Basic Vector Information
- Vector Name:
- pT32_T32_72bp-mKate2
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7277 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Li Y, Jiang Y, Chen H, Liao W, Li Z, Weiss R, Xie Z.
pT32_T32_72bp-mKate2 vector Map
pT32_T32_72bp-mKate2 vector Sequence
LOCUS 40924_42289 7277 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pT32_T32_72bp-mKate2, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7277)
AUTHORS Li Y, Jiang Y, Chen H, Liao W, Li Z, Weiss R, Xie Z.
TITLE Modular construction of mammalian gene circuits using TALE
transcriptional repressors
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7277)
AUTHORS Li Y, Jiang Y, Chen H, Liao W.
TITLE Direct Submission
JOURNAL Submitted (03-SEP-2014) Bioinformatics Division/Center for Synthetic
REFERENCE 3 (bases 1 to 7277)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7277)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(03-SEP-2014) Bioinformatics Division/Center for Synthetic "
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..7277
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 1..21
/label=attB4
/note="core recombination site for the Gateway(R) BP
reaction"
protein_bind 69..163
/label=5X UAS
/note="five tandem copies of the 'ScaI site' 17-mer
CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS)
that efficiently binds yeast Gal4 (Webster et al., 1988;
Pfeiffer et al., 2010)"
protein_bind 177..190
/label=TALER32 bs binding site
/bound_moiety="TALER32 bs"
misc_feature 197..256
/label=CMVmini
/note="CMVmini"
protein_bind 263..276
/label=TALER32 bs binding site
/bound_moiety="TALER32 bs"
protein_bind 293..313
/label=attB1
/note="core recombination site for the Gateway(R) BP
reaction"
CDS 355..372
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
CDS 373..1068
/codon_start=1
/label=mKate2
/note="monomeric far-red fluorescent protein (Shcherbo et
al., 2009)"
/translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
CDLPSKLGHK"
protein_bind complement(1111..1135)
/label=attB2
/note="recombination site for the Gateway(R) BP reaction"
polyA_signal 1290..1345
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(1706..1722)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 1730..1746
/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(1754..1784)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1799..1820
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
misc_feature 1993..2829
/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(2859..2877)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(2898..2914)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(2922..2938)
/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(2946..2976)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(2991..3012)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3300..3888)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4062..4919)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(4920..5024)
/label=AmpR promoter
rep_origin complement(5050..5505)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 5647..5663
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 5673..5691
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 5706..6509
/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 6588..6613
/label=SA
/note="splice acceptor site"
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