| 质粒名称: | Kif5A-GFP-CIBN |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 102252 |
| 存储实验室: | Bianxiao Cui |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-N1 |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 3800 bp |
| 完整质粒大小: | 6800 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Kif5A-GFP-CIB1,Kif5A,CIB1,CIBN |
| 物种来源: | R. norvegicus (rat), A. thaliana (mustard weed) |
| 插入大小: | 3000 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NheI(未破坏) |
| 3' 克隆位点: | SalI(未破坏) |
| 5' 测序引物 | CMV forward |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | Kif5A-GFP-CIBN was a gift from Bianxiao Cui (Addgene plasmid # 102252 ; http://n2t.net/addgene:102252 ; RRID:Addgene_102252) |
| 参考文献部分: | Optogenetic control of molecular motors and organelle distributions in cells. Duan L, Che D, Zhang K, Ong Q, Guo S, Cui B. Chem Biol. 2015 May 21;22(5):671-82. doi: 10.1016/j.chembiol.2015.04.014. Epub 2015 May 9. 10.1016/j.chembiol.2015.04.014 PubMed 25963241 |
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 Kif5A-GFP-CIBN 6796 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses fusion of kinesin heavy chain 5A (1-572) with GFP and CIB1
(1-170).
ACCESSION .
VERSION .
KEYWORDS Kif5A-GFP-CIBN
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6796)
AUTHORS Duan L, Che D, Zhang K, Ong Q, Guo S, Cui B
TITLE Optogenetic control of molecular motors and organelle distributions
in cells.
JOURNAL Chem Biol. 2015 May 21;22(5):671-82. doi:
10.1016/j.chembiol.2015.04.014. Epub 2015 May 9.
PUBMED 25963241
REFERENCE 2 (bases 1 to 6796)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6796
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 61..364
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 519..539
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 606..615
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2334..3050
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 2334..3050
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(2379..2400)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2640..2659)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2987..3008
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 3584..3705
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3621..3640)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3675..3694
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(3712..4167)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3849..3870)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4061..4080
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4194..4298
/gene="bla"
/label=AmpR promoter
promoter 4300..4657
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(4323..4343)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 4508..4643
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4570..4589
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4692..5486
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(4746..4765)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5356..5375
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(5674..5693)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5718..5765
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 6094..6682
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6583..6602
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
601 agcttgccac catgggcgcg cctgaattcg cggagaccaa taacgaatgc agcatcaagg
661 tgctttgccg atttcggccc ctgaaccagg ccgagatcct gcggggggac aagttcatcc
721 ccattttcca aggggacgac agcgtcatta ttgggggaaa gccatatgtc ttcgaccgtg
781 tattcccccc aaacacgact caggagcagg tttaccacgc ctgtgccatg cagatcgtca
841 aagacgtcct tgctggctac aacggcacaa tttttgctta tggacagaca tcctcaggga
901 aaacgcacac catggaggga aagctgcacg accctcagct catgggcatc attccccgga
961 tcgcccgaga catcttcaac cacatctact ccatggatga gaaccttgag ttccacatta
1021 aggtgtctta ctttgagatt tacctggata agatccgtga ccttttggat gtgaccaaga
1081 caaacctgtc tgtgcatgag gacaaaaacc gggtgccgtt tgtcaagggt tgtaccgaac
1141 gctttgtgtc cagcccagag gagattctgg atgtgattga tgaggggaag tcgaaccgtc
1201 acgttgccgt caccaacatg aatgagcaca gctctcggag tcacagcatc ttcctcatca
1261 acatcaagca ggagaacata gagacggagc agaagctcag cgggaaactg taccttgtgg
1321 atctggccgg aagcgagaag gtcagcaaga caggagcaga gggagccgtt ctggatgagg
1381 caaagaatat caataagtcg ctgtcggccc tggggaatgt gatctctgct ctggccgagg
1441 gcactaaaag ctacgtgccg taccgtgaca gcaaaatgac gaggattctc caggactctc
1501 tgggagggaa ctgcaggacc accatgttca tctgctgctc accatccagc tacaatgacg
1561 cagagaccaa gtccacactc atgtttgggc agcgggcaaa gaccatcaag aacactgcct
1621 cagtgaatct ggagttgact gctgagcaat ggaagaagaa atatgagaag gagaaggaga
1681 agaccaaggc ccagaaggag acaattgcga agctggaggc tgagcttagc cggtggcgca
1741 atggagagaa tgtgcctgag actgagcgcc tggctggaga ggactcagct ctggcagccg
1801 agatctgtga ggagacccct gtgaatgaca actcatccat cgtggtacgc atcgcaccgg
1861 aggaaaggca gaaatacgag gaagagatcc gccgtctcta caagcagctc gacgacaagg
1921 atgatgagat caaccagcag agccaactca ttgagaagct gaagcagcag atgctggacc
1981 aggaagagct gctagtgtcc actcggggag acaatgagaa ggtccagcgg gagctgagcc
2041 acctgcagtc tgagaacgat gccgcgaagg aggaggtgaa ggaagtgttg caggccctgg
2101 aggagctggc agtcaactat gaccagaagt cccaggaggt ggaggagaag agccagcaga
2161 accagctgct ggtggatgag ctgtcccaga aagtggccac catgctgtcc ctggagtctg
2221 agctacagcg gctccaggaa gtcagtggac accagcgaaa acggattgct gaggtgctga
2281 atggactgat gaaggacctg agtgagttca gtgtcatcgt gggcaagtcg actatggtga
2341 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
2401 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
2461 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
2521 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
2581 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
2641 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
2701 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
2761 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
2821 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
2881 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
2941 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
3001 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag ggatccatga
3061 atggagctat aggaggtgac cttttgctca attttcctga catgtcggtc ctagagcgcc
3121 aaagggctca cctcaagtac ctcaatccca cctttgattc tcctctcgcc ggcttctttg
3181 ccgattcttc aatgattacc ggcggcgaga tggacagcta tctttcgact gccggtttga
3241 atcttccgat gatgtacggt gagacgacgg tggaaggtga ttcaagactc tcaatttcgc
3301 cggaaacgac gcttgggact ggaaatttca aggcagcgaa gtttgataca gagactaagg
3361 attgtaatga ggcggcgaag aagatgacga tgaacagaga tgacctagta gaagaaggag
3421 aagaagagaa gtcgaaaata acagagcaaa acaatgggag cacaaaaagc atcaagaaga
3481 tgaaacacaa agccaagaaa gaagagaaca atttctctaa tgattcatct aaagtgacga
3541 aggaattgga gaaaacggat tatatttaac aattgttgtt gttaacttgt ttattgcagc
3601 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
3661 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa
3721 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
3781 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
3841 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
3901 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
3961 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
4021 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
4081 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
4141 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
4201 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
4261 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
4321 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
4381 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
4441 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
4501 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
4561 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
4621 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
4681 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
4741 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
4801 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
4861 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
4921 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
4981 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
5041 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
5101 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
5161 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
5221 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
5281 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
5341 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
5401 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
5461 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
5521 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
5581 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
5641 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
5701 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
5761 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
5821 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
5881 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
5941 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
6001 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
6061 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
6121 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
6181 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
6241 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
6301 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
6361 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
6421 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
6481 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
6541 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
6601 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
6661 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
6721 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
6781 tattaccgcc atgcat
//
