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 Map
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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