RN3P-KDM5B-FLAG vector (V006697)

Basic Vector Information

      • Vector Name:
      • RN3P-KDM5B-FLAG
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 7915 bp
      • Type:
      • Mammalian Expression
      • Copy Number:
      • High Copy
      • Promoter:
      • Lac
      • Cloning Method:
      • Gibson Cloning
      • 5' Primer:
      • M13 Reverse
      • 3' Primer:
      • XBG-R (GACTCCATTCGGGTGTTC)

RN3P-KDM5B-FLAG vector Vector Map

RN3P-KDM5B-FLAG7915 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900720075007800pBR322ori-FL4440CAP binding sitelac promoterlac operatorM13 revT3 promoter5'UTR3xFLAGWELQut site3'UTRT7M13 Forwardf1 oriAmpR promoterAmpRori

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 ℃.

RN3P-KDM5B-FLAG vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                7915 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  FLAG-tagged mouse KDM5B cDNA clone for mammalian cell expression.
ACCESSION   .
VERSION     .
KEYWORDS    RN3P-KDM5B-FLAG
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7915)
  AUTHORS   Zhang B, Zheng H, Huang B, Li W, Xiang Y, Peng X, Ming J, Wu X, 
            Zhang Y, Xu Q, Liu W, Kou X, Zhao Y, He W, Li C, Chen B, Li Y, Wang 
            Q, Ma J, Yin Q, Kee K, Meng A, Gao S, Xu F, Na J, Xie W
  TITLE     Allelic reprogramming of the histone modification H3K4me3 in early 
            mammalian development.
  JOURNAL   Nature. 2016 Sep 14;537(7621):553-557. doi: 10.1038/nature19361.
  PUBMED    27626382
REFERENCE   2  (bases 1 to 7915)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Nature"; date: "2016-09-14- 14"; volume: "537"; issue: "7621"; 
            pages: "553-557"
FEATURES             Location/Qualifiers
     source          1..7915
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     432..451
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     685..702
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    819..840
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        855..885
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    893..909
                     /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     898..920
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     917..933
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     917..933
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        954..972
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     5'UTR           997..1039
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     CDS             1069..1134
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1510..1521
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     3'UTR           5841..5966
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(6056..6075)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6057..6075)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6082..6099)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6082..6098)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6091..6113)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      complement(6240..6695)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6377..6398)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6589..6608
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6722..6826
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6827..7687
                     /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(7045..7064)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7858..7915,1..531)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"

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