Basic Vector Information
- Vector Name:
- pPKm-248
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11558 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-248 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-248 vector Sequence
LOCUS V004054 11558 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V004054
VERSION V004054
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11558)
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 11558)
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 11558)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 11558)
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..11558
/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)"
misc_feature 1351..2061
/label="tPCYA"
/note="tPCYA"
misc_feature 2092..2718
/label="IRES"
/note="IRES"
CDS 2731..2805
/label="COX8 presequence"
/note="mitochondrial presequence of human cytochrome c
oxidase subunit VIII (Rizutto et al., 1989)"
CDS 3553..3609
/label="P2A"
/note="2A peptide from porcine teschovirus-1 polyprotein"
misc_feature 3610..3696
/label="transit peptide mito"
/note="transit peptide mito"
CDS 3610..3684
/codon_start=1
/product="mitochondrial presequence of human cytochrome c
oxidase subunit VIII (Rizutto et al., 1989)"
/label="COX8 presequence"
/translation="MSVLTPLLLRGLTGSARRLPVPRAK"
regulatory 3706..3715
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 3715..3756
/label="V5 tag"
/note="epitope tag from simian virus 5"
CDS 3763..4056
/gene="petF1"
/label="Ferredoxin-1"
/note="Ferredoxin-1 from Synechococcus elongatus.
Accession#: P0A3D0"
misc_feature 4057..4122
/label="P2A"
/note="P2A"
CDS 4066..4122
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label="P2A"
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
misc_feature 4123..4209
/label="transit peptide mito(2)"
/note="transit peptide mito(2)"
CDS 4123..4197
/codon_start=1
/product="mitochondrial presequence of human cytochrome c
oxidase subunit VIII (Rizutto et al., 1989)"
/label="COX8 presequence"
/translation="MSVLTPLLLRGLTGSARRLPVPRAK"
CDS 4228..4245
/label="6xHis"
/note="6xHis affinity tag"
misc_feature 4252..5409
/label="FNR THEEB"
/note="FNR THEEB"
promoter 5443..5772
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
CDS 5782..6378
/label="PuroR"
/note="puromycin N-acetyltransferase"
LTR 6674..6907
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 7064..7288
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
regulatory 7331..7340
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
promoter 7384..7488
/label="AmpR promoter"
CDS 7489..8346
/label="AmpR"
/note="beta-lactamase"
rep_origin 8520..9108
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
LTR 9267..9900
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
promoter 10002..11179
/label="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
CDS 11176..11217
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 11314..11431
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
This page is informational only.