| 质粒名称: | pirespuro2-CTLA4-mEos2 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 98285 |
| 存储实验室: | Mike Heilemann |
|
载体骨架基本信息 |
|
| 载体名称: | pIRESpuro2 |
| 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: | 哺乳细胞表达 |
| 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: | CTLA4 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | mEos2（插入件上的C端子） |
| 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 | CMV-F |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pirespuro2-CTLA4-mEos2 was a gift from Mike Heilemann (Addgene plasmid # 98285 ; http://n2t.net/addgene:98285 ; RRID:Addgene_98285) |
| 参考文献部分: | One, two or three? Probing the stoichiometry of membrane proteins by single-molecule localization microscopy. Fricke F, Beaudouin J, Eils R, Heilemann M. Sci Rep. 2015 Sep 11;5:14072. doi: 10.1038/srep14072. 10.1038/srep14072 PubMed 26358640 |
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 pirespuro2-CTLA4-mEos2 6485 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION mammalian expression of CTLA4-mEos2.
ACCESSION .
VERSION .
KEYWORDS pirespuro2-CTLA4-mEos2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6485)
AUTHORS Fricke F, Beaudouin J, Eils R, Heilemann M
TITLE One, two or three? Probing the stoichiometry of membrane proteins by
single-molecule localization microscopy.
JOURNAL Sci Rep. 2015 Sep 11;5:14072. doi: 10.1038/srep14072.
PUBMED 26358640
REFERENCE 2 (bases 1 to 6485)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6485
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 11..390
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 391..594
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 545..565
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 639..658
/label=T7
/note="T7 promoter, forward primer"
promoter 639..657
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 709..718
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
intron 2128..2357
/label=chimeric intron
/note="chimera between introns from adenovirus and
immunoglobulin heavy chain genes"
primer_bind 2340..2359
/label=pMT2-F
/note="Synthetic intron, forward primer"
misc_feature 2518..2980
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(2574..2591)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 2801..2820
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 3013..3612
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(3013..3032)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3509..3529
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
primer_bind complement(3748..3765)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3754..3978
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind 4025..4044
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4104..4120)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4104..4120)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4117..4139)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4128..4144
/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)."
promoter complement(4152..4182)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4197..4218
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4335..4352)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4506..5094)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4586..4605)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5265..6125)
/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 5888..5907
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6126..6230)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6305..6324)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
61 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
121 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
181 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
241 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
301 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
361 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
421 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
481 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
541 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
601 ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag
661 acccaagctt ggtaccgagc tcggatcgat atctgcggcc tagctagcgc caccatggct
721 tgccttggat ttcagcggca caaggctcag ctgaacctgg ctaccaggac ctggccctgc
781 actctcctgt tttttcttct cttcatccct gtcttctgca aagcaatgca cgtggcccag
841 cctgctgtgg tactggccag cagccgaggc atcgccagct ttgtgtgtga gtatgcatct
901 ccaggcaaag ccactgaggt ccgggtgaca gtgcttcggc aggctgacag ccaggtgact
961 gaagtctgtg cggcaaccta catgatgggg aatgagttga ccttcctaga tgattccatc
1021 tgcacgggca cctccagtgg aaatcaagtg aacctcacta tccaaggact gagggccatg
1081 gacacgggac tctacatctg caaggtggag ctcatgtacc caccgccata ctacctgggc
1141 ataggcaacg gaacccagat ttatgtaatt gatccagaac cgtgcccaga ttctgacttc
1201 ctcctctgga tccttgcagc agttagttcg gggttgtttt tttatagctt tctcctcaca
1261 gctgtttctt tgagcaaaat gctaaagaaa agaagccctc ttacaacagg ggtcggaggt
1321 ggaccggtgc cccagtggga gggattcgct gcgctgctgg caaccccggt cgctactatg
1381 tctgctatca agccagacat gaagatcaag ttgagaatgg aaggtaacgt taacggtcac
1441 cacttcgtta tcgacggtga cggtactggt aagccattcg aaggtaagca atctatggac
1501 ttggaagtta aggaaggtgg tccattgcca ttcgctttcg acatcttgac tactgctttc
1561 cactacggta acagagtttt cgctaagtac ccagacaaca tccaagacta cttcaagcaa
1621 tctttcccaa agggttactc ttgggaaaga tctttgactt tcgaagacgg tggtatctgt
1681 atcgctagaa acgacatcac tatggaaggt gacactttct acaacaaggt tagattctac
1741 ggtactaact tcccagctaa cggtccagtt atgcaaaaga agactttgaa gtgggaacca
1801 tctactgaaa agatgtacgt tagagacggt gttttgactg gtgacatcca catggctttg
1861 ttgttggaag gtaacgctca ctacagatgt gacttcagaa ctacttacaa ggctaaggaa
1921 aagggtgtta agttgccagg ttaccacttc gttgaccact gtatcgaaat cttgtctcac
1981 gacaaggact acaacaaggt taagttgtac gaacacgctg ttgctcactc tggtttgcca
2041 gacaacgcta gaagataaag cggccgcata gataactgat ccagtgtgct ggaattaatt
2101 cgctgtctgc gagggccagc tgttggggtg agtactccct ctcaaaagcg ggcatgactt
2161 ctgcgctaag attgtcagtt tccaaaaacg aggaggattt gatattcacc tggcccgcgg
2221 tgatgccttt gagggtggcc gcgtccatct ggtcagaaaa gacaatcttt ttgttgtcaa
2281 gcttgaggtg tggcaggctt gagatctggc catacacttg agtgacaatg acatccactt
2341 tgcctttctc tccacaggtg tccactccca ggtccaactg caggtcgagc atgcatctag
2401 ggcggccaat tccgcccctc nnncccctaa cgttactggc cgaagccgct tggaataagg
2461 ccggtgtgcg tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag
2521 ggcccggaaa cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc
2581 caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg
2641 aagacaaaca acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag
2701 gtgcctctgc ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca
2761 gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt
2821 caacaagggg ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc
2881 tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa
2941 ccacggggac gtggttttcc tttgaaaaac acgatgataa gcttgccaca acccacaagg
3001 agacgacctt ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
3061 ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
3121 gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
3181 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
3241 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
3301 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
3361 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
3421 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
3481 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
3541 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
3601 cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc
3661 catggctccg accgaagcca cccggggcgg ccccgccgac cccgcacccg cccccgaggc
3721 ccaccgactc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
3781 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
3841 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
3901 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
3961 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctcg agtgcattct
4021 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
4081 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
4141 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
4201 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
4261 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
4321 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
4381 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
4441 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
4501 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
4561 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
4621 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
4681 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
4741 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
4801 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
4861 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
4921 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
4981 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
5041 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
5101 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
5161 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
5221 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
5281 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
5341 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
5401 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
5461 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
5521 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
5581 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
5641 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
5701 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
5761 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
5821 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
5881 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
5941 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
6001 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
6061 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
6121 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
6181 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
6241 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
6301 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
6361 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
6421 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
6481 ccaga
//
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