| 质粒名称: | pGL3-NFAT luciferase |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 17870 |
| 存储实验室: | Jerry Crabtree |
|
载体骨架基本信息 |
|
| 载体名称: | pGL3 |
| 载体出品公司: | Promega |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 4800 bp |
| 载体类型: | 哺乳细胞表达，荧光素酶 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
| 宿主菌: | DH5a |
|
插入基因信息 |
|
| 插入基因名称: | 3x NFAT binding sequence |
| 基因又名: | NF-AT site |
| Enterz Gene: | IL2 (a.k.a. IL-2, TCGF, lymphokine) |
| 启动子: | IL-2启动子 |
| 标签/融合蛋白: | C-luciferase |
| 克隆方法: | 多克隆位点 |
| 5’测序引物 | RVprimer3 (5'-CTAGCAAAATAGGCTGTCCC-3') |
| 3’测序引物 | LucNrev |
|
文献引用方法 |
|
| 材料和方法部分: | pGL3-NFAT luciferase was a gift from Jerry Crabtree (Addgene plasmid # 17870 ; http://n2t.net/addgene:17870 ; RRID:Addgene_17870) |
| 参考文献部分: |
Identification of calcineurin as a key signalling enzyme in T-lymphocyte activation. Clipstone NA, Crabtree GR. Nature. 1992 Jun 25. 357(6380):695-7. 10.1038/357695a0 PubMed 1377362 |
买家导航
pGL3-NFAT luciferase载体质粒基本信息
pGL3-NFAT luciferase质粒图谱载体图谱和pGL3-NFAT luciferase载体序列质粒序列多克隆位点信息
pGL3-NFAT luciferase质粒载体简介
pGL3-NFAT luciferase质粒序列载体序列
LOCUS pGL3-NFAT luciferase 5049 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION 3x NFAT binding sequence upstream of a luciferase reporter; can be
used to assay signal transduction through Ca2+, calcineurin and NFAT
in any tissue.
ACCESSION .
VERSION .
KEYWORDS pGL3-NFAT luciferase
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5049)
AUTHORS Clipstone NA, Crabtree GR
TITLE Identification of calcineurin as a key signalling enzyme in
T-lymphocyte activation.
JOURNAL Nature. 1992 Jun 25. 357(6380):695-7.
PUBMED 1377362
REFERENCE 2 (bases 1 to 5049)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5049
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 1105..1123
/label=LucNrev
/note="Firefly luciferase, reverse primer"
regulatory 1186..1195
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
promoter complement(1227..1340)
/label=minimal IL-2 promoter
/label=IL-2 promoter
/note="minimal promoter from the human interleukin-2 gene"
protein_bind 1348..1377
/label=NFAT binding site
/bound_moiety="NFAT transcription factor"
/note="NFAT binding site from the human interleukin 2
(IL-2) gene (Mattila et al., 1990)"
protein_bind 1378..1407
/label=NFAT binding site
/bound_moiety="NFAT transcription factor"
/note="NFAT binding site from the human interleukin 2
(IL-2) gene (Mattila et al., 1990)"
protein_bind 1408..1437
/label=NFAT binding site
/bound_moiety="NFAT transcription factor"
/note="NFAT binding site from the human interleukin 2
(IL-2) gene (Mattila et al., 1990)"
misc_feature 1515..1606
/label=pause site
/note="RNA polymerase II transcriptional pause signal from
the human alpha-2 globin gene"
primer_bind complement(1547..1566)
/label=RVprimer3
/note="pGL3 vector, forward primer"
polyA_signal 1620..1668
/note="synthetic polyadenylation signal"
rep_origin complement(1799..2254)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1936..1957)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2148..2167
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2281..2385
/gene="bla"
/label=AmpR promoter
CDS 2386..3246
/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(2604..2623)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3417..4005
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3906..3925
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4159..4176
/label=L4440
/note="L4440 vector, forward primer"
polyA_signal 4424..4545
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4461..4480)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4515..4534
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
CDS complement(4586..1189)
/codon_start=1
/gene="luc+"
/product="firefly luciferase"
/label=luciferase
/note="enhanced luc+ version of the luciferase gene"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
ORIGIN
1 tcataggacc tctcacacac agttcgcctc tttgattaac gcccagcgtt ttcccggtat
61 ccagatccac aaccttcgct tcaaaaaatg gaacaacttt accgaccgcg cccggtttat
121 catccccctc gggtgtaatc agaatagctg atgtagtctc agtgagccca tatccttgcc
181 tgatacctgg cagatggaac ctcttggcaa ccgcttcccc gacttcctta gagaggggag
241 cgccaccaga agcaatttcg tgtaaattag ataaatcgta tttgtcaatc agagtgcttt
301 tggcgaagaa ggagaatagg gttggcacca gcagcgcact ttgaatcttg taatcctgaa
361 ggctcctcag aaacagctct tcttcaaatc tatacattaa gacgactcga aatccacata
421 tcaaatatcc gagtgtagta aacattccaa aaccgtgatg gaatggaaca acacttaaaa
481 tcgcagtatc cggaatgatt tgattgccaa aaataggatc tctggcatgc gagaatctca
541 cgcaggcagt tctatgaggc agagcgacac ctttaggcag accagtagat ccagaggagt
601 tcatgatcag tgcaattgtc ttgtccctat cgaaggactc tggcacaaaa tcgtattcat
661 taaaaccggg aggtagatga gatgtgacga acgtgtacat cgactgaaat ccctggtaat
