pPKm-241 vector (V004058)

MSDS

Basic Vector Information

Vector Name:: pPKm-241

Antibiotic Resistance:: Ampicillin

Length:: 9075 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-241 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-241 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V004058                 9075 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004058
VERSION     V004058
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9075)
  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 9075)
  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 9075)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9075)
  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..9075
                     /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"
     regulatory      1273..1282
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1282..1323
                     /label="V5 tag"
                     /note="epitope tag from simian virus 5"
     CDS             1330..1620
                     /gene="petF"
                     /label="Ferredoxin-1"
                     /note="Ferredoxin-1 from Synechococcus sp. (strain ATCC
                     27264 / PCC 7002 / PR-6). Accession#: P31965"
     CDS             1636..1692
                     /label="P2A"
                     /note="2A peptide from porcine teschovirus-1 polyprotein"
     CDS             1708..1725
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             1732..2937
                     /gene="petH"
                     /label="Ferredoxin--NADP reductase"
                     /note="Ferredoxin--NADP reductase from Synechococcus sp.
                     (strain ATCC 27264 / PCC 7002 / PR-6). Accession#: P31973"
     promoter        2960..3289
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             3299..3895
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     LTR             4191..4424
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4581..4805
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      4848..4857
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        4901..5005
                     /label="AmpR promoter"
     CDS             5006..5863
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      6037..6625
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     LTR             6784..7417
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    7464..7589
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    8082..8315
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             8500..8544
                     /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             8693..8734
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    8831..8948
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        9075
                     /label="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"

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