pEGFP-Notch1-NICD-r

Catalog No. PVT10478
Packing 2ug
 

pEGFP-Notch1-NICD-r Information

Promoter: CMV promoter

Replicator: pUC ori, F1 ori, SV40 ori

Terminator: SV40 poly (A) signal

Plasmid classification: gene library plasmid; rat gene plasmid; rat eukaryotic plasmid.

Plasmid size: 7079bp

Plasmid tagging: C-EGFP

Prokaryotic resistance: Kan (50 g/ml)

Selection marker: Neo/G418

Cloning strains: E. coli DH5 and E.

Culture conditions: 37 C, LB, aerobic.

Expression host: mammalian cells such as 293T

Culture conditions: 5%CO2, 37 C

Induction mode: no need to induce

5'sequencing primers: pEGFP-N-5 (TGGGAGGTCTATATAAGCAGAG)

3'sequencing primers: pEGFP-N-3 (CGTCGCCGTCCAGCTCGACCAG)

pEGFP-Notch1-NICD-r Description
pEGFP-Notch1-NICD-r expresses EGFP-Notch1-NICD fusion protein under the action of CMV strong promoter. The transmembrane macromolecule Notch1 receptor is mainly involved in the selection of cell fate during development. The active fragment-cytoplasmic domain (NICD) of G1743 and V1744 can be transported into the nucleus and activate the expression of target genes. Notch1 plays an important role in cell proliferation, differentiation, programmed death, morphogenesis and organogenesis during development. Cheng. Our constructed pEGFP-Notch1-NICD-r plasmid was transfected into COS-7L cells and green fluorescent protein was observed as nuclear localization, suggesting that the cloned NICD gene is in line with the further study of the function of NICD in post-division neurons.

pEGFP-Notch1-NICD-r  Multiple cloning site

pEGFP-Notch1-NICD-r Sequence

LOCUS       Exported                7097 bp ds-DNA     circular SYN 04-JUL-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pEGFP-Notch1-NICD-r
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7097)
  AUTHORS   yinchu
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, July 4, 2017 from SnapGene Viewer 3.3.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7097
                     /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"
     misc_feature    637..3002
                     /label=Notch1-NICD-r
                     /note="Rattus norvegicus notch 1 (Notch1), mRNA
                    
                     NCBI Reference Sequence: NM_001105721.1
                    
                     "
     CDS             3043..3762
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    3885..4006
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4013..4468)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4495..4599
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4601..4958
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4809..4944
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4993..5787
                     /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"
     polyA_signal    6019..6066
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6395..6983
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
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 ccggactcag atctcgagct caagcttcga attctgatgc tgctgtcccg caagcgcagg
      661 cggcagcatg gccagctctg gttccctgag ggtttcaaag tgtcagaggc cagcaagaag
      721 aagcggagag aacccctcgg cgaggactca gtcggcctca agcccctgaa gaacgcctca
      781 gatggtgccc tgatggacga caatcagaac gagtgggggg acgaagacct ggagaccaag
      841 aagttccggt ttgaggaacc agtggttctc cctgacctgg atgatcagac tgaccaccgg
      901 cagtggaccc agcagcacct ggatgccgct gacctacgtg tgtctgccat ggccccaacg
      961 ccgcctcagg gggaggtaga tgctgactgc atggacgtca atgttcgagg accagatggc
     1021 ttcactcccc taatgattgc ctcctgcagc ggagggggcc tggagacagg caacagtgag
     1081 gaagaagaag atgcacctgc tgtcatctcc gacttcatct atcagggtgc cagcttgcac
     1141 aaccagacgg accgcacagg ggagactgcc ctgcacctgg ctgcccgata ctctcgttca
     1201 gatgctgcca agcgcttgct ggaggccagc gcagatgcca acatccaaga caacatgggg
     1261 cgtaccccat tacatgccgc tgtttctgca gacgctcagg gtgtcttcca gatcctgctc
     1321 cggaacagag ccacagatct ggatgcccga atgcatgatg gcacaacccc tctgatcctg
     1381 gcggcacgcc tggccgtgga aggcatgcta gaggacctca tcaactctca cgctgatgtc
     1441 aatgctgtgg atgacctagg caagtcagct ctgcactggg cagccgctgt gaacaatgtg
     1501 gacgctgctg ttgtgctcct gaagaacgga gccaacaaag acatgcagaa caacaaggag