Basic Vector Information
- Vector Name:
- phEF1a-EYFP-2A-Hyg
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9487 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Li Y, Jiang Y, Chen H, Liao W, Li Z, Weiss R, Xie Z.
- Promoter:
- EF-1α
phEF1a-EYFP-2A-Hyg vector Vector Map
phEF1a-EYFP-2A-Hyg vector Sequence
LOCUS 40924_24428 9487 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector phEF1a-EYFP-2A-Hyg, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9487)
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JOURNAL Unpublished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 9487)
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 9487)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9487)
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"Unpublished"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..9487
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 1..21
/label=attB4
/note="core recombination site for the Gateway(R) BP
reaction"
promoter 46..1218
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
protein_bind 1241..1265
/label=attB1
/note="recombination site for the Gateway(R) BP reaction"
CDS 1286..2002
/codon_start=1
/label=mEYFP
/note="enhanced YFP with monomerizing A206K mutation
(Zacharias et al., 2002)"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
misc_feature 2009..2062
/label=2A
/note="2A"
CDS 2009..2062
/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 2069..3103
/codon_start=1
/label=HygR
/note="hygromycin B phosphotransferase"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
MGEAN"
protein_bind complement(3189..3213)
/label=attB2
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(3214..3338)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
polyA_signal 3493..3548
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(3909..3925)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 3933..3949
/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(3957..3987)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4002..4023
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
misc_feature 4196..5032
/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(5062..5080)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(5101..5117)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(5125..5141)
/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(5149..5179)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(5194..5215)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5503..6091)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6265..7122)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(7123..7227)
/label=AmpR promoter
rep_origin complement(7253..7708)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 7850..7866
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 7876..7894
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 7909..8712
/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 8791..8816
/label=SA
/note="splice acceptor site"
This page is informational only.