| 质粒名称: | 8xNFAT-ZsG-hCD8 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 153417 |
| 存储实验室: | Maki Nakayama |
|
载体骨架基本信息 |
|
| 载体名称: | pSIR |
| 载体出品公司: | Maki Nakayama |
| 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: | 哺乳动物表达，逆转录病毒 |
| 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: | 8x of NFAT binding motif follwed by ZsGreen-1 fluorescent protein gene |
| 物种来源: | Other |
| 插入大小: | 998 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | Gibson克隆 |
| 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: | 8xNFAT-ZsG-hCD8 was a gift from Maki Nakayama (Addgene plasmid # 153417 ; http://n2t.net/addgene:153417 ; RRID:Addgene_153417) |
| 参考文献部分: | Multiplex T Cell Stimulation Assay Utilizing a T Cell Activation Reporter-Based Detection System. Mann SE, Zhou Z, Landry LG, Anderson AM, Alkanani AK, Fischer J, Peakman M, Mallone R, Campbell K, Michels AW, Nakayama M. Front Immunol. 2020 Apr 9;11:633. doi: 10.3389/fimmu.2020.00633. eCollection 2020. 10.3389/fimmu.2020.00633 PubMed 32328071 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS 8xNFAT-ZsG-hCD8 7850 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION NFAT-driven ZsGreen-1 reporter gene with human CD8ab gene.
ACCESSION .
VERSION .
KEYWORDS 8xNFAT-ZsG-hCD8
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7850)
AUTHORS Mann SE, Zhou Z, Landry LG, Anderson AM, Alkanani AK, Fischer J,
Peakman M, Mallone R, Campbell K, Michels AW, Nakayama M
TITLE Multiplex T Cell Stimulation Assay Utilizing a T Cell Activation
Reporter-Based Detection System.
JOURNAL Front Immunol. 2020 Apr 9;11:633. doi: 10.3389/fimmu.2020.00633.
eCollection 2020.
PUBMED 32328071
REFERENCE 2 (bases 1 to 7850)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7850
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 585..760
/label=5' LTR (truncated)
/note="truncated long terminal repeat from Moloney murine
sarcoma virus"
misc_feature 823..1022
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 1223..1639
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1569..1591
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1607..1623
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
protein_bind 1673..1702
/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 1703..1732
/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 1733..1762
/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 1763..1792
/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 1793..1822
/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 1823..1852
/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 1853..1882
/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 1883..1912
/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)"
promoter 1924..1955
/label=minP
/note="minimal TATA-box promoter with low basal activity"
regulatory 1969..1978
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1975..2670
/codon_start=1
/product="Zoanthus green fluorescent protein"
/label=ZsGreen1
/note="mammalian codon-optimized"
/translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
AIASGSALP"
promoter 2693..3192
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(2728..2751)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 3125..3144
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
regulatory 3204..3213
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3924..3980
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
LTR 4793..5218
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
Moloney murine leukemia virus"
rep_origin complement(5388..5976)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5468..5487)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6147..7007)
/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 6770..6789
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7008..7112)
/gene="bla"
/label=AmpR promoter
primer_bind 7180..7198
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(7236..7258)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 7358..7377
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7571..7593
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 7721..7740
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcaataa aagagcccac
601 aacccctcac tcggcgcgcc agtcttccga tagactgcgt cgcccgggta cccgtattcc
661 caataaagcc tcttgctgtt tgcatccgaa tcgtggtctc gctgttcctt gggagggtct
721 cctctgagtg attgactacc cacgacgggg gtctttcatt tgggggctcg tccgggattt
781 ggagacccct gcccagggac caccgaccca ccaccgggag gtaagctggc cagcaactta
841 tctgtgtctg tccgattgtc tagtgtctat gtttgatgtt atgcgcctgc gtctgtacta
901 gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt tctgaacacc
961 cggccgcaac cctgggagac gtcccaggga ctttgggggc cgtttttgtg gcccgacctg
1021 aggaagggag tcgatgtgga atccgacccc gtcaggatat gtggttctgg taggagacga
1081 gaacctaaaa cagttcccgc ctccgtctga atttttgctt tcggtttgga accgaagccg
1141 cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt
1201 tctgtatttg tctgaaaatt agggccagac tgttaccact cccttaagtt tgaccttagg
1261 tcactggaaa gatgtcgagc ggatcgctca caaccagtcg gtagatgtca agaagagacg
1321 ttgggttacc ttctgctctg cagaatggcc aacctttaac gtcggatggc cgcgagacgg
1381 cacctttaac cgagacctca tcacccaggt taagatcaag gtcttttcac ctggcccgca
1441 tggacaccca gaccaggtcc cctacatcgt gacctgggaa gccttggctt ttgacccccc
1501 tccctgggtc aagccctttg tacaccctaa gcctccgcct cctcttcctc catccgcccc
1561 gtctctcccc cttgaacctc ctcgttcgac cccgcctcga tcctcccttt atccagccct
1621 cactccttct ctaggcgccg gaattgaaga tcatcatgtc tggatccaag ctggaggaaa
1681 aactgtttca tacagaaggc gtggaggaaa aactgtttca tacagaaggc gtggaggaaa
1741 aactgtttca tacagaaggc gtggaggaaa aactgtttca tacagaaggc gtggaggaaa
1801 aactgtttca tacagaaggc gtggaggaaa aactgtttca tacagaaggc gtggaggaaa
1861 aactgtttca tacagaaggc gtggaggaaa aactgtttca tacagaaggc gtcgcggaga
1921 ctctagaggg tatataatgg aagctcgaat tccagtcccc gagatctcgc caccatggcc
1981 cagtccaagc acggcctgac caaggagatg accatgaagt accgcatgga gggctgcgtg
2041 gacggccaca agttcgtgat caccggcgag ggcatcggct accccttcaa gggcaagcag
2101 gccatcaacc tgtgcgtggt ggagggcggc cccttgccct tcgccgagga catcttgtcc
2161 gccgccttca tgtacggcaa ccgcgtgttc accgagtacc cccaggacat cgtcgactac
