Basic Vector Information
- Vector Name:
- pPKm-245
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11063 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-245 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-245 vector Sequence
LOCUS V004055 11063 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V004055
VERSION V004055
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 11063)
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 11063)
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 11063)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 11063)
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..11063
/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"
misc_feature 1369..1398
/label="MYC tag codon optimized"
/note="MYC tag codon optimized"
CDS 1369..1398
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label="Myc"
/translation="EQKLISEEDL"
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 3049..3114
/label="P2A peptide(1)"
/note="P2A peptide(1)"
CDS 3058..3114
/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 3115..3201
/label="transit peptide mito"
/note="transit peptide mito"
CDS 3115..3189
/codon_start=1
/product="mitochondrial presequence of human cytochrome c
oxidase subunit VIII (Rizutto et al., 1989)"
/label="COX8 presequence"
/translation="MSVLTPLLLRGLTGSARRLPVPRAK"
regulatory 3211..3220
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 3220..3261
/label="V5 tag"
/note="epitope tag from simian virus 5"
CDS 3268..3561
/gene="petF1"
/label="Ferredoxin-1"
/note="Ferredoxin-1 from Synechococcus elongatus.
Accession#: P0A3D0"
misc_feature 3562..3627
/label="P2A"
/note="P2A"
CDS 3571..3627
/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 3628..3714
/label="transit peptide mito(3)"
/note="transit peptide mito(3)"
CDS 3628..3702
/codon_start=1
/product="mitochondrial presequence of human cytochrome c
oxidase subunit VIII (Rizutto et al., 1989)"
/label="COX8 presequence"
/translation="MSVLTPLLLRGLTGSARRLPVPRAK"
CDS 3733..3750
/label="6xHis"
/note="6xHis affinity tag"
misc_feature 3757..4914
/label="FNR THEEB"
/note="FNR THEEB"
promoter 4948..5277
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
CDS 5287..5883
/label="PuroR"
/note="puromycin N-acetyltransferase"
LTR 6179..6412
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 6569..6793
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
regulatory 6836..6845
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
promoter 6889..6993
/label="AmpR promoter"
CDS 6994..7851
/label="AmpR"
/note="beta-lactamase"
rep_origin 8025..8613
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
LTR 8772..9405
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 9452..9577
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
promoter 10002..11063
/label="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
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