Basic Vector Information
- Vector Name:
- pTRE-EYFP-2A-TALER9-4xTarget_FF5
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11185 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Li Y, Jiang Y, Chen H, Liao W, Li Z, Weiss R, Xie Z.
- Promoter:
- tight TRE
pTRE-EYFP-2A-TALER9-4xTarget_FF5 vector Map
pTRE-EYFP-2A-TALER9-4xTarget_FF5 vector Sequence
LOCUS 40924_44013 11185 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pTRE-EYFP-2A-TALER9-4xTarget_FF5, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11185)
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JOURNAL Unpublished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 11185)
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 11185)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 11185)
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"Unpublished"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..11185
/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"
promoter 4242..4556
/label=tight TRE promoter
/note="Tet-responsive promoter PTight, consisting of seven
tet operator sequences followed by the minimal CMV
promoter"
protein_bind 4629..4653
/label=attB1
/note="recombination site for the Gateway(R) BP reaction"
CDS 4676..5392
/codon_start=1
/label=mEYFP
/note="enhanced YFP with monomerizing A206K mutation
(Zacharias et al., 2002)"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
misc_feature 5399..5452
/label=2A
/note="2A"
CDS 5399..5452
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
misc_feature 5468..9007
/label=similar to TALER9
/note="similar to TALER9"
CDS 9017..9037
/codon_start=1
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/translation="PKKKRKV"
misc_feature 9097..9118
/label=FF5
/note="FF5"
misc_feature 9119..9140
/label=FF5
/note="FF5"
misc_feature 9141..9162
/label=FF5
/note="FF5"
misc_feature 9163..9184
/label=FF5
/note="FF5"
protein_bind complement(9218..9242)
/label=attB2
/note="recombination site for the Gateway(R) BP reaction"
polyA_signal 9397..9452
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(9813..9829)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 9837..9853
/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(9861..9891)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9906..9927
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
misc_feature 10100..10936
/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(10966..10984)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(11005..11021)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 11029..11045
/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(11053..11083)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(11098..11119)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
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