2221 ttcaagaact cctgccccgc cggctacacc tgggaccgct ccttcctgtt cgaggacggc
2281 gccgtgtgca tctgcaacgc cgacatcacc gtgagcgtgg aggagaactg catgtaccac
2341 gagtccaagt tctacggcgt gaacttcccc gccgacggcc ccgtgatgaa gaagatgacc
2401 gacaactggg agccctcctg cgagaagatc atccccgtgc ccaagcaggg catcttgaag
2461 ggcgacgtga gcatgtacct gctgctgaag gacggtggcc gcttgcgctg ccagttcgac
2521 accgtgtaca aggccaagtc cgtgccccgc aagatgcccg actggcactt catccagcac
2581 aagctgaccc gcgaggaccg cagcgacgcc aagaaccaga agtggcacct gaccgagcac
2641 gccatcgcct ccggctccgc cttgccctga ctcgaggtta acgaattcta ccgggtaggg
2701 gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg
2761 cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg
2821 gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc
2881 ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact
2941 agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct ttggggcagc
3001 ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag gggtgggtcc
3061 gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc
3121 ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct
3181 ccgggccttt cgacctgcat cccgccacca tggccttacc agtgaccgcc ttgctcctgc
3241 cgctggcctt gctgctccac gccgccaggc cgagccagtt ccgggtgtcg ccgctggatc
3301 ggacctggaa cctgggcgag acagtggagc tgaagtgcca ggtgctgctg tccaacccga
3361 cgtcgggctg ctcgtggctc ttccagccgc gcggcgccgc cgccagtccc accttcctcc
3421 tatacctctc ccaaaacaag cccaaggcgg ccgaggggct ggacacccag cggttctcgg
3481 gcaagaggtt gggggacacc ttcgtcctca ccctgagcga cttccgccga gagaacgagg
3541 gctactattt ctgctcggcc ctgagcaact ccatcatgta cttcagccac ttcgtgccgg
3601 tcttcctgcc agcgaagccc accacgacgc cagcgccgcg accaccaaca ccggcgccca
3661 ccatcgcgtc gcagcccctg tccctgcgcc cagaggcgtg ccggccagcg gcggggggcg
3721 cagtgcacac gagggggctg gacttcgcct gtgatatcta catctgggcg cccttggccg
3781 ggacttgtgg ggtccttctc ctgtcactgg ttatcaccct ttactgcaac cacaggaacc
3841 gaagacgtgt ttgcaaatgt ccccggcctg tggtcaaatc gggagacaag cccagccttt
3901 cggcgagata cgtcggctcc ggagccacga acttctctct gttaaagcaa gcaggagacg
3961 tggaagaaaa ccccggtccc atgcggccgc ggctgtggct cctcctggcc gcgcagctga
4021 cagttctcca tggcaactca gtcctccagc agacccctgc atacataaag gtgcaaacca
4081 acaagatggt gatgctgtcc tgcgaagcta aaatctccct cagtaacatg cgcatctact
4141 ggctgagaca gcgccaggca ccgagcagtg acagtcacca cgagttcctg gccctctggg
4201 attccgcaaa agggactatc cacggtgaag aggtggaaca ggagaagata gctgtgtttc
4261 gggatgcaag ccggttcatt ctcaatctca caagcgtgaa gccggaagac agtggcatct
4321 acttctgcat gatcgtcggg agccccgagc tgaccttcgg gaagggaact cagctgagtg
4381 tggttgattt ccttcccacc actgcccagc ccaccaagaa gtccaccctc aagaagagag
4441 tgtgccggtt acccaggcca gagacccaga agggcccact ttgtagcccc atcacccttg
4501 gcctgctggt ggctggcatc ctggttctgc tggtttccct gggagtggcc atccacctgt
4561 gctgccggcg gaggagagcc cggcttcgtt tcatgaaaca gcctcaaggg gaaggtatat
4621 caggaacctt tgtcccccaa tgcctgcatg gatactacag caatactaca acctcacaga
4681 agctgcttaa cccatggatc ctgaaaacat agaagcttca ccagctgaag cctatagagt
4741 acgagccata gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag
4801 accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa
4861 atacataact gagaatagag aagttcagat caaggtcagg aacagatgga acagggtcga
4921 ccctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct
4981 tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag
5041 ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt gactgagtcg
5101 cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct
5161 gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatttg
5221 ggggctcgtc cgggatcggg agacccctgc ccagggacca ccgacccacc accgggaggt
5281 aagctggctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacat gtgagcaaaa
5341 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
5401 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
5461 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
5521 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
5581 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
5641 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
5701 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
5761 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
5821 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
5881 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
5941 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
6001 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
6061 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
6121 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
6181 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
6241 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
6301 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
6361 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
6421 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
6481 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
6541 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
6601 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
6661 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
6721 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
6781 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
6841 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
6901 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
6961 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
7021 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
7081 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
7141 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
7201 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
7261 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
7321 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
7381 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
7441 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
7501 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
7561 ccagggtttt cccagtcacg acgttgtaaa acgacggcgc aaggaatggt gcatgcaagg
7621 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
7681 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
7741 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagaggc
7801 gattagtcca atttgttaaa gacaggatat cccaacatta ccgccatgtt
//
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