Basic Vector Information
- Vector Name:
- pPKm-233
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9267 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Kyriakakis P, Catanho M, Hoffner N, Thavarajah W, Jian-Yu V, Chao SS, Hsu A, Pham V, Naghavian L, Dozier LE, Patrick G, Coleman T.
- Promoter:
- EF-1α
pPKm-233 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 ℃.
pPKm-233 vector Sequence
LOCUS V004063 9267 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V004063
VERSION V004063
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 9267)
AUTHORS Kyriakakis P, Catanho M, Hoffner N, Thavarajah W, Jian-Yu V, Chao
SS, Hsu A, Pham V, Naghavian L, Dozier LE, Patrick G, Coleman T.
TITLE Biosynthesis of Orthogonal Molecules Using Ferredoxin and
Ferredoxin-NADP+ Reductase Systems Enables Genetically Encoded PhyB
Optogenetics
JOURNAL ACS Synth Biol (2018) In press
PUBMED 29301067
REFERENCE 2 (bases 1 to 9267)
AUTHORS Kyriakakis P.
TITLE Direct Submission
JOURNAL Submitted (15-NOV-2017) Bioengineering, UCSD, 9500 Gilman drive,
PFBH room 251 MC0412, La Jolla, CA 92093, United States of America
REFERENCE 3 (bases 1 to 9267)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9267)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
Biol (2018) In press"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(15-NOV-2017) Bioengineering, UCSD, 9500 Gilman drive, PFBH room 251
MC0412, La Jolla, CA 92093, United States of America"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9267
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 1..1179
/label="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
CDS 1279..1353
/label="COX8 presequence"
/note="mitochondrial presequence of human cytochrome c
oxidase subunit VIII (Rizutto et al., 1989)"
regulatory 1375..1384
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1384..1425
/label="V5 tag"
/note="epitope tag from simian virus 5"
CDS 1432..1722
/gene="petF"
/label="Ferredoxin-1"
/note="Ferredoxin-1 from Synechococcus sp. (strain ATCC
27264 / PCC 7002 / PR-6). Accession#: P31965"
CDS 1738..1794
/label="P2A"
/note="2A peptide from porcine teschovirus-1 polyprotein"
misc_feature 1795..1881
/label="transit peptide mito(1)"
/note="transit peptide mito(1)"
CDS 1795..1869
/codon_start=1
/product="mitochondrial presequence of human cytochrome c
oxidase subunit VIII (Rizutto et al., 1989)"
/label="COX8 presequence"
/translation="MSVLTPLLLRGLTGSARRLPVPRAK"
CDS 1900..1917
/label="6xHis"
/note="6xHis affinity tag"
CDS 1924..3129
/gene="petH"
/label="Ferredoxin--NADP reductase"
/note="Ferredoxin--NADP reductase from Synechococcus sp.
(strain ATCC 27264 / PCC 7002 / PR-6). Accession#: P31973"
promoter 3152..3481
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
CDS 3491..4087
/label="PuroR"
/note="puromycin N-acetyltransferase"
LTR 4383..4616
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4773..4997
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
regulatory 5040..5049
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
promoter 5093..5197
/label="AmpR promoter"
CDS 5198..6055
/label="AmpR"
/note="beta-lactamase"
rep_origin 6229..6817
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
LTR 6976..7609
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 7656..7781
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 8274..8507
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 8692..8736
/label="gp41 peptide"
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
CDS 8885..8926
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 9023..9140
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 9267
/label="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
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