Basic Vector Information
- Vector Name:
- pEXPR64
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9917 bp
- Type:
- Mammalian expression vector
- Replication origin:
- ori
- Source/Author:
- Siciliano V.
- Promoter:
- EF-1α
pEXPR64 vector Map
pEXPR64 vector Sequence
LOCUS 40924_18911 9917 bp DNA circular SYN 18-DEC-2018
DEFINITION Mammalian expression vector pEXPR64, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9917)
AUTHORS Siciliano V.
TITLE Engineering modular intracellular protein sensor-actuator devices
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 9917)
AUTHORS Siciliano V.
TITLE Direct Submission
JOURNAL Submitted (02-APR-2018) Biological Engineering, Massachusetts
Institute of Technology, 500 Tech Square, Cambridge, MA 02139, USA
REFERENCE 3 (bases 1 to 9917)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9917)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(02-APR-2018) Biological Engineering, Massachusetts Institute of
Technology, 500 Tech Square, Cambridge, MA 02139, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9917
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(222..810)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(984..1841)
/label=AmpR
/note="beta-lactamase"
promoter complement(1842..1946)
/label=AmpR promoter
rep_origin complement(1972..2427)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 2569..2585
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 2595..2613
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 2628..3431
/label=HA-L
/note="left homology arm from the adeno-associated virus
integration site (AAVS1) within intron 1 of the human
PPP1R12C gene"
misc_feature 3510..3535
/label=SA
/note="splice acceptor site"
protein_bind 4207..4227
/label=attB4
/note="core recombination site for the Gateway(R) BP
reaction"
promoter 4252..5424
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
protein_bind 5446..5470
/label=attB1
/note="recombination site for the Gateway(R) BP reaction"
regulatory 5505..5514
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 5619..5648
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
CDS 6102..6797
/label=mKate2
/note="monomeric far-red fluorescent protein (Shcherbo et
al., 2009)"
CDS 7179..7205
/label=HA
/note="HA (human influenza hemagglutinin) epitope tag"
CDS 7212..7232
/label=TEV site
/note="tobacco etch virus (TEV) protease recognition and
cleavage site"
CDS 7239..7472
/codon_start=1
/gene="UL48"
/product="transcriptional activation domain of herpes
simplex virus protein VP16 (Triezenberg et al., 1988;
Cousens et al., 1989)"
/label=VP16 AD
/translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
CDS 7479..7916
/label=GAL4 DNA binding domain
/note="DNA binding domain of the GAL4 transcriptional
activator"
protein_bind complement(7950..7974)
/label=attB2
/note="recombination site for the Gateway(R) BP reaction"
polyA_signal 8129..8184
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(8545..8561)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 8569..8585
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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(8593..8623)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8638..8659
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
misc_feature 8832..9668
/label=HA-R
/note="right homology arm from the adeno-associated virus
integration site (AAVS1) within intron 1 of the human
PPP1R12C gene"
promoter complement(9698..9716)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(9737..9753)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(9761..9777)
/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(9785..9815)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(9830..9851)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
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