pMaster1 vector (V016806)

Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V016806	pMaster1	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:: pMaster1

Antibiotic Resistance:: Ampicillin

Length:: 21149 bp

Type:: Protein expression, Stem cell reprogramming

Replication origin:: ori

Host:: Mammalian cells

Selection Marker:: Neo/G418

Promoter:: CAG

Growth Strain(s):: Stbl3

Growth Temperature:: 37℃

pMaster1 vector Map

View Map in NovoBuilder

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.

pMaster1 vector Sequence

Copy Sequence

Download GenBank File(.gb)

LOCUS       62056_16505       21149 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 21149)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..21149
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1978..3767
                     /label=oriP
                     /note="Epstein-Barr virus oriP replication origin (Yates et
                     al., 2000)"
     promoter        4243..5421
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             5435..6514
                     /codon_start=1
                     /label=hOct4
                     /note="Homo sapiens Oct-4 gene. Encodes a transcription
                     factor containing a POU homeodomain that plays a key role 
                     in embryonic development and stem cell pluripotency. 
                     Aberrant expression in adult tissues is associated with 
                     tumorigenesis."
                     /translation="MAGHLASDFAFSPPPGGGGDGPGGPEPGWVDPRTWLSFQGPPGGP
                     GIGPGVGPGSEVWGIPPCPPPYEFCGGMAYCGPQVGVGLVPQGGLETSQPEGEAGVGVE
                     SNSDGASPEPCTVTPGAVKLEKEKLEQNPEESQDIKALQKELEQFAKLLKQKRITLGYT
                     QADVGLTLGVLFGKVFSQTTICRFEALQLSFKNMCKLRPLLQKWVEEADNNENLQEICK
                     AETLVQARKRKRTSIENRVRGNLENLFLQCPKPTLQQISHIAQQLGLEKDVVRVWFCNR
                     RQKGKRSSSDYAQREDFEAAGSPFSGGPVSFPLAPGPHFGTPGYGSPHFTALYSSVPFP
                     EGEAFPPVSVTTLGSPMHSN"
     CDS             6590..7999
                     /codon_start=1
                     /label=hKLF4
                     /note="Homo sapiens Kruppel-like factor 4 (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="MAVSDALLPSFSTFASGPAGREKTLRQAGAPNNRWREELSHMKRL
                     PPVLPGRPYDLAAATVATDLESGGAGAACGGSNLAPLPRRETEEFNDLLDLDFILSNSL
                     THPPESVAATVSSSASASSSSSPSSSGPASAPSTCSFTYPIRAGNDPGVAPGGTGGGLL
                     YGRESAPPPTAPFNLADINDVSPSGGFVAELLRPELDPVYIPPQQPQPPGGGLMGKFVL
                     KASLSAPGSEYGSPSVISVSKGSPDGSHPVVVAPYNGGPPRTCPKIKQEAVSSCTHLGA
                     GPPLSNGHRPAAHDFPLGRQLPSRTTPTLGLEEVLSSRDCHPALPLPPGFHPHPGPNYP
                     SFLPDQMQPQVPPLHYQELMPPGSCMPEEPKPKRGRRSWPRKRTATHTCDYAGCGKTYT
                     KSSHLKAHLRTHTGEKPYHCDWDGCGWKFARSDELTRHYRKHTGHRPFQCQKCDRAFSR
                     SDHLALHMKRHF"
     CDS             8063..9013
                     /codon_start=1
                     /label=hSOX2
                     /note="Homo sapiens transcription factor SOX-2 gene.
                     Belongs to the SRY-related HMG-box (SOX) family of 
                     transcription factors, which is involved in the regulation 
                     of embryonic development and in the determination of cell 
                     fate"
                     /translation="MYNMMETELKPPGPQQTSGGGGGNSTAAAAGGNQKNSPDRVKRPM
                     NAFMVWSRGQRRKMAQENPKMHNSEISKRLGAEWKLLSETEKRPFIDEAKRLRALHMKE
                     HPDYKYRPRRKTKTLMKKDKYTLPGGLLAPGGNSMASGVGVGAGLGAGVNQRMDSYAHM
                     NGWSNGSYSMMQDQLGYPQHPGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPT
                     YSMSYSQQGTPGMALGSMGSVVKSEASSSPPVVTSSSHSRAPCQAGDLRDMISMYLPGA
                     EVPEPAAPSRLHMSQHYQSGPVPGTAINGTLPLSHM"
     CDS             9083..10399
                     /codon_start=1
                     /label=c-Myc
                     /note="human c-Myc proto-oncogene"
                     /translation="MPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQQSELQPPAP
                     SEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLEMVTE
                     LLGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPN
                     PARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDS
                     LLSSTESSPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGS
                     PSAGGHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNR
                     KCTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKA
                     TAYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLRNSCA"
     polyA_signal    10526..10647
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(10654..11109)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     enhancer        11235..11538
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11540..11817
                     /label=chicken beta-actin promoter
     CDS             12907..14034
                     /codon_start=1
                     /label=HSV TK
                     /note="herpes simplex virus thymidine kinase"
                     /translation="MASYPGHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRPEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQCGGSWREDWGQLSGTAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRSMHVFILDYDQSPAGCRDAL
                     LQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     misc_feature    14053..14639
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             14640..15440
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    15675..15722
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     intron          16393..17409
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and
                     rabbit beta-globin"
     CDS             17478..18392
                     /codon_start=1
                     /label=hNanog
                     /note="Homo sapiens nanog homeobox gene. Encodes a
                     DNA-binding homeobox-family transcription factor involved 
                     in embryonic stem (ES) cell proliferation, renewal, and 
                     pluripotency."
                     /translation="MSVDPACPQSLPCFEASDCKESSPMPVICGPEENYPSLQMSSAEM
                     PHTETVSPLPSSMDLLIQDSPDSSTSPKGKQPTSAEKSVAKKEDKVPVKKQKTRTVFSS
                     TQLCVLNDRFQRQKYLSLQQMQELSNILNLSYKQVKTWFQNQRMKSKRWQKNNWPKNSN
                     GVTQKASAPTYPSLYSSYHQGCLVNPTGNLPMWSNQTWNNSTWSNQTQNIQSWSNHSWN
                     TQTWCTQSWNNQAWNSPFYNCGEESLQSCMQFQPNSPASDLEAALEAAGEGLNVIQQTT
                     RYFSTPQTMDLFLNYSMNMQPEDV"
     CDS             18402..18467
                     /codon_start=1
                     /label=F2A
                     /note="2A peptide from foot-and-mouth disease virus
                     polyprotein"
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
     CDS             18468..19094
                     /codon_start=1
                     /label=hLIN28A
                     /note="Human lin-28 homolog A gene. Encodes a LIN-28 family
                     RNA-binding protein that acts as a posttranscriptional 
                     regulator of genes involved in development, self-renewal of
                     embryonic stem cells and metabolism. Overexpressed in human
                     embryonic stem cells, primary human tumors and human cancer
                     cell lines."
                     /translation="MGSVSNQQFAGGCAKAAEEAPEEAPEDAARAADEPQLLHGAGICK
                     WFNVRMGFGFLSMTARAGVALDPPVDVFVHQSKLHMEGFRSLKEGEAVEFTFKKSAKGL
                     ESIRVTGPGGVFCIGSERRPKGKSMQKRRSKGDRCYNCGGLDHHAKECKLPPQPKKCHF
                     CQSISHMVASCPLKAQQGPSAQGKPTYFREEEEEIHSPTLLPEAQN"
     rep_origin      complement(19279..19867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(20041..20898)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(20899..21003)
                     /label=AmpR promoter