Basic Vector Information
- Vector Name:
- pEGFP.attB
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9154 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Bischof J, Bjorklund M, Furger E, Schertel C, Taipale J, Basler K.
- Promoter:
- hsp70
pEGFP.attB 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 ℃.
pEGFP.attB vector Sequence
LOCUS 40924_17249 9154 bp DNA circular SYN 17-DEC-2018 DEFINITION Cloning vector pEGFP.attB, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9154) AUTHORS Bischof J, Bjorklund M, Furger E, Schertel C, Taipale J, Basler K. TITLE A versatile platform for creating a comprehensive UAS-ORFeome library in Drosophila JOURNAL Development 140 (11), 2434-2442 (2013) PUBMED 23637332 REFERENCE 2 (bases 1 to 9154) AUTHORS Bischof J, Bjorklund M, Furger E, Schertel C, Taipale J, Basler K. TITLE Direct Submission JOURNAL Submitted (13-APR-2013) Institute of Molecular Life Sciences, University of Zurich, Winterthurerstrasse 190, Zurich 8057, Switzerland REFERENCE 3 (bases 1 to 9154) TITLE Direct Submission REFERENCE 4 (bases 1 to 9154) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Development"; date: "2013"; volume: "140"; issue: "11"; pages: "2434-2442" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (13-APR-2013) Institute of Molecular Life Sciences, University of Zurich, Winterthurerstrasse 190, Zurich 8057, Switzerland" COMMENT SGRef: number: 3; type: "Journal Article" COMMENT ##Assembly-Data-START## Sequencing Technology :: Sanger dideoxy sequencing ##Assembly-Data-END## FEATURES Location/Qualifiers source 1..9154 /mol_type="other DNA" /organism="synthetic DNA construct" misc_feature 1..4136 /label=white dominant eye marker /note="white dominant eye marker" CDS join(479..550,952..1225,1300..1954,2016..2331,2552..2683, 2754..3368) /codon_start=1 /gene="white" /product="Drosophila white gene eye color pigment" /label=mini-white /note="This ia a modified version of the white gene lacking part of the first intron." /translation="MGQEDQELLIRGGSKHPSAEHLNNGDSGAASQSCINQGFGQAKNY GTLRPPSPPEDSGSGSGQLAENLTYAWHNMDIFGAVNQPGSGWRQLVNRTRGLFCNERH IPAPRKHLLKNVCGVAYPGELLAVMGSSGAGKTTLLNALAFRSPQGIQVSPSGMRLLNG QPVDAKEMQARCAYVQQDDLFIGSLTAREHLIFQAMVRMPRHLTYRQRVARVDQVIQEL SLSKCQHTIIGVPGRVKGLSGGERKRLAFASEALTDPPLLICDEPTSGLDSFTAHSVVQ VLKKLSQKGKTVILTIHQPSSELFELFDKILLMAEGRVAFLGTPSEAVDFFSYVGAQCP TNYNPADFYVQVLAVVPGREIESRDRIAKICDNFAISKVARDMEQLLATKNLEKPLEQP ENGYTYKATWFMQFRAVLWRSWLSVLKEPLLVKVRLIQTTMVAILIGLIFLGQQLTQVG VMNINGAIFLFLTNMTFQNVFATINVFTSELPVFMREARSRLYRCDTYFLGKTIAELPL FLTVPLVFTAIAYPMIGLRAGVLHFFNCLALVTLVANVSTSFGYLISCASSSTSMALSV GPPVIIPFLLFGGFFLNSGSVPVYLKWLSYLSWFRYANEGLLINQWADVEPGEISCTSS NTTCPSSGKVILETLNFSAADLPLDYVGLAILIVSFRVLAYLALRLRARRKE" protein_bind 4143..4176 /label=loxP /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." misc_feature 4189..4258 /label=multiple cloning site /note="multiple cloning site" promoter 4263..4501 /label=hsp70 promoter /note="Drosophila melanogaster hsp70Bb promoter" CDS 4553..5269 /codon_start=1 /label=EGFP /note="enhanced GFP" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" intron 5362..5427 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" CDS 5557..5577 /codon_start=1 /label=SV40 NLS /note="nuclear localization signal of SV40 (simian virus 40) large T antigen" /translation="PKKKRKV" polyA_signal 5849..5983 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" protein_bind 6012..6081 /label=attB /note="attB site for the phi-C31 integrase (Groth et al., 2000)" promoter complement(6300..6318) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(6339..6355) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(6363..6379) /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(6387..6417) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(6432..6453) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(6741..7329) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(7503..8360) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(8361..8465) /label=AmpR promoter rep_origin complement(8491..8946) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 9088..9104 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 9111..9129 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" gene 9154 /label=mini-white /note="This modified version of the white gene lacks part of the first intron."
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