| 质粒名称: | pCS6-StaPLd-Casp9 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 111511 |
| 存储实验室: | Michael Lin |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV-SPORT6 |
| 载体出品公司: | Invitrogen |
| 空载体大小: | -- |
| 完整质粒大小: | 6801 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | StaPLd-Casp9 |
| 物种来源: | H. sapiens (human); Hepatitis C Virus (HCV) genotype 1a |
| 插入大小: | 2457 bp |
| 基因突变: | HCV NS3蛋白酶携带V36M、T54A和S122G突变。两个胱天蛋白酶-9拷贝的N末端都被截短了。 |
| GenBank编号 | NC_004102 |
| 启动子: | CMV |
| 标签/融合蛋白: | -- |
| 克隆方法: | Gibson Cloning |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | ACAGCTATGACCATTAGGCCT |
| 3’测序引物 | GTTGTAAAACGACGGCCAGT |
|
文献引用方法 |
|
| 材料和方法部分: | pCS6-StaPLd-Casp9 was a gift from Michael Lin (Addgene plasmid # 111511 ; http://n2t.net/addgene:111511 ; RRID:Addgene_111511) |
| 参考文献部分: | StaPLs: versatile genetically encoded modules for engineering drug-inducible proteins. Jacobs CL, Badiee RK, Lin MZ. Nat Methods. 2018 Jul;15(7):523-526. doi: 10.1038/s41592-018-0041-z. Epub 2018 Jul 2. 10.1038/s41592-018-0041-z PubMed 29967496 |
买家导航
pCS6-StaPLd-Casp9载体质粒基本信息
pCS6-StaPLd-Casp9质粒图谱载体图谱和pCS6-StaPLd-Casp9载体序列质粒序列多克隆位点信息
pCS6-StaPLd-Casp9质粒载体简介
pCS6-StaPLd-Casp9质粒序列载体序列
LOCUS pCS6-StaPLd-Casp9 6798 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses a tandem caspase-9 dimer, governed by a StaPL(AI) module,
in mammalian cells. [AI = asunaprevir inhibited]..
ACCESSION .
VERSION .
KEYWORDS pCS6-StaPLd-Casp9
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6798)
AUTHORS Jacobs CL, Badiee RK, Lin MZ
TITLE StaPLs: versatile genetically encoded modules for engineering
drug-inducible proteins.
JOURNAL Nat Methods. 2018 Jul;15(7):523-526. doi: 10.1038/s41592-018-0041-z.
Epub 2018 Jul 2.
PUBMED 29967496
REFERENCE 2 (bases 1 to 6798)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6798
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 157..1017
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(375..394)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
protein_bind complement(1119..1152)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
rep_origin 1230..1818
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 1719..1738
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1972..1989
/label=L4440
/note="L4440 vector, forward primer"
enhancer 2277..2580
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2581..2784
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 2735..2755
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 2781..2805
/label=LNCX
/note="Human CMV promoter, forward primer"
primer_bind 2910..2926
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2910..2926
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 2937..2955
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 2937..2954
/label=SP6
/note="SP6 promoter, forward primer"
protein_bind 2956..2980
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
regulatory 2999..3008
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS complement(3326..3337)
/codon_start=1
/product="WELQut protease recognition and cleavage site"
/label=WELQut site
/translation="WELQ"
CDS 3872..3898
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS complement(4943..4954)
/codon_start=1
/product="WELQut protease recognition and cleavage site"
/label=WELQut site
/translation="WELQ"
protein_bind complement(5478..5502)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
primer_bind complement(5503..5522)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(5504..5522)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5533..5550)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(5533..5549)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5542..5564)
/label=M13/pUC Forward
/note="In lacZ gene"
polyA_signal 5954..6088
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5991..6010)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6045..6064
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 6214..6668
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6301..6320)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6510..6531
/label=F1ori-F
/note="F1 origin, forward primer"
ORIGIN
1 gctggaggga gaataaaggt ctaagatgtg cgatagaggg aagtcgcatt gaattatgtg
61 ctgtgtaggg atcgctggta tcaaatatgt gtgcccaccc ctggcatgag acaataaccc
121 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
181 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
241 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
301 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
361 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
421 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
481 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
541 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
601 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
661 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
721 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
781 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
841 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
901 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
961 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
1021 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
1081 aggatctagg tgaagatcct ttttgataat ctcatgccat aacttcgtat aatgtatgct
1141 atacgaagtt atggcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
1201 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
1261 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
1321 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
1381 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
1441 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
1501 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
1561 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
