| 质粒名称: | pCE-BiFC-VC155 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 22020 |
| 存储实验室: | Vladislav Verkhusha |
|
载体骨架基本信息 |
|
| 载体名称: | pPD4983 |
| 载体出品公司: | Fire lab 1995 kit |
| 载体抗性: | 氨苄霉素 |
| 空载体大小: | 3969 bp |
| 完整质粒大小: | 4224 bp |
| 载体类型: | Worm Expression |
| 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: | VC155 |
| 物种来源: | Aequorea Victoria |
| 插入大小: | 255 bp |
| 基因又名: | Venus(155-238, A206K) |
| 基因突变: | Changed Alanine 206 to Lysine |
| 标签/融合蛋白: | HA (N terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | KpnI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | n/a |
| 3' 测序引物 | n/a |
|
文献引用方法 |
|
| 材料和方法部分: | pCE-BiFC-VC155 was a gift from Chang-Deng Hu (爱迪基因 plasmid # 22020) |
| 参考文献部分: | Visualization of protein interactions in living Caenorhabditis elegans using bimolecular fluorescence complementation analysis. Shyu YJ, Hiatt SM, Duren HM, Ellis RE, Kerppola TK, Hu CD. Nat Protoc. 2008 . 3(4):588-96. 10.1038/nprot.2008.16 PubMed 18388940 |
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 pCE-BiFC-VC155 4224 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCE-BiFC-VC155
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4224)
AUTHORS Shyu YJ, Hiatt SM, Duren HM, Ellis RE, Kerppola TK, Hu CD
TITLE Visualization of protein interactions in living Caenorhabditis
elegans using bimolecular fluorescence complementation analysis.
JOURNAL Nat Protoc. 2008 . 3(4):588-96.
PUBMED 18388940
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pCE-BiFC-VC155|
FEATURES Location/Qualifiers
source 1..4224
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 4..30
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 112..363
/codon_start=1
/product="C-terminal fragment of mVenus for use in
bimolecular fluorescence complementation (BiFC) (Kodama and
Hu, 2010)"
/label=VC155
/translation="DKQKNGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNH
YLSYQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
primer_bind 300..321
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(1231..1253)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 1353..1372
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(1450..1468)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1536..1640
/gene="bla"
/label=AmpR promoter
CDS 1641..2501
/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 complement(1859..1878)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2672..3260
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3161..3180
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3414..3431
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 3548..3569
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3584..3614
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3622..3638
/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)."
primer_bind 3627..3649
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 3646..3662
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3646..3662
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
ORIGIN
1 atgtacccat acgatgttcc agattacgct cttatggcca tggaggcccg aattcggtcg
61 accgagatct ctcgaggtac cggaggtggc gggagcggag gtggcgggag tgacaagcag
121 aagaacggca tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag
181 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
241 aaccactacc tgagctacca gtccaaactg agcaaagacc ccaacgagaa gcgcgatcac
301 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
361 aagtaagcgg ccgcaggatc cagacatgat aagatacgat atctgagctc cgcatcggcc
421 gctgtcatca gatcgccatc tcgcgcccgt gcctctgact tctaagtcca attactcttc
481 aacatcccta catgctcttt ctccctgtgc tcccaccccc tatttttgtt attatcaaaa
541 aaacttcttc ttaatttctt tgttttttag cttcttttaa gtcacctcta acaatgaaat
601 tgtgtagatt caaaaataga attaattcgt aataaaaagt cgaaaaaaat tgtgctccct
661 ccccccatta ataataattc tatcccaaaa tctacacaat gttctgtgta cacttcttat
721 gtttttttta cttctgataa attttttttg aaacatcata gaaaaaaccg cacacaaaat
781 accttatcat atgttacgtt tcagtttatg accgcaattt ttatttcttc gcacgtctgg
841 gcctctcatg acgtcaaatc atgctcatcg tgaaaaagtt ttggagtatt tttggaattt
901 ttcaatcaag tgaaagttta tgaaattaat tttcctgctt ttgctttttg ggggtttccc
961 ctattgtttg tcaagagttt cgaggacggc gtttttcttg ctaaaatcac aagtattgat
1021 gagcacgatg caagaaagat cggaagaagg tttgggtttg aggctcagtg gaaggtgagt
1081 agaagttgat aatttgaaag tggagtagtg tctatggggt ttttgcctta aatgacagaa
1141 tacattccca atataccaaa cataactgtt tcctactagt cggccgtacg ggccctttcg
1201 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
1261 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
1321 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
1381 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
1441 gccttaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
1501 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
1561 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
1621 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
1681 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
1741 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
1801 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
1861 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
1921 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
1981 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
2041 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
2101 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
2161 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
2221 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
2281 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
2341 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
2401 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
2461 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
2521 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
2581 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
2641 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
2701 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
2761 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
2821 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
2881 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
2941 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
3001 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
3061 agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
3121 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
3181 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
3241 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
3301 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
3361 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
3421 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
3481 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
3541 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
3601 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
3661 accatgatta cgccaagctt gcatgcctgc aggtcgactc tagaggatca ccaaaaacgg
3721 aacgttgagc tggacggaaa tagtggtaaa gtgacatgat tatagtttga agatttctaa
3781 tttcacaatt agagcaaatg ttgttcggta tttattttca acggtattta tactattttc
3841 cacctttttc tagaacattc gagctgcttg ttgcaaaagg agggcgactc acattcggta
3901 catggaaaag tagtgtacac aataaagaga cccagataca ttttccgtct gcgtctcttt
3961 gcacccaccg ggagtatttt caaacgaatg catctaggac cttctagaac attctgtaag
4021 gctgcagaat gcgggtatat aaggaaagcg ggctcagagg aagccaacac gctttgttct
4081 agtgcatcta aaaaacttcg aaaatcctca tcgggatccc cgggattggc caaaggaccc
4141 aaaggtatgt ttcgaatgat actaacataa catagaacat tttcaggagg acccttggct
4201 agcaattgta cccgcgggcc cacc
//
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