Basic Vector Information
- Vector Name:
- w-repeats
- Antibiotic Resistance:
- Ampicillin
- Length:
- 14656 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Akmammedov A, Geigges M, Paro R.
- Promoter:
- hsp70
w-repeats 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 ℃.
w-repeats vector Sequence
LOCUS 40924_49372 14656 bp DNA circular SYN 18-DEC-2018 DEFINITION Cloning vector w-repeats, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 14656) AUTHORS Akmammedov A, Geigges M, Paro R. TITLE Single vector non-leaky gene expression system for Drosophila melanogaster JOURNAL Sci Rep 7 (1), 6899 (2017) PUBMED 28761084 REFERENCE 2 (bases 1 to 14656) AUTHORS Akmammedov A, Geigges M, Paro R. TITLE Direct Submission JOURNAL Submitted (04-MAY-2017) Biosystem Science and Engineering, ETH Zurich, Mattenstrasse 26, Basel 4058, Switzerland REFERENCE 3 (bases 1 to 14656) TITLE Direct Submission REFERENCE 4 (bases 1 to 14656) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep"; date: "2017"; volume: "7"; issue: "1"; pages: "6899" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (04-MAY-2017) Biosystem Science and Engineering, ETH Zurich, Mattenstrasse 26, Basel 4058, Switzerland" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..14656 /mol_type="other DNA" /organism="synthetic DNA construct" rep_origin complement(3..458) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 600..616 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 623..641 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" gene 666..4802 /label=mini-white /note="This modified version of the white gene lacks part of the first intron." protein_bind 4842..5227 /label=gypsy insulator /note="chromatin insulator from Drosophila" polyA_signal complement(5235..5661) /label=hsp70 poly(A) /note="polyadenlyation signal from the Drosophila melanogaster hsp70Ab gene" CDS complement(5680..8724) /codon_start=1 /label=lacZ /note="beta-galactosidase" /translation="VVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRS LNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNVTYPITVNPP FVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYGQDSRLPSEF DLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQISDFHVATR FNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIIDERGGYADRV TLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIENGLLLLNGKP LLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLWYTLCDRYGL YVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWSLGNESGHGA NHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVPKWSIKKWLS LPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSLIKYDENGNP WSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQTIEVTSEYL FRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQLWLTVRVVQ PNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELGNKRWQFNRQ SGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWKAAGHYQAEA ALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDVEVASDTPHP ARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPYVFPSENGLR CGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDGFHMGIGGDD SWSPSVSAELQLSAGRYHYQLVWCQK" promoter complement(8846..9294) /label=hsp70 promoter /note="Drosophila melanogaster hsp70Bb promoter" misc_feature complement(9307..11747) /label=w-repeats /note="w-repeats" protein_bind 11755..12140 /label=gypsy insulator /note="chromatin insulator from Drosophila" protein_bind 12180..12249 /label=attB /note="attB site for the phi-C31 integrase (Groth et al., 2000)" promoter complement(12468..12486) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(12507..12523) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 12531..12547 /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(12555..12585) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(12600..12621) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(12909..13497) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(13671..14528) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(14529..14633) /label=AmpR promoter
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