1621 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
1681 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
1741 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
1801 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
1861 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
1921 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
1981 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
2041 attcattaat gcagagcttg caattcgcgc tttttcaata ttattgaagc atttatcagg
2101 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
2161 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
2221 cattaaccta taaaaatagg cgtattacga ggccctttca cgcattagat gcatgtcgtt
2281 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
2341 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
2401 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
2461 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
2521 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
2581 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
2641 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
2701 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
2761 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
2821 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg gactctagcc
2881 taggccgcgg gacggataac aatttcacac aggaaacagc tatgaccatt aggcctattt
2941 aggtgacact atagaacaag tttgtacaaa aaagcaggct ggtaccggtc cggaattcgc
3001 caccatggat gtcggtgctc ttgagagttt gaggggaaat gcagatttgg cttacatcct
3061 gagcatggag ccctgtggcc actgcctcat tatcaacaat gtgaacttct gccgtgagtc
3121 cgggctccgc acccgcactg gctccaacat cgactgtgag aagttgcggc gtcgcttctc
3181 ctcgctgcat ttcatggtgg aggtgaaggg cgacctgact gccaagaaaa tggtgctggc
3241 tttgctggag ctggcgcggc aggaccacgg tgctctggac tgctgcgtgg tggtcattct
3301 ctctcacggc tgtcaggcca gccacctgca gttcccaggg gctgtctacg gcacagatgg
3361 atgccctgtg tcggtcgaga agattgtgaa catcttcaat gggaccagct gccccagcct
3421 gggagggaag cccaagctct ttttcatcca ggcctgtggt ggggagcaga aagaccatgg
3481 gtttgaggtg gcctccactt cccctgaaga cgagtcccct ggcagtaacc ccgagccaga
3541 tgccaccccg ttccaggaag gtttgaggac cttcgaccag ctggacgcca tatctagttt
3601 gcccacaccc agtgacatct ttgtgtccta ctctactttc ccaggttttg tttcctggag
3661 ggaccccaag agtggctcct ggtacgttga gaccctggac gacatctttg agcagtgggc
3721 tcactctgaa gacctgcagt ccctcctgct tagggtcgct aatgctgttt cggtgaaagg
3781 gatttataaa cagatgcctg gttgctttaa tttcctccgg aaaaaacttt tctttaaaac
3841 atcagggggc agtggcggga gcgggcgcgc ctatccctac gatgtgcccg attacgctgg
3901 tggatccggc gggtcagatg agatggaaga gtgctctcag cacggcggat ctggaggttc
3961 tacaggctgc gtggtcatag tgggcaggat cgtcttgtcc ggatccggca ctagtgcgcc
4021 catcacggcg tacgcccagc agacgagagg cctcctaggg tgtataatca ccagcctgac
4081 tggccgggac aaaaaccaag tggagggtga ggtccagatc atgtcaactg ctacccaaac
4141 cttcctggca acgtgcatca atggggtatg ctgggcagtc taccacgggg ccggaacgag
4201 gaccatcgca tcacccaagg gtcctgtcat ccagatgtat accaatgtgg accaagacct
4261 tgtgggctgg cccgctcctc aaggttcccg ctcattgaca ccctgtacct gcggctcctc
4321 ggacctttac ctggtcacga ggcacgccga tgtcattccc gtgcgccggc gaggtgatgg
4381 caggggtagc ctgctttcgc cccggcccat ttcctacttg aaaggctcct cggggggtcc
4441 gctgttgtgc cccgcgggac acgccgtggg cctattcagg gccgcggtgt gcacccgtgg
4501 agtggctaaa gcggtggact ttatccctgt ggagaaccta gagacaacca tgagatcccc
4561 ggtgttcacg gacaactcct ctccaccagc agtcaccctg acgcacgggg gttcaggcgg
4621 tagtgatgtc ggtgctcttg agagtttgag gggaaatgca gatttggctt acatcctgag
4681 catggagccc tgtggccact gcctcattat caacaatgtg aacttctgcc gtgagtccgg
4741 gctccgcacc cgcactggct ccaacatcga ctgtgagaag ttgcggcgtc gcttctcctc
4801 gctgcatttc atggtggagg tgaagggcga cctgactgcc aagaaaatgg tgctggcttt
4861 gctggagctg gcgcggcagg accacggtgc tctggactgc tgcgtggtgg tcattctctc
4921 tcacggctgt caggccagcc acctgcagtt cccaggggct gtctacggca cagatggatg
4981 ccctgtgtcg gtcgagaaga ttgtgaacat cttcaatggg accagctgcc ccagcctggg
5041 agggaagccc aagctctttt tcatccaggc ctgtggtggg gagcagaaag accatgggtt
5101 tgaggtggcc tccacttccc ctgaagacga gtcccctggc agtaaccccg agccagatgc
5161 caccccgttc caggaaggtt tgaggacctt cgaccagctg gacgccatat ctagtttgcc
5221 cacacccagt gacatctttg tgtcctactc tactttccca ggttttgttt cctggaggga
5281 ccccaagagt ggctcctggt acgttgagac cctggacgac atctttgagc agtgggctca
5341 ctctgaagac ctgcagtccc tcctgcttag ggtcgctaat gctgtttcgg tgaaagggat
5401 ttataaacag atgcctggtt gctttaattt cctccggaaa aaacttttct ttaaaacatc
5461 atagaagctt tacgcgtacc cagctttctt gtacaaagtg gtccctatag tgagtcgtat
5521 tataagctag gcactggccg tcgttttaca acgtcgtgac tgggaaaact gctagcttgg
5581 gatctttgtg aaggaacctt acttctgtgg tgtgacataa ttggacaaac tacctacaga
5641 gatttaaagc tctaaggtaa atataaaatt tttaagtgta taatgtgtta aactagctgc
5701 atatgcttgc tgcttgagag ttttgcttac tgagtatgat ttatgaaaat attatacaca
5761 ggagctagtg attctaattg tttgtgtatt ttagattcac agtcccaagg ctcatttcag
5821 gcccctcagt cctcacagtc tgttcatgat cataatcagc cataccacat ttgtagaggt
5881 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
5941 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
6001 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
6061 catcaatgta tcttatcatg tctggatcga tcctgcatta atgaatcggc caacgcgcgg
6121 ggagaggcgg tttgcgtatt ggctggcgta atagcgaaga ggcccgcacc gatcgccctt
6181 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
6241 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
6301 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
6361 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
6421 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
6481 ctttgacgtt ggagtccacg ttcttaatag tggactcttg ttccaaactg gaacaacact
6541 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
6601 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
6661 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
6721 taaatacatt caaatatgta tccgctcatg ccaggtcttg gactggtgag aacggcttgc
6781 tcggcagctt cgatgtgt
//
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