Basic Vector Information
- Vector Name:
- pPKm-231
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9201 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-231 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-231 vector Sequence
LOCUS V004065 9201 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V004065
VERSION V004065
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 9201)
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 9201)
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 9201)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9201)
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..9201
/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 1264..1338
/label="COX8 presequence"
/note="mitochondrial presequence of human cytochrome c
oxidase subunit VIII (Rizutto et al., 1989)"
regulatory 1360..1369
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1369..1410
/label="V5 tag"
/note="epitope tag from simian virus 5"
CDS 1417..1710
/gene="petF1"
/label="Ferredoxin-1"
/note="Ferredoxin-1 from Synechococcus elongatus.
Accession#: P0A3D0"
CDS 1720..1776
/label="P2A"
/note="2A peptide from porcine teschovirus-1 polyprotein"
misc_feature 1777..1863
/label="transit peptide mito(1)"
/note="transit peptide mito(1)"
CDS 1777..1851
/codon_start=1
/product="mitochondrial presequence of human cytochrome c
oxidase subunit VIII (Rizutto et al., 1989)"
/label="COX8 presequence"
/translation="MSVLTPLLLRGLTGSARRLPVPRAK"
CDS 1882..1899
/label="6xHis"
/note="6xHis affinity tag"
misc_feature 1906..3063
/label="FNR THEEB"
/note="FNR THEEB"
promoter 3086..3415
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
CDS 3425..4021
/label="PuroR"
/note="puromycin N-acetyltransferase"
LTR 4317..4550
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4707..4931
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
regulatory 4974..4983
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
promoter 5027..5131
/label="AmpR promoter"
CDS 5132..5989
/label="AmpR"
/note="beta-lactamase"
rep_origin 6163..6751
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
LTR 6910..7543
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 7590..7715
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 8208..8441
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 8626..8670
/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 8819..8860
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 8957..9074
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 9201
/label="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
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