Basic Vector Information
- Vector Name:
- pCAG-Gal4VP16-2A-TagBFP-2A-Bla
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9890 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Li Y, Jiang Y, Chen H, Liao W, Li Z, Weiss R, Xie Z.
- Promoter:
- chicken β-actin
pCAG-Gal4VP16-2A-TagBFP-2A-Bla vector Vector Map
pCAG-Gal4VP16-2A-TagBFP-2A-Bla vector Sequence
LOCUS 40924_8036 9890 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pCAG-Gal4VP16-2A-TagBFP-2A-Bla, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9890)
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JOURNAL Unpublished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 9890)
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 9890)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9890)
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"Unpublished"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..9890
/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"
enhancer 4313..4616
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 4619..4894
/label=chicken beta-actin promoter
intron 4900..5908
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
protein_bind 5966..5990
/label=attB1
/note="recombination site for the Gateway(R) BP reaction"
regulatory 6010..6019
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 6019..6252
/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 6259..6696
/label=GAL4 DNA binding domain
/note="DNA binding domain of the GAL4 transcriptional
activator"
misc_feature 6697..6762
/label=P2A
/note="P2A"
CDS 6706..6762
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 6769..7464
/label=TagBFP
/note="monomeric blue fluorescent protein"
misc_feature 7465..7518
/label=2A
/note="2A"
CDS 7465..7518
/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"
CDS 7519..7914
/label=BSD
/note="blasticidin S deaminase"
protein_bind complement(7923..7947)
/label=attB2
/note="recombination site for the Gateway(R) BP reaction"
polyA_signal 8102..8157
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(8518..8534)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 8542..8558
/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(8566..8596)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8611..8632
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
misc_feature 8805..9641
/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(9671..9689)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(9710..9726)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(9734..9750)
/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(9758..9788)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(9803..9824)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
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