Basic Vector Information
- Vector Name:
- PiggyBac-EF1a-MCS-EF1a-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7391 bp
- Type:
- Protein expression, Transposition
- Replication origin:
- ori
- Host:
- Mammalian cells
- Selection Marker:
- Puro
- Promoter:
- EF-1α
- Growth Temperature:
- 37℃
PiggyBac-EF1a-MCS-EF1a-Puro 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 ℃.
PiggyBac-EF1a-MCS-EF1a-Puro vector Sequence
LOCUS 62056_12805 7391 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 7391) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..7391 /mol_type="other DNA" /organism="synthetic DNA construct" promoter complement(1..105) /label=AmpR promoter rep_origin complement(131..586) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 728..744 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 1441..2619 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" promoter 2728..2939 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" LTR 2952..3220 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" CDS 3255..3851 /codon_start=1 /label=PuroR /note="puromycin N-acetyltransferase" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" polyA_signal 3962..4094 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5370..5386) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(5394..5410) /label=lac operator /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)." promoter complement(5418..5448) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(5463..5484) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(5772..6360) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6534..7391) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW"
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