| 质粒名称: | pDisplay-AP-CFP-TM |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 20861 |
| 存储实验室: | Alice Ting |
|
载体骨架基本信息 |
|
| 载体名称: | pDisplay |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5300 bp |
| 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: | AP-CFP-TM |
| 物种来源: | E. Coli |
| 插入大小: | 1092 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: |
HA（主干上的N终端）
CFP（插入件上的C端子）
myc（主干上的C终端）
|
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BglII(未破坏) |
| 3' 克隆位点: | SacII(未破坏) |
| 5' 测序引物 | T7 |
| 3' 测序引物 | T7 term |
|
文献引用方法 |
|
| 材料和方法部分: | pDisplay-AP-CFP-TM was a gift from Alice Ting (Addgene plasmid # 20861 ; http://n2t.net/addgene:20861 ; RRID:Addgene_20861) |
| 参考文献部分: | Targeting quantum dots to surface proteins in living cells with biotin ligase. Howarth M, Takao K, Hayashi Y, Ting AY. Proc Natl Acad Sci U S A. 2005 May 24. 102(21):7583-8. 10.1073/pnas.0503125102 PubMed 15897449 |
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 pDisplay-AP-CFP-TM 6097 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pDisplay-AP-CFP-TM
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6097)
AUTHORS Howarth M, Takao K, Hayashi Y, Ting AY
TITLE Targeting quantum dots to surface proteins in living cells with
biotin ligase.
JOURNAL Proc Natl Acad Sci U S A. 2005 May 24. 102(21):7583-8.
PUBMED 15897449
REFERENCE 2 (bases 1 to 6097)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6097
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 10..389
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 390..593
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 544..564
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 638..657
/label=T7
/note="T7 promoter, forward primer"
promoter 638..656
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
sig_peptide 737..799
/product="leader sequence from mouse immunoglobulin kappa
light chain"
/label=Ig-kappa leader
CDS 800..826
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 854..898
/codon_start=1
/product="peptide tag that allows for enzymatic
biotinylation"
/label=AviTag(TM)
/translation="GLNDIFEAQKIEWHE"
CDS 908..1624
/codon_start=1
/product="enhanced CFP"
/label=ECFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(953..974)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1214..1233)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1561..1582
/label=EGFP-C
/note="EGFP, forward primer"
CDS 1646..1675
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 1679..1825
/codon_start=1
/product="transmembrane domain from platelet derived growth
factor receptor beta"
/label=PDGFR-beta TM domain
/translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
KKPR"
primer_bind complement(1847..1864)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1853..2077
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin complement(2209..2797)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2289..2308)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
polyA_signal 3126..3173
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
primer_bind 3198..3217
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
CDS complement(3405..4193)
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDQEHQGL
APAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIALA
TRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3516..3535)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 4120..4139
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
promoter complement(4228..4588)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4242..4380
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(4299..4318)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 4545..4565
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
CDS complement(4648..5508)
/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 5271..5290
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5509..5613)
/gene="bla"
/label=AmpR promoter
rep_origin 5640..6095
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5727..5746)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5937..5958
/label=F1ori-F
/note="F1 origin, forward primer"
ORIGIN
1 gcgcgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
181 caatagggac tttccattga cgtcaatggg tggactattt acggtaaact gcccacttgg
241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
601 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
661 cccaagcttg gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattcgg
721 cttggggata tccaccatgg agacagacac actcctgcta tgggtactgc tgctctgggt
781 tccaggttcc actggtgact atccatatga tgttccagat tatgctgggg cccagccggc
841 cagatctggc ggcggcctga acgacatctt cgaggcccag aagatcgagt ggcacgaggg
901 cgcgccgatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
961 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
1021 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
1081 gcccaccctc gtgaccaccc tgacctgggg cgtgcagtgc ttcagccgct accccgacca
1141 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
1201 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
1261 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
1321 ggggcacaag ctggagtaca actacatcag ccacaacgtc tatatcaccg ccgacaagca
1381 gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg gcagcgtgca
1441 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
1501 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
1561 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
1621 caagcggccg cggctgcagg tcgacgaaca aaaactcatc tcagaagagg atctgaatgc
1681 tgtgggccag gacacgcagg aggtcatcgt ggtgccacac tccttgccct ttaaggtggt
1741 ggtgatctca gccatcctgg ccctggtggt gctcaccatc atctccctta tcatcctcat
1801 catgctttgg cagaagaagc cacgttaggc ggccgctcga gatcagcctc gactgtgcct
1861 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
1921 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
1981 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
2041 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagt
2101 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
2161 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
2221 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
2281 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
2341 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
2401 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
2461 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
2521 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
2581 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
2641 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
2701 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
2761 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
2821 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
2881 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
2941 tatatatgag taacctgagg ctatggcagg gcctgccgcc ccgacgttgg ctgcgagccc
3001 tgggccttca cccgaacttg gggggtgggg tggggaaaag gaagaaacgc gggcgtattg
3061 gccccaatgg ggtctcggtg gggtatcgac agagtgccag ccctgggacc gaaccccgcg
3121 tttatgaaca aacgacccaa caccgtgcgt tttattctgt ctttttattg ccgtcatagc
3181 gcgggttcct tccggtattg tctccttccg tgtttcagtt agcctccccc tagggtgggc
3241 gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac
3301 gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtgatggcag
3361 gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca
3421 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg
3481 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg
3541 tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
3601 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
3661 tcgggcatgc tcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
3721 tgatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt
3781 ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca
3841 tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc
3901 ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct
3961 gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca
4021 ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc
4081 cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc
4141 ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac
4201 gatcctcatc ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc
4261 tcactacttc tggaatagct cagaggccga ggaggcggcc tcggcctctg cataaataaa
4321 aaaaattagt cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat
4381 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact
4441 tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat
4501 gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac
4561 attccacagc tggttctttc cgcctcagga ctcttccttt ttcaataaat caatctaaag
4621 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
4681 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
4741 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
4801 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
4861 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
4921 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
4981 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
5041 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
5101 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
5161 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
5221 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
5281 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
5341 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
5401 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
5461 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
5521 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
5581 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
5641 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
5701 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
5761 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
5821 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
5881 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
5941 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
6001 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
6061 cgcgaatttt aacaaaatat taacgcttac aatttac
//
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