Basic Vector Information
- Vector Name:
- pPIC9K-His-CsnA(Streptomyces)
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9972 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Yeast
- Selection Marker:
- HIS4,Neo/G418
- Promoter:
- AOX1
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
pPIC9K-His-CsnA(Streptomyces) 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 ℃.
pPIC9K-His-CsnA(Streptomyces) vector Sequence
LOCUS 62056_18470 9972 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 9972) AUTHORS . TITLE Direct Submission FEATURES Location/Qualifiers source 1..9972 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 2..937 /label=AOX1 promoter /note="inducible promoter, regulated by methanol" CDS 949..1215 /codon_start=1 /label=alpha-factor secretion signal /note="N-terminal secretion signal from S. cerevisiae alpha-factor" /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA" CDS 1924..1941 /codon_start=1 /label=6xHis /note="6xHis affinity tag" /translation="HHHHHH" terminator 2017..2263 /label=AOX1 terminator /note="transcription terminator for AOX1" CDS complement(5627..6439) /codon_start=1 /label=KanR /note="aminoglycoside phosphotransferase" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" misc_feature 6818..7574 /label=AOX1 3' fragment /note="region downstream of Pichia pastoris AOX1 gene" misc_feature 7717..7857 /label=bom /note="basis of mobility region from pBR322" rep_origin complement(8043..8631) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(8805..9662) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(9663..9767) /label=AmpR promoter
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