Basic Vector Information
E1b-GFP-Tol2 vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
E1b-GFP-Tol2 vector Sequence
LOCUS Exported 7602 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS E1b-GFP-Tol2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7602)
AUTHORS Birnbaum RY, Clowney EJ, Agamy O, Kim MJ, Zhao J, Yamanaka T,
Pappalardo Z, Clarke SL, Wenger AM, Nguyen L, Gurrieri F, Everman
DB, Schwartz CE, Birk OS, Bejerano G, Lomvardas S, Ahituv N
TITLE Coding exons function as tissue-specific enhancers of nearby genes.
JOURNAL Genome Res. 2012 Jun;22(6):1059-68. Epub 2012 Mar 22.
PUBMED 22442009
REFERENCE 2 (bases 1 to 7602)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Genome
Res."; date: "2012-06"; volume: "22(6)"; pages: "1059-68. Epub 2012
Mar 22"
FEATURES Location/Qualifiers
source 1..7602
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 49..68
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(287..391)
/gene="bla"
/label=AmpR promoter
rep_origin complement(417..872)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(554..575)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 766..785
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 999..1021
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 1013..1030
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 1014..1030
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 1040..1059
/label=T7
/note="T7 promoter, forward primer"
promoter 1040..1058
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
intron 3555..4127
/label=beta-globin intron
/note="intron from rabbit beta-globin gene"
regulatory 4201..4210
/regulatory_class="other"
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 4207..4926
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 4207..4923
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(4252..4273)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(4513..4532)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 4860..4881
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 4971..5105
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4995..5014)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 5049..5068
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(5775..5791)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5775..5791)
/label=pBluescriptSK
/note="For pBluescript vector"
primer_bind complement(5828..5848)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(5828..5846)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(5867..5883)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5867..5883)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5880..5902)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5891..5907
/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(5915..5945)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5960..5981
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6098..6115)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6269..6857)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6349..6368)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(7028..7602,1..286))
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
This page is informational only.