Basic Vector Information
- Vector Name:
- pPKm-244
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11599 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-244 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-244 vector Sequence
LOCUS V004056 11599 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V004056
VERSION V004056
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11599)
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 11599)
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 11599)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 11599)
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..11599
/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..1398
/label="Myc"
/note="Myc (human c-Myc proto-oncogene) epitope tag"
misc_feature 1399..2127
/label="tHO1"
/note="tHO1"
CDS 2137..2193
/label="P2A"
/note="2A peptide from porcine teschovirus-1 polyprotein"
misc_feature 2194..2280
/label="transit peptide mito(2)"
/note="transit peptide mito(2)"
CDS 2194..2268
/codon_start=1
/product="mitochondrial presequence of human cytochrome c
oxidase subunit VIII (Rizutto et al., 1989)"
/label="COX8 presequence"
/translation="MSVLTPLLLRGLTGSARRLPVPRAK"
CDS 2296..2319
/label="FLAG"
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
misc_feature 2326..3048
/label="tPCYA"
/note="tPCYA"
regulatory 2760..2769
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
misc_feature 3058..3644
/label="IRES2"
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 3651..3725
/label="COX8 presequence"
/note="mitochondrial presequence of human cytochrome c
oxidase subunit VIII (Rizutto et al., 1989)"
regulatory 3747..3756
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 3756..3797
/label="V5 tag"
/note="epitope tag from simian virus 5"
CDS 3804..4097
/gene="petF1"
/label="Ferredoxin-1"
/note="Ferredoxin-1 from Synechococcus elongatus.
Accession#: P0A3D0"
CDS 4107..4163
/label="P2A"
/note="2A peptide from porcine teschovirus-1 polyprotein"
misc_feature 4164..4250
/label="transit peptide mito(3)"
/note="transit peptide mito(3)"
CDS 4164..4238
/codon_start=1
/product="mitochondrial presequence of human cytochrome c
oxidase subunit VIII (Rizutto et al., 1989)"
/label="COX8 presequence"
/translation="MSVLTPLLLRGLTGSARRLPVPRAK"
CDS 4269..4286
/label="6xHis"
/note="6xHis affinity tag"
misc_feature 4293..5450
/label="FNR THEEB"
/note="FNR THEEB"
promoter 5484..5813
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
CDS 5823..6419
/label="PuroR"
/note="puromycin N-acetyltransferase"
LTR 6715..6948
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 7105..7329
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
regulatory 7372..7381
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
promoter 7425..7529
/label="AmpR promoter"
CDS 7530..8387
/label="AmpR"
/note="beta-lactamase"
rep_origin 8561..9149
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
LTR 9308..9941
/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 11217..11258
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 11355..11472
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
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