Bcr/Abl P210-pLEF vector (V012195)

Basic Vector Information

      • Vector Name:
      • Bcr/Abl P210-pLEF
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 15474 bp
      • Type:
      • Mammalian Expression
      • Selection Marker:
      • Neomycin (select with G418)
      • Copy Number:
      • High Copy
      • Promoter:
      • SV40
      • Cloning Method:
      • Restriction Enzyme
      • 5' Primer:
      • T7 or GST
      • 3' Primer:
      • SP6

Bcr/Abl P210-pLEF vector Vector Map

Bcr/Abl P210-pLEF15474 bp70014002100280035004200490056006300700077008400910098001050011200119001260013300140001470015400SV40 promoterbeta-globin intronpCAG-FT7GSTthrombin siteSP6 promoterBGH-revf1 oriSV40 promoterNeoR/KanRSV40 poly(A) signalM13 revlac operatorlac promoterCAP binding siteL4440oriAmpRAmpR promoterpRS-markerf1 oriIn lacZ gene

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

Bcr/Abl P210-pLEF vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported               15474 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Bcr/Abl P210-pLEF
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15474)
  AUTHORS   Kweon SM, Cho YJ, Minoo P, Groffen J, Heisterkamp N
  TITLE     Activity of the Bcr GTPase-activating domain is regulated through 
            direct protein/protein interaction with the Rho guanine nucleotide 
            dissociation inhibitor.
  JOURNAL   J Biol Chem. 2008 Feb 8;283(6):3023-30. Epub 2007 Dec 10.
  PUBMED    18070886
REFERENCE   2  (bases 1 to 15474)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Chem."; date: "2008-02-8"; volume: "283(6)"; pages: "3023-30. Epub 
            2007 Dec 10"
FEATURES             Location/Qualifiers
     source          1..15474
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..211
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        23..219
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      70..205
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          250..822
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     830..849
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     877..896
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        877..895
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             911..1564
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1522..1544
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1571..1588
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     promoter        complement(9734..9752)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(9735..9752)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(9772..9789)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9778..10002
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10048..10476
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        10490..10819
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10670..10805
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             10886..11680
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(10940..10959)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     11550..11569
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    11854..11975
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11891..11910)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11945..11964
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12024..12040)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12024..12040)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12037..12059)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12048..12064
                     /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)."
     promoter        complement(12072..12102)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12117..12138
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12255..12272)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12426..13014)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12506..12525)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(13185..14045)
                     /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     13808..13827
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14046..14150)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14225..14244)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(14718..15173)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     15299..15321
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     15313..15330
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     15314..15330
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"

This page is informational only.