| 质粒名称: | pEF6a-FKBP-CD28 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 113401 |
| 存储实验室: | Min Zhuang |
|
载体骨架基本信息 |
|
| 载体名称: | pEF6a |
| 完整质粒大小: | 6782bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | FKBP-CD28 |
| 物种来源: | 未知 |
| 插入大小: | 1002 bp |
| 启动子: | EF-1α |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | EF-1α |
|
文献引用方法 |
|
| 材料和方法部分: | pEF6a-FKBP-CD28 was a gift from Min Zhuang (Addgene plasmid # 113401 ; http://n2t.net/addgene:113401 ; RRID:Addgene_113401) |
| 参考文献部分: | A proximity-tagging system to identify membrane protein-protein interactions. Liu Q, Zheng J, Sun W, Huo Y, Zhang L, Hao P, Wang H, Zhuang M. Nat Methods. 2018 Aug 13. pii: 10.1038/s41592-018-0100-5. doi: 10.1038/s41592-018-0100-5. 10.1038/s41592-018-0100-5 PubMed 30104635 |
买家导航
pEF6a-FKBP-CD28载体质粒基本信息
pEF6a-FKBP-CD28质粒图谱载体图谱和pEF6a-FKBP-CD28载体序列质粒序列多克隆位点信息
pEF6a-FKBP-CD28质粒载体简介
pEF6a-FKBP-CD28质粒序列载体序列
LOCUS pEF6a-FKBP-CD28 6782 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses FKBP-CD28 in mammalian cell.
ACCESSION .
VERSION .
KEYWORDS pEF6a-FKBP-CD28
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6782)
AUTHORS Liu Q, Zheng J, Sun W, Huo Y, Zhang L, Hao P, Wang H, Zhuang M
TITLE A proximity-tagging system to identify membrane protein-protein
interactions.
JOURNAL Nat Methods. 2018 Aug 13. pii: 10.1038/s41592-018-0100-5. doi:
10.1038/s41592-018-0100-5.
PUBMED 30104635
REFERENCE 2 (bases 1 to 6782)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6782
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..1179
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 232..1170
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1127..1147
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
primer_bind 1196..1215
/label=T7
/note="T7 promoter, forward primer"
promoter 1196..1214
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1323..1643
/codon_start=1
/product="human FK506-binding protein FKBP12"
/label=FKBP
/translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
KLE"
CDS 2314..2343
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 2359..2376
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(2399..2416)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2405..2629
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2675..3103
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2762..2781)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2972..2993
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3112..3132)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3117..3446
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3297..3432
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3359..3378
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 3494..3541
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 3560..3958
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
polyA_signal 4116..4237
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4153..4172)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4207..4226
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4286..4302)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4286..4302)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4299..4321)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4310..4326
/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(4334..4364)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4379..4400
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4517..4534)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4677..5265)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4757..4776)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5436..6296)
/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 6059..6078
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6297..6401)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6476..6495)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtactaata
1201 cgactcacta tagggagacc caagctggct aggtaagctt ggtaccgagc tcggccacca
1261 tgctcaggct gctcttggct ctcaacttat tcccttcaat tcaagtaaca ggaggcggaa
1321 gcggagtgca ggtagaaaca atctccccgg gagatggccg cacgttcccc aagaggggac
1381 agacctgtgt ggtgcactac accggtatgc tcgaagacgg caagaagttc gatagctccc
1441 gagaccgaaa caagcccttc aagttcatgc tgggcaagca agaggtcata cgcggttggg
1501 aagaaggcgt ggcccagatg agcgtagggc agcgcgccaa gctgaccatt agccccgact
1561 acgcctacgg ggccaccggg caccccggca tcattccacc ccatgcgaca ctcgtctttg
1621 atgtggagct gctcaagctg gaaggaggca gcggcaacaa gattttggtg aagcagtcgc
1681 ccatgcttgt agcgtacgac aatgcggtca accttagctg caagtattcc tacaatctct
1741 tctcaaggga gttccgggca tcccttcaca aaggactgga tagtgctgtg gaagtctgtg
1801 ttgtatatgg gaattactcc cagcagcttc aggtttactc aaaaacgggg ttcaactgtg
1861 atgggaaatt gggcaatgaa tcagtgacat tctacctcca gaatttgtat gttaaccaaa
1921 cagatattta cttctgcaaa attgaagtta tgtatcctcc tccttaccta gacaatgaga
1981 agagcaatgg aaccattatc catgtgaaag ggaaacacct ttgtccaagt cccctatttc
2041 ccggaccttc taagcccttt tgggtgctgg tggtggttgg tggagtcctg gcttgctata
2101 gcttgctagt aacagtggcc tttattattt tctgggtgag gagtaagagg agcaggctcc
2161 tgcacagtga ctacatgaac atgactcccc gccgccccgg gcccacccgc aagcattacc
2221 agccctatgc cccaccacgc gacttcgcag cctatcgctc ctagctgcag atatccagca
2281 cagtggcggc cgctcgagtc tagagggccc ttcgaacaaa aactcatctc agaagaggat
2341 ctgaatatgc ataccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc
2401 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
2461 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
2521 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
2581 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
2641 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
2701 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
2761 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
2821 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
2881 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
2941 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
3001 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
3061 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
3121 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
3181 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
3241 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
3301 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
3361 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
3421 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
3481 ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
3541 caaggtgagg aactaaacca tggccaagcc tttgtctcaa gaagaatcca ccctcattga
3601 aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc
3661 agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg
3721 accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac
3781 ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg
3841 acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca
3901 gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaagc
3961 acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct
4021 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
4081 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
4141 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
4201 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
4261 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
4321 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
4381 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
4441 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
4501 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
4561 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
4621 gaaagaacat gtgagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
4681 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
4741 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
4801 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
4861 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
4921 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
4981 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
5041 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
5101 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
5161 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
5221 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
5281 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
5341 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
5401 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
5461 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
5521 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
5581 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
5641 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
5701 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
5761 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
5821 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
5881 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
5941 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
6001 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
6061 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
6121 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
6181 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
6241 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
6301 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
6361 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
6421 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
6481 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
6541 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
6601 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
6661 cgcgttgaca ttgattattg actaggcttt tgcaaaaagc tttgcaaaga tggataaagt
6721 tttaaacaga gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa
6781 tt
//
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