt
4861 cgtctagccc gccaaaaacg ctgccgtgcc tctgcatgta tgccatgacg ccagaaaggg
4921 tccacagact cagaagcaac aacgtcaaag aagttacagt atgctcctcc accccccttc
4981 caaagtacaa aatcaagaac gttcagaagg ttcagtgcac aaaagtagtc ctgtttaacc
5041 cgcatacccc tgcattcgtt cccgcccgta agtacataga agcgccagaa cagcctgcag
5101 ctccgcctgc acaggccgag gaggcccccg aagttgcagc aacaccaaca ccacctgcag
5161 ctgataacac ctcgcttgat gtcacggaca tctcactgga catggaagac agtagcgaag
5221 gctcactctt ttcgagcttt agcggatcgg acaactctat tactagtatg gacagttggt
5281 cgtcaggacc tagttcacta gagatagtag accgaaggca ggtggtggtg gctgacgtcc
5341 atgccgtcca agagcctgcc cctgttccac cgccaaggct aaagaagatg gcccgcctgg
5401 cagcggcaag aatgcaggaa gagccaactc caccggcaag caccagctct gcggacgagt
5461 cccttcacct ttcttttggt ggggtatcca tgtccttcgg atcccttttc gacggagaga
5521 tggcccgctt ggcagcggca caacccccgg caagtacatg ccctacggat gtgcctatgt
5581 ctttcggatc gttttccgac ggagagattg aggagctgag ccgcagagta accgagtctg
5641 agcccgtcct gtttgggtca tttgaaccgg gcgaagtgaa ctcaattata tcgtcccgat
5701 cagccgtatc ttttccacca cgcaagcaga gacgtagacg caggagcagg aggaccgaat
5761 actgactaac cggggtaggt gggtacatat tttcgacgga cacaggccct gggcacttgc
5821 aaaagaagtc cgttctgcag aaccagctta cagaaccgac cttggagcgc aatgttctgg
5881 aaagaatcta cgccccggtg ctcgacacgt cgaaagagga acagctcaaa ctcaggtacc
5941 agatgatgcc caccgaagcc aacaaaagca ggtaccagtc cagaaaagta gaaaatcaga
6001 aagccataac cactgagcga ctgctttcag ggctacgact gtataactct gccacagatc
6061 agccagaatg ctataagatc acctacccga aaccatcgta ttccagcagt gtaccggcga
6121 actactctga cccaaagttt gctgtagctg tttgcaacaa ctatctgcat gagaattacc
6181 cgacggtagc atcttatcag atcaccgacg agtacgatgc ttacttggat atggtagacg
6241 ggacagtcgc ttgcctagat actgcaactt tttgccccgc caagcttaga agttacccga
6301 aaagacacga gtatagagcc ccaaacatcc gcagtgcggt tccatcagcg atgcagaaca
6361 cgttgcaaaa cgtgctcatt gccgcgacta aaagaaactg caacgtcaca caaatgcgtg
6421 aattgccaac actggactca gcgacattca acgttgaatg ctttcgaaaa tatgcatgta
6481 atgacgagta ttgggaggag tttgcccgaa agccaattag gatcactact gagttcgtta
6541 ccgcatacgt ggccagactg aaaggcccta aggccgccgc actgttcgca aagacgcata
6601 atttggtccc attgcaagaa gtgcctatgg ataggttcgt catggacatg aaaagagacg
6661 tgaaagttac acctggcacg aaacacacag aagaaagacc gaaagtacaa gtgatacaag
6721 ccgcagaacc cctggcgacc gcttacctgt gcgggatcca ccgggagtta gtgcgcaggc
6781 ttacagccgt cttgctaccc aacattcaca cgctttttga catgtcggcg gaggactttg
6841 atgcaatcat agcagaacac ttcaagcaag gtgacccggt actggagacg gatatcgcct
6901 cgttcgacaa aagccaagac gacgctatgg cgttaactgg cctgatgatc ttggaagacc
6961 tgggtgtgga ccaaccacta ctcgacttga tcgagtgcgc ctttggagaa atatcatcca
7021 cccatctgcc cacgggtacc cgtttcaaat tcggggcgat gatgaaatcc ggaatgttcc
7081 tcacgctctt tgtcaacaca gttctgaatg tcgttatcgc cagcagagta ttggaggagc
7141 ggcttaaaac gtccaaatgt gcagcattta tcggcgacga caacatcata cacggagtag
7201 tatctgacaa agaaatggct gagaggtgtg ccacctggct caacatggag gttaagatca
7261 ttgacgcagt catcggcgag agaccgcctt acttctgcgg tggattcatc ttgcaagatt
7321 cggttacctc cacagcgtgt cgcgtggcgg accccttgaa aaggctgttt aagttgggta
7381 aaccgctccc agccgacgat gagcaagacg aagacagaag acgcgctctg ctagatgaaa
7441 caaaggcgtg gtttagagta ggtataacag acaccttagc agtggccgtg gcaactcggt
7501 atgaggtaga caacatcaca cctgtcctgc tggcattgag aacttttgcc cagagcaaaa
7561 gagcatttca agccatcaga ggggaaataa agcatctcta cggtggtcct aaatagtcag
7621 catagtacat ttcatctgac taataccaca acaccaccag cggccgcatc gatcaccggt
7681 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
7741 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
7801 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
7861 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
7921 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
7981 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
8041 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
8101 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
8161 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
8221 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
8281 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
8341 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
8401 caagtaaagc ggcatcgata ctagtacgcc ccaatgaccc gaccagcaaa actcgatgta
8461 cttccgagga actgatgtgc ataatgcatc aggctggtat attagatccc cgcttaccgc
8521 gggcaatata gcaacaccaa aactcgacgt atttccgagg aggcgcagtg cataatgctg
8581 cgcagtgttg ccaaataatc actatattaa ccatttattc agcggacgcc aaaactcaat
8641 gtatttctga ggaagcatgg tgcataatgc catgcagcgt ctgcataact ttttattatt
8701 tcttttatta atcaacaaaa ttttgttttt aacatttcaa aaaaaaaaaa aaaaaaaggg
8761 gatctcgagc tctagagctc gaggggaatt aattcttgaa gacgaaaggg ccaggtggca
8821 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
8881 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
8941 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
9001 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
9061 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
9121 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
9181 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
9241 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
9301 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
9361 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
9421 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
9481 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
9541 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
9601 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
9661 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
9721 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
9781 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
9841 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
9901 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
9961 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
10021 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
10081 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
10141 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
10201 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
10261 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
10321 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
10381 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
10441 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
10501 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
10561 aaaacgccag caacgcgagc tcgtatggac atattgtcgt tagaacgcgg ctacaattaa
10621 tacataacct tatgtatcat acacatacga tttaggtgac actatag
//
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pTSinEGFP载体质粒应用举例
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