pPKm-240 vector (V004059)

Basic Vector Information

Vector Name:
pPKm-240
Antibiotic Resistance:
Ampicillin
Length:
9015 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-240 vector Vector Map

pPKm-2409015 bp40080012001600200024002800320036004000440048005200560060006400680072007600800084008800EF-1-alpha promotervertebrate consensus sequence for strong initiation of translation (Kozak, 1987)MycHO1 PCC7002 optimizedP2AFLAGPcyASV40 promoterPuroR3' LTR (Delta-U3)bGH poly(A) signalvertebrate consensus sequence for strong initiation of translation (Kozak, 1987)AmpR promoterAmpRori3' LTRHIV-1 PsiRREgp41 peptideProtein TatcPPT/CTS

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-240 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V004059                 9015 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004059
VERSION     V004059
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9015)
  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 9015)
  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 9015)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9015)
  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..9015
                     /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..1311
                     /label="Myc"
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     misc_feature    1312..2031
                     /label="HO1 PCC7002 optimized"
                     /note="HO1 PCC7002 optimized"
     CDS             2041..2097
                     /label="P2A"
                     /note="2A peptide from porcine teschovirus-1 polyprotein"
     CDS             2101..2124
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     misc_feature    2131..2865
                     /label="PcyA"
                     /note="PcyA"
     promoter        2900..3229
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             3239..3835
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     LTR             4131..4364
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4521..4745
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      4788..4797
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        4841..4945
                     /label="AmpR promoter"
     CDS             4946..5803
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      5977..6565
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     LTR             6724..7357
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    7404..7529
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    8022..8255
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             8440..8484
                     /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             8633..8674
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    8771..8888
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        9015
                     /label="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"

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