GWluc-basic vector (V009927)

Basic Vector Information

      • Vector Name:
      • GWluc-basic
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 6531 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Del Vecchio I, Zuccotti A, Canneva F, Lenzken SC, Racchi M.

GWluc-basic vector Vector Map

GWluc-basic6531 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300multiple cloning site 1AttR1; recombination site for Gateway systemlac UV5 promoterCmRCcdBAttR2; recombination site for Gateway systemmultiple cloning site 2vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)SV40 polyA signal for luc+ reporter transcription terminationmultiple cloning site 3oribeta-lactamaseAmpR promoterf1 originsynthetic polyA signal for transcriptional pause andbackground reduction

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

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                6531 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector GWluc-basic, complete sequence.
ACCESSION   EF450830
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  JOURNAL   Unpublished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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6531
                     /organism="Cloning vector GWluc-basic"
                     /mol_type="other DNA"
                     /label=derived from pGL3-Basic backbone
                     /note="derived from pGL3-Basic backbone"
                     /db_xref="taxon:433691"
     misc_feature    1..28
                     /label=multiple cloning site 1
                     /note="multiple cloning site 1"
     misc_recomb     33..157
                     /note="AttR1; recombination site for Gateway system"
     protein_bind    33..157
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /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..925
                     /codon_start=1
                     /product="CmR"
                     /label=CmR
                     /note="confers chloramphenicol resistance"
                     /protein_id="ABO76903.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             1267..1572
                     /codon_start=1
                     /product="CcdB"
                     /label=CcdB
                     /note="prokaryotic negative selection protein"
                     /protein_id="ABO76904.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     misc_recomb     1613..1737
                     /note="AttR2; recombination site for Gateway system"
     protein_bind    complement(1613..1737)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    1742..1770
                     /label=multiple cloning site 2
                     /note="multiple cloning site 2"
     regulatory      1795..1804
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1801..3453
                     /codon_start=1
                     /gene="luc+"
                     /product="luciferase"
                     /label=luc+
                     /note="derived from Photinus pyralis"
                     /protein_id="ABO76905.1"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     gene            1801..3453
                     /gene="luc+"
                     /label=luc+
     regulatory      3485..3706
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 polyA signal for luc+ reporter transcription 
                     termination"
     polyA_signal    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(4793..5653)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /note="AmpR; confers ampicillin resistance"
                     /protein_id="ABO76906.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5654..5758)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5785..6240
                     /label=f1 origin
                     /note="f1 origin"
     regulatory      6371..6448
                     /regulatory_class="polyA_signal_sequence"
                     /note="synthetic polyA signal for transcriptional pause and
                     background reduction"
     polyA_signal    6371..6419
                     /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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