721 ccgttttaga atccatgata ataatttttt ggatgattgg gagctttttt tgcacgttca
781 aaattttttg caaccccttt ttggaaacga acaccacggt aggctgcgaa atgcccatac
841 tgttgagcaa ttcacgttca ttataaatgt cgttcgcggg cgcaactgca actccgataa
901 ataacgcgcc caacaccggc ataaagaatt gaagagagtt ttcactgcat acgacgattc
961 tgtgatttgt attcagccca tatcgtttca tagcttctgc caaccgaacg gacatttcga
1021 agtactcagc gtaagtgatg tccacctcga tatgtgcatc tgtaaaagca attgttccag
1081 gaaccagggc gtatctcttc atagccttat gcagttgctc tccagcggtt ccatcttcca
1141 gcggatagaa tggcgccggg cctttcttta tgtttttggc gtcttccatg gtggctttac
1201 caacagtacc ggaatgccaa gcttgggcag gagttgaggt tactgtgagt agtgattaaa
1261 gagagtgata gggaactctt gaacaagaga tgcaatttat actgttaatt ctggaaaaat
1321 attatggggg tgtcaaaatg tcctcgagga ggaaaaactg tttcatacag aaggcgtgga
1381 ggaaaaactg tttcatacag aaggcgtacg ccttctgtat gaaacagttt ttcctcctcg
1441 agggggggtg gggatacaaa agtaactcag aaaacatcgg ggctagcacg cgtaagagct
1501 cggtacctat cgatagagaa atgttctggc acctgcactt gcactgggga cagcctattt
1561 tgctagtttg ttttgtttcg ttttgttttg atggagagcg tatgttagta ctatcgattc
1621 acacaaaaaa ccaacacaca gatgtaatga aaataaagat attttattgc ggccgctcca
1681 agtacctccc gtaccttaat attacttact tatcatggta gcttgggctg gcgtaatagc
1741 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcaa
1801 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
1861 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
1921 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
1981 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
2041 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
2101 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
2161 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
2221 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
2281 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
2341 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
2401 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
2461 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
2521 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
2581 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
2641 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
2701 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
2761 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
2821 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
2881 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
2941 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
3001 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
3061 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
3121 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
3181 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
3241 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
3301 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
3361 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
3421 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
3481 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
3541 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
3601 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
3661 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
3721 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
3781 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
3841 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
3901 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
3961 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
4021 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
4081 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
4141 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg ctgccggcac
4201 ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga tagtcatgcc
4261 ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgacggat
4321 ccttatcgat tttaccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
4381 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
4441 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
4501 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgctcgaa
4561 gcggccggcc gccccgactc tagaattaca cggcgatctt tccgcccttc ttggccttta
4621 tgaggatctc tctgattttt cttgcgtcga gttttccggt aagacctttc ggtacttcgt
4681 ccacaaacac aactcctccg cgcaactttt tcgcggttgt tacttgactg gcgacgtaat
4741 ccacgatctc tttttccgtc atcgtctttc cgtgctccaa aacaacaacg gcggcgggaa
4801 gttcaccggc gtcatcgtcg ggaagacctg cgacacctgc gtcgaagatg ttggggtgtt
4861 ggagcaagat ggattccaat tcagcgggag ccacctgata gcctttgtac ttaatcagag
4921 acttcaggcg gtcaacgatg aagaagtgtt cgtcttcgtc ccagtaagct atgtctccag
4981 aatgtagcca tccatccttg tcaatcaagg cgttggtcgc ttccggattg tttacataac
5041 cggacataa
//
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