pPKm-230 vector (V004066)

MSDS

Basic Vector Information

Vector Name:: pPKm-230

Antibiotic Resistance:: Ampicillin

Length:: 12253 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-230 vector Vector Map

View Map in NovoBuilder

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-230 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V004066                12253 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004066
VERSION     V004066
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 12253)
  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 12253)
  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 12253)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 12253)
  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..12253
                     /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..2832
                     /gene="PIF3"
                     /label="Transcription factor PIF3"
                     /note="Transcription factor PIF3 from Arabidopsis thaliana.
                     Accession#: O80536"
     misc_feature    2842..3017
                     /label="MTAD"
                     /note="MTAD"
     CDS             3022..3033
                     /label="Factor Xa site"
                     /note="Factor Xa recognition and cleavage site"
     CDS             3049..3069
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             3082..3108
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     misc_feature    3121..3707
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3720..5584
                     /label="PhyB"
                     /note="PhyB"
     CDS             5595..5615
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             5625..6065
                     /label="GAL4 DNA binding domain"
                     /note="DNA binding domain of the GAL4 transcriptional
                     activator"
     promoter        6138..6467
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             6477..7073
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     LTR             7369..7602
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7759..7983
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      8026..8035
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        8079..8183
                     /label="AmpR promoter"
     CDS             8184..9041
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      9215..9803
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     promoter        10002..11179
                     /label="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     misc_feature    11260..11493
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             11678..11722
                     /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             11871..11912
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    12009..12126
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"

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