Search Name
pECMV-3*Flag-HMGB1-m,pECMV-3*Flag-HMGB1-m vector, pECMV-3*Flag-HMGB1-m plasmid, pECMV-3*Flag-HMGB1-m map
pECMV-3*Flag-HMGB1-m Informaiton
Promoter: ECMV promoter
Replicator: pUC ori, F1 ori, SV40 ori
Terminator: bGH poly (A) signal
Plasmid classification: gene library plasmid, mouse gene plasmid, mouse eukaryotic plasmid.
Plasmid size: 6100bp
Plasmid label: N-3 x FLAG
Prokaryotic resistance: Amp (100 mu g/ml)
Screening markers: Neo
Cloned strain: DH5 alpha
Culture conditions: 37 centigrade, LB, aerobic
Expression host: mammalian cells such as 293T
Culture conditions: 37 C, 5%CO2
Induction mode: no induction
5'sequencing primers: CMV-F (CGCAAATGGGCGGTAGGCGTG)
3'sequencing primers: BGH-R (TAGAAGGCACAGTCGAGG)
pECMV-3*Flag-HMGB1-m Description
The pECMV-3 * FLAG-HMGB1-m plasmid was transformed on the basis of pECMV-3 x FLAG vector. CMV started the overexpression of HMGB1 gene in the mouse source, and the 3 x FLAG fusion label on the N end could be recognized by the FLAG antibody. The plasmid can be directly transfected into mammalian cells as an overexpression plasmid. The plasmid itself has a Neo selection marker, and G418 can be used to screen stable transfected cell lines after transfection.
Gene information from NCBI
Mus musculus high mobility group box 1 (Hmgb1), transcript variant 1, mRNA锛汵CBI Reference Sequence: NM_001313894.1
pECMV-3*Flag-HMGB1-m Sequence
LOCUS Exported 6100 bp ds-DNA circular SYN 10-涓冩湀-2017
KEYWORDS pECMV-3×FLAG-HMGB1-m
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6100)
AUTHORS liutao
TITLE Direct Submission
JOURNAL Exported 2017-7-10
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6100
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 235..614
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 863..881
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 922..987
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/note="3xFLAG"
/translation="DYKDHDGDYKDHDIDYKDDDDK"
misc_feature 1009..1656
/note="HMGB1-m"
polyA_signal 1700..1924
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
rep_origin 1970..2398
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2412..2741
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 2592..2727
/note="SV40 ori"
/note="SV40 origin of replication"
CDS 2808..3602
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="Neo"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 3776..3897
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(4348..4933)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5104..5964)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5965..6069)
/gene="bla"
/note="AmpR promoter"
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
901 gtttaaactt aagccaccat ggactacaaa gaccatgacg gtgattataa agatcatgac
961 atcgactaca aggatgacga tgacaagctt ggtaccgagc tcggatccat gggcaaagga
1021 gatcctaaaa agccgagagg caaaatgtcc tcatatgcat tctttgtgca aacttgccgg
1081 gaggagcaca agaagaagca cccggatgct tctgtcaact tctcagagtt ctccaagaag
1141 tgctcagaga ggtggaagac catgtctgct aaagaaaagg ggaaatttga agatatggca
1201 aaggctgaca aggctcgtta tgaaagagaa atgaaaacct acatcccccc caaaggggag
