Basic Vector Information
- Vector Name:
- pPICZalphaH6E
- Antibiotic Resistance:
- Bleomycin
- Length:
- 3551 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Janakiraman VN, Cabanne C, Dieryck W, Hocquellet A, Joucla G, Le Senechal C, Chaignepain S, Costaglioli P, Santarelli X, Garbay B, Noubhani A.
- Promoter:
- AOX1
pPICZalphaH6E 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 ℃.
pPICZalphaH6E vector Sequence
LOCUS 40924_34600 3551 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pPICZalphaH6E, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3551)
AUTHORS Janakiraman VN, Cabanne C, Dieryck W, Hocquellet A, Joucla G, Le
Senechal C, Chaignepain S, Costaglioli P, Santarelli X, Garbay B,
Noubhani A.
TITLE Production and purification of recombinant human hepcidin-25 with
authentic N and C-termini
JOURNAL J. Biotechnol. 195, 89-92 (2015)
PUBMED 25562424
REFERENCE 2 (bases 1 to 3551)
AUTHORS Dieryck W, Narasimhan Janakiraman V, Cabanne C, Hocquellet A, Joucla
G, Le Senechal C, Chaignepain S, Costaglioli P, Santarelli X, Garbay
B, Noubhani A.
TITLE Direct Submission
JOURNAL Submitted (19-JUN-2014) ENSTBB, Institut Polytechnique de Bordeaux,
146 rue Leo Saignat, Bordeaux, gironde 33076, France
REFERENCE 3 (bases 1 to 3551)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3551)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J.
Biotechnol. 195, 89-92 (2015)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(19-JUN-2014) ENSTBB, Institut Polytechnique de Bordeaux, 146 rue
Leo Saignat, Bordeaux, gironde 33076, France"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..3551
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 2..940
/label=AOX1 promoter
/note="inducible promoter, regulated by methanol"
CDS 941..1207
/codon_start=1
/label=alpha-factor secretion signal
/note="N-terminal secretion signal from S. cerevisiae
alpha-factor"
/translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE
GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA"
CDS 1238..1255
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
CDS 1301..1315
/codon_start=1
/label=enterokinase site
/note="enterokinase recognition and cleavage site"
/translation="DDDDK"
misc_feature 1315..1359
/label=multiple cloning site
/note="multiple cloning site"
terminator 1375..1621
/label=AOX1 terminator
/note="transcription terminator for AOX1"
promoter 1636..2047
/label=TEF1 promoter
/note="promoter for EF-1-alpha"
promoter 2055..2102
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 2121..2492
/codon_start=1
/label=BleoR
/note="antibiotic-binding protein"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
terminator 2561..2808
/label=CYC1 terminator
/note="transcription terminator for CYC1"
rep_origin complement(2883..3471)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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