pMXs-Klf4 vector (V006689)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V006689	pMXs-Klf4	In stock, 1 week for quality controls	Buy one, get one free!

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:: pMXs-Klf4

Antibiotic Resistance:: Ampicillin

Length:: 6210 bp

Type:: Mammalian Expression, Retroviral

Replication origin:: ori

Copy Number:: High Copy

5' Primer:: pMX-S1811 (5'-GACGGCATCGCAGCTTGGATACAC-3')

3' Primer:: LNCX-3 (5'-ACCTACAGGTGGGGTCTTTCATTCCC-3')

pMXs-Klf4 vector Map

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Plasmid 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 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

pMXs-Klf4 vector Sequence

LOCUS       40924_32903        6210 bp DNA     circular SYN 13-MAY-2021
DEFINITION  Retroviral expression of mouse Klf4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6210)
  AUTHORS   Takahashi K, Yamanaka S
  TITLE     Induction of pluripotent stem cells from mouse embryonic and adult 
            fibroblast cultures by defined factors.
  JOURNAL   Cell. 2006 Aug 25. 126(4):663-76.
  PUBMED    16904174
REFERENCE   2  (bases 1 to 6210)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6210)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2006 
            Aug 25. 126(4):663-76."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             complement(120..711)
                     /label=LTR
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     protein_bind    906..930
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(968..2389)
                     /codon_start=1
                     /label=mKlf4
                     /note="Mouse Kruppel-like factor (Klf4) gene. Belongs to
                     the relatively large family of SP1-like transcription 
                     factors and is involved in the regulation of proliferation,
                     differentiation, apoptosis and somatic cell reprogramming."
                     /translation="MAVSDALLPSFSTFASGPAGREKTLRPAGAPTNRWREELSHMKRL
                     PPLPGRPYDLAATVATDLESGGAGAACSSNNPALLARRETEEFNDLLDLDFILSNSLTH
                     QESVAATVTTSASASSSSSPASSGPASAPSTCSFSYPIRAGGDPGVAASNTGGGLLYSR
                     ESAPPPTAPFNLADINDVSPSGGFVAELLRPELDPVYIPPQQPQPPGGGLMGKFVLKAS
                     LTTPGSEYSSPSVISVSKGSPDGSHPVVVAPYSGGPPRMCPKIKQEAVPSCTVSRSLEA
                     HLSAGPQLSNGHRPNTHDFPLGRQLPTRTTPTLSPEELLNSRDCHPGLPLPPGFHPHPG
                     PNYPPFLPDQMQSQVPSLHYQELMPPGSCLPEEPKPKRGRRSWPRKRTATHTCDYAGCG
                     KTYTKSSHLKAHLRTHTGEKPYHCDWDGCGWKFARSDELTRHYRKHTGHRPFQCQKCDR
                     AFSRSDHLALHMKRHF"
     protein_bind    complement(2410..2434)
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    complement(2485..2859)
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     CDS             complement(2869..3285)
                     /codon_start=1
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
                     /translation="GQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPTF
                     NVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKP
                     PPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGA"
     misc_feature    complement(3344..3701)
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus
                     (MMLV)"
     promoter        4312..4416
                     /label=AmpR promoter
     CDS             4417..5274
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5441..6029
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6183..6200
                     /label=L4440
                     /note="L4440 vector, forward primer"