Basic Vector Information
- Vector Name:
- pPKm-243
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9018 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-243 vector Map
pPKm-243 vector Sequence
LOCUS V004057 9018 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V004057
VERSION V004057
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 9018)
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 9018)
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 9018)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9018)
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..9018
/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..1971
/label="mOrange"
/note="orange monomeric derivative of DsRed fluorescent
protein (Shanet et al., 2004)"
misc_feature 1972..2037
/label="P2A peptide"
/note="P2A peptide"
CDS 1981..2037
/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 2038..2124
/label="transit peptide MTS"
/note="transit peptide MTS"
CDS 2038..2112
/codon_start=1
/product="mitochondrial presequence of human cytochrome c
oxidase subunit VIII (Rizutto et al., 1989)"
/label="COX8 presequence"
/translation="MSVLTPLLLRGLTGSARRLPVPRAK"
CDS 2140..2157
/label="6xHis"
/note="6xHis affinity tag"
CDS 2170..2883
/label="superfolder GFP"
/note="GFP variant that folds robustly even when fused to
poorly folded proteins (Pedelacq et al., 2006)"
promoter 2903..3232
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
CDS 3242..3838
/label="PuroR"
/note="puromycin N-acetyltransferase"
LTR 4134..4367
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4524..4748
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
regulatory 4791..4800
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
promoter 4844..4948
/label="AmpR promoter"
CDS 4949..5806
/label="AmpR"
/note="beta-lactamase"
rep_origin 5980..6568
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
LTR 6727..7360
/label="3' LTR"
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 7407..7532
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 8025..8258
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 8443..8487
/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 8636..8677
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 8774..8891
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 9018
/label="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
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