Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V001172 | aMHC-Puro Rex-Blast | 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:
- aMHC-Puro Rex-Blast
- Antibiotic Resistance:
- Ampicillin
- Length:
- 12703 bp
- Type:
- Lentiviral
- Replication origin:
- ori
- Selection Marker:
- Puromycin ; Blasticidin resistanct marker driven by Rex-1 promoter
- Promoter:
- αMHC(long)
- Cloning Method:
- Restriction Enzyme
aMHC-Puro Rex-Blast vector Map
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.
aMHC-Puro Rex-Blast vector Sequence
LOCUS 40924_250 12703 bp DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12703)
AUTHORS Kita-Matsuo H, Barcova M, Prigozhina N, Salomonis N, Wei K, Jacot
JG, Nelson B, Spiering S, Haverslag R, Kim C, Talantova M, Bajpai R,
Calzolari D, Terskikh A, McCulloch AD, Price JH, Conklin BR, Chen
HS, Mercola M
TITLE Lentiviral vectors and protocols for creation of stable hESC lines
for fluorescent tracking and drug resistance selection of
cardiomyocytes.
JOURNAL PLoS ONE. 2009 . 4(4):e5046.
PUBMED 19352491
REFERENCE 2 (bases 1 to 12703)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 12703)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE.
2009 . 4(4):e5046."
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..12703
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 50..353
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 354..557
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 571..751
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 798..923
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 973..1089
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 1176..6619
/label=alpha-MHC(long)
/note="Mouse alpha-cardiac myosin heavy chain promoter (5.4
kb)"
CDS 6642..7238
/codon_start=1
/label=PuroR
/note="puromycin N-acetyltransferase"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
CDS 7985..8380
/codon_start=1
/label=BSD
/note="blasticidin S deaminase"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
misc_feature 8389..8977
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
misc_feature 8987..9220
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
protein_bind complement(9336..9369)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
LTR 9440..9620
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(9670..9689)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind complement(9903..9922)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 10022..10044
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(10082..10100)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 10168..10272
/label=AmpR promoter
CDS 10273..11130
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 11304..11892
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 12046..12063
/label=L4440
/note="L4440 vector, forward primer"
primer_bind complement(12133..12153)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 12151..12448
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
polyA_signal 12569..12703
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"