pPKm-163 vector (V004072)

Basic Vector Information

Vector Name:
pPKm-163
Antibiotic Resistance:
Ampicillin
Length:
7482 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:
SP6

pPKm-163 vector Map

pPKm-1637482 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300660069007200CMV enhancerCMV promoterT7 promoterTranscription factor PIF3SV40 NLSGAL4 DNA binding domainHASP6 promoterbGH poly(A) signalf1 oriSV40 promoterNeoR/KanRSV40 poly(A) signalM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoter

pPKm-163 vector Sequence

LOCUS       V004072                 7482 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004072
VERSION     V004072
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7482)
  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 7482)
  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 7482)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7482)
  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..7482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..406
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        407..610
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        655..673
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             708..2279
                     /gene="PIF3"
                     /label="Transcription factor PIF3"
                     /note="Transcription factor PIF3 from Arabidopsis thaliana.
                     Accession#: O80536"
     CDS             2292..2312
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             2322..2762
                     /label="GAL4 DNA binding domain"
                     /note="DNA binding domain of the GAL4 transcriptional
                     activator"
     CDS             2778..2804
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     promoter        complement(2826..2844)
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2870..3094
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3140..3568
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3582..3912
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             3979..4770
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    4947..5080
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5117..5133)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(5141..5157)
                     /label="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5165..5195)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5210..5231)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5519..6107)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(6281..7138)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(7139..7243)
                     /label="AmpR promoter"

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