Basic Vector Information
GWluc-basic 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 ℃.
GWluc-basic vector Sequence
LOCUS 40924_1254 6531 bp DNA circular SYN 17-DEC-2018 DEFINITION Cloning vector GWluc-basic, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6531) AUTHORS Del Vecchio I, Zuccotti A, Canneva F, Lenzken SC, Racchi M. TITLE Development of the first Gateway firefly luciferase vector and use of reverse transcriptase in FLOE (Fluorescently Labeled Oligonucleotide Extension) reactions JOURNAL Plasmid 58 (3), 269-274 (2007) PUBMED 17707908 REFERENCE 2 (bases 1 to 6531) AUTHORS Del Vecchio I, Zuccotti A, Canneva F, Lanni C, Lenzken C, Govoni S, Racchi M. TITLE Improved firefly luciferase vector to rapidly recombine promoter regions JOURNAL Unpublished REFERENCE 3 (bases 1 to 6531) AUTHORS Del Vecchio I, Zuccotti A, Canneva F, Lanni C, Lenzken C, Govoni S, Racchi M. TITLE Direct Submission JOURNAL Submitted (22-FEB-2007) Centre of Excellence in Applied Biology - Experimental and Applied Pharmacology Dept., University of Pavia, Via Taramelli 14, Pavia 27100, Italy REFERENCE 4 (bases 1 to 6531) TITLE Direct Submission REFERENCE 5 (bases 1 to 6531) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid"; date: "2007"; volume: "58"; issue: "3"; pages: "269-274" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Unpublished" COMMENT SGRef: number: 3; type: "Journal Article"; journalName: "Submitted (22-FEB-2007) Centre of Excellence in Applied Biology - Experimental and Applied Pharmacology Dept., University of Pavia, Via Taramelli 14, Pavia 27100, Italy" COMMENT SGRef: number: 4; type: "Journal Article" FEATURES Location/Qualifiers source 1..6531 /mol_type="other DNA" /organism="synthetic DNA construct" misc_feature 1..28 /label=multiple cloning site 1 /note="multiple cloning site 1" protein_bind 33..157 /label=attR1 /note="recombination site for the Gateway(R) LR reaction" promoter 182..212 /label=lac UV5 promoter /note="E. coli lac promoter with an 'up' mutation" CDS 266..922 /codon_start=1 /label=CmR /note="chloramphenicol acetyltransferase" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" CDS 1267..1569 /codon_start=1 /label=ccdB /note="CcdB, a bacterial toxin that poisons DNA gyrase" /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" protein_bind complement(1613..1737) /label=attR2 /note="recombination site for the Gateway(R) LR reaction" misc_feature 1742..1770 /label=multiple cloning site 2 /note="multiple cloning site 2" CDS 1801..3450 /codon_start=1 /label=luciferase /note="firefly luciferase" /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV" polyA_signal complement(3494..3615) /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" misc_feature 3717..3728 /label=multiple cloning site 3 /note="multiple cloning site 3" rep_origin 4031 /label=ColE1-derived plasmid replication origin /note="ColE1-derived plasmid replication origin" rep_origin complement(4034..4622) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4796..5653) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(5654..5758) /label=AmpR promoter rep_origin 5785..6240 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" polyA_signal 6371..6419 /label=poly(A) signal /note="synthetic polyadenylation signal" misc_feature 6433..6524 /label=pause site /note="RNA polymerase II transcriptional pause signal from the human alpha-2 globin gene"
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