pPKm-243 vector (V004057)

Basic Vector Information

Vector Name:
pPKm-243
Antibiotic Resistance:
Ampicillin
Length:
9018 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-243 vector Map

pPKm-2439018 bp40080012001600200024002800320036004000440048005200560060006400680072007600800084008800EF-1-alpha promoterEF-1-alpha promotermOrangeP2A peptidetransit peptide MTS6xHissuperfolder GFPSV40 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

pPKm-243 vector Sequence

LOCUS       V004057                 9018 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004057
VERSION     V004057
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9018)
  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 9018)
  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 9018)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9018)
  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..9018
                     /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..1971
                     /label="mOrange"
                     /note="orange monomeric derivative of DsRed fluorescent
                     protein (Shanet et al., 2004)"
     misc_feature    1972..2037
                     /label="P2A peptide"
                     /note="P2A peptide"
     CDS             1981..2037
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /label="P2A"
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     misc_feature    2038..2124
                     /label="transit peptide MTS"
                     /note="transit peptide MTS"
     CDS             2038..2112
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label="COX8 presequence"
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             2140..2157
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             2170..2883
                     /label="superfolder GFP"
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Pedelacq et al., 2006)"
     promoter        2903..3232
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             3242..3838
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     LTR             4134..4367
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4524..4748
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      4791..4800
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        4844..4948
                     /label="AmpR promoter"
     CDS             4949..5806
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      5980..6568
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     LTR             6727..7360
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    7407..7532
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    8025..8258
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             8443..8487
                     /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             8636..8677
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    8774..8891
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        9018
                     /label="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"

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