Basic Vector Information
- Vector Name:
- MStart-36xMashTag-BFP-3xStop-24xPP7 (Moonstart-MashTag reporter)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11144 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells
- Selection Marker:
- Zeo
- Promoter:
- CMV
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
MStart-36xMashTag-BFP-3xStop-24xPP7 (Moonstart-MashTag reporter) vector Vector Map
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 ℃.
MStart-36xMashTag-BFP-3xStop-24xPP7 (Moonstart-MashTag reporter) vector Sequence
LOCUS 62056_1700 11144 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 11144)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..11144
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
protein_bind 820..838
/label=tet operator
/note="bacterial operator O2 for the tetR and tetA genes"
CDS 1025..1081
/codon_start=1
/label=GCN4_v4
/note="GCN4 peptide optimized to improve solubility while
preserving binding to an scFv-GFP fusion protein (Tanenbaum
et al., 2014)"
/translation="EELLSKNYHLENEVARLKK"
CDS 1083..1127
/codon_start=1
/label=gp41 peptide
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
/translation="KNEQELLELDKWASL"
CDS 4861..5556
/codon_start=1
/label=TagBFP
/note="monomeric blue fluorescent protein"
/translation="SELIRENMHIKLYMEGTVDNHHFKCTSEGEGKPYEGTQTIRIKVV
EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPQGFTWERVTTYEDGGVLTAT
QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
LPSKLGHKLN"
polyA_signal 7158..7382
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 7428..7856
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 7870..8199
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
promoter 8247..8294
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 8313..8684
/codon_start=1
/label=BleoR
/note="antibiotic-binding protein"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 8817..8950
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter complement(9035..9065)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(9080..9101)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(9389..9977)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(10151..11008)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(11009..11113)
/label=AmpR promoter
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