Basic Vector Information
- Vector Name:
- pPKm-145
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7573 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-145 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-145 vector Sequence
LOCUS V004073 7573 bp DNA circular SYN 18-DEC-2018 DEFINITION Exported. ACCESSION V004073 VERSION V004073 KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct . REFERENCE 1 (bases 1 to 7573) 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 7573) 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 7573) TITLE Direct Submission REFERENCE 4 (bases 1 to 7573) 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..7573 /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" misc_feature 1281..1867 /label="IRES2" /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 1887..2483 /label="PuroR" /note="puromycin N-acetyltransferase" LTR 2689..2922 /label="3' LTR (Delta-U3)" /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 3079..3303 /label="bGH poly(A) signal" /note="bovine growth hormone polyadenylation signal" regulatory 3346..3355 /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" promoter 3399..3503 /label="AmpR promoter" CDS 3504..4361 /label="AmpR" /note="beta-lactamase" rep_origin 4535..5123 /label="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" LTR 5282..5915 /label="3' LTR" /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 5962..6087 /label="HIV-1 Psi" /note="packaging signal of human immunodeficiency virus type 1" misc_feature 6580..6813 /label="RRE" /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 6998..7042 /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 7191..7232 /note="Protein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509" /label="Protein Tat" misc_feature 7329..7446 /label="cPPT/CTS" /note="central polypurine tract and central termination sequence of HIV-1" promoter 7573 /label="EF-1-alpha promoter" /note="strong constitutive promoter for human elongation factor EF-1-alpha"
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