pPKm-244 vector (V004056)

Basic Vector Information

Vector Name:
pPKm-244
Antibiotic Resistance:
Ampicillin
Length:
11599 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-244 vector Map

pPKm-24411599 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000105001100011500EF-1-alpha promoterCOX8 presequenceMyctHO1P2Atransit peptide mito(2)FLAGtPCYAIRES2COX8 presequenceV5 tagFerredoxin-1P2Atransit peptide mito(3)6xHisFNR THEEBSV40 promoterPuroR3' LTR (Delta-U3)bGH poly(A) signalvertebrate consensus sequence for strong initiation of translation (Kozak, 1987)AmpR promoterAmpRori3' LTREF-1-alpha promoterProtein TatcPPT/CTS

pPKm-244 vector Sequence

LOCUS       V004056                11599 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004056
VERSION     V004056
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 11599)
  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 11599)
  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 11599)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 11599)
  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..11599
                     /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..1338
                     /label="COX8 presequence"
                     /note="mitochondrial presequence of human cytochrome c
                     oxidase subunit VIII (Rizutto et al., 1989)"
     regulatory      1360..1369
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1369..1398
                     /label="Myc"
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     misc_feature    1399..2127
                     /label="tHO1"
                     /note="tHO1"
     CDS             2137..2193
                     /label="P2A"
                     /note="2A peptide from porcine teschovirus-1 polyprotein"
     misc_feature    2194..2280
                     /label="transit peptide mito(2)"
                     /note="transit peptide mito(2)"
     CDS             2194..2268
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label="COX8 presequence"
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             2296..2319
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     misc_feature    2326..3048
                     /label="tPCYA"
                     /note="tPCYA"
     regulatory      2760..2769
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3058..3644
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             3651..3725
                     /label="COX8 presequence"
                     /note="mitochondrial presequence of human cytochrome c
                     oxidase subunit VIII (Rizutto et al., 1989)"
     regulatory      3747..3756
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3756..3797
                     /label="V5 tag"
                     /note="epitope tag from simian virus 5"
     CDS             3804..4097
                     /gene="petF1"
                     /label="Ferredoxin-1"
                     /note="Ferredoxin-1 from Synechococcus elongatus.
                     Accession#: P0A3D0"
     CDS             4107..4163
                     /label="P2A"
                     /note="2A peptide from porcine teschovirus-1 polyprotein"
     misc_feature    4164..4250
                     /label="transit peptide mito(3)"
                     /note="transit peptide mito(3)"
     CDS             4164..4238
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label="COX8 presequence"
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             4269..4286
                     /label="6xHis"
                     /note="6xHis affinity tag"
     misc_feature    4293..5450
                     /label="FNR THEEB"
                     /note="FNR THEEB"
     promoter        5484..5813
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             5823..6419
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     LTR             6715..6948
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7105..7329
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      7372..7381
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        7425..7529
                     /label="AmpR promoter"
     CDS             7530..8387
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      8561..9149
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     LTR             9308..9941
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     promoter        10002..11179
                     /label="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             11217..11258
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    11355..11472
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"

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