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E1b-GFP-Tol2 vector (V006993#)

Basic Vector Information

Basic Vector Information
Vector Name E1b-GFP-Tol2 Antibiotic Resistance Ampicillin
Length 7602 bp Type Zebrafish enhancer assay vector
Copy Number High Copy Promoter E1b
Cloning Method Restriction Enzyme 5' Primer GATGCGGGAAGAGGTGTATTAGT
3' Primer AGTTCTCAGGATCGGTCGAAAT

E1b-GFP-Tol2 vector Vector Map

E1b-GFP-Tol27602 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600690072007500Amp-RAmpR promoterf1 oriIn lacZ geneT7beta-globin intronKozak sequenceSV40 poly(A) signalSK primerT3M13 revlac operatorlac promoterCAP binding siteL4440oriAmpR

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

Copy Sequence

Download GeneBank File(.gb)

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"

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