Basic Vector Information
- Vector Name:
- pEXPR65
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7729 bp
- Type:
- Mammalian expression vector
- Replication origin:
- ori
- Source/Author:
- Siciliano V.
pEXPR65 vector Map
pEXPR65 vector Sequence
LOCUS 40924_18916 7729 bp DNA circular SYN 18-DEC-2018
DEFINITION Mammalian expression vector pEXPR65, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7729)
AUTHORS Siciliano V.
TITLE Engineering modular intracellular protein sensor-actuator devices
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7729)
AUTHORS Siciliano V.
TITLE Direct Submission
JOURNAL Submitted (22-MAR-2018) Biological Engineering, Massachusetts
Institute of Technology, 500 Tech Square, Cambridge, MA 02139, USA
REFERENCE 3 (bases 1 to 7729)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7729)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(22-MAR-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..7729
/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)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
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"
protein_bind 4272..4290
/label=tet operator
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4307..4325
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4343..4361
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4379..4397
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4414..4432
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 4450..4468
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
promoter 4480..4518
/label=minimal CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
protein_bind 4558..4582
/label=attB1
/note="recombination site for the Gateway(R) BP reaction"
CDS 4616..5317
/codon_start=1
/label=TEV protease
/note="tobacco etch virus protease (Kapust et al., 2001)"
/translation="SLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITNKHL
FRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQRE
ERICLVTTNFQTKSMSSMVSDTSCTFPSSDGIFWKHWIQTKDGQCGSPLVSTRDGFIVG
IHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMVKPEEP
FQPVKEATQLMN"
CDS 5732..5749
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
protein_bind complement(5762..5786)
/label=attB2
/note="recombination site for the Gateway(R) BP reaction"
polyA_signal 5941..5996
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(6357..6373)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 6381..6397
/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(6405..6435)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6450..6471
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
misc_feature 6644..7480
/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(7510..7528)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(7549..7565)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(7573..7589)
/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(7597..7627)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(7642..7663)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
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