Ganesh-R1 vector (V009965)

Basic Vector Information

      • Vector Name:
      • Ganesh-R1
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 11450 bp
      • Type:
      • Reporter vector
      • Source/Author:
      • Swanson CI, Hinrichs T, Johnson LA, Zhao Y, Barolo S.

Ganesh-R1 vector Vector Map

Ganesh-R111450 bp500100015002000250030003500400045005000550060006500700075008000850090009500100001050011000P element 3' endwhiteGateway recombination cassette containing ccdB and CmR genes flanked by attR1/attR2 sitesspacer sequence includes partial KanR geneDrosophila melanogaster Hsp70 minimal promoterP element 5' endpUC8 plasmid sequence containing AmpR gene

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 ℃.

Ganesh-R1 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported               11450 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Reporter vector Ganesh-R1, complete sequence.
ACCESSION   EF420136
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11450)
  AUTHORS   Swanson CI, Hinrichs T, Johnson LA, Zhao Y, Barolo S.
  TITLE     A directional recombination cloning system for restriction- and 
            ligation-free construction of GFP, DsRed, and lacZ transgenic 
            Drosophila reporters
  JOURNAL   Gene 408 (1-2), 180-186 (2008)
  PUBMED    18077106
REFERENCE   2  (bases 1 to 11450)
  AUTHORS   Rogers CI, Hinrichs T, Johnson LA, Zhao Y, Barolo S.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2007) Cell & Developmental Biology, University of 
            Michigan Medical School, 3047 BSRB, 109 Zina Pitcher Place, Ann 
            Arboe, MI 48109-2200, USA
REFERENCE   3  (bases 1 to 11450)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11450
                     /organism="Reporter vector Ganesh-R1"
                     /lab_host="Drosophila melanogaster"
                     /mol_type="other DNA"
                     /label=Gateway P element nuclear eGFP reporter vector
                     /note="Gateway P element nuclear eGFP reporter vector"
                     /db_xref="taxon:427136"
     misc_feature    1..233
                     /label=P element 3' end
                     /note="P element 3' end"
     misc_feature    1..159
                     /label=P element 3' end
                     /note="P element 3' end"
     gene            242..4247
                     /gene="white"
                     /allele="+mC"
                     /label=white
     misc_feature    4250..5964
                     /note="Gateway recombination cassette containing ccdB and 
                     CmR genes flanked by attR1/attR2 sites"
     protein_bind    4255..4379
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        4404..4434
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             4488..5147
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             5489..5794
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(5835..5959)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    5965..6712
                     /label=spacer sequence includes partial KanR gene
                     /note="spacer sequence includes partial KanR gene"
     regulatory      6713..6846
                     /regulatory_class="promoter"
                     /note="Drosophila melanogaster Hsp70 minimal promoter"
     gene            6755..7974
                     /gene="DsRed.T4-NLS-SV40t"
                     /label=DsRed.T4-NLS-SV40t
                     /note="derived from pH-Red Stinger"
     mRNA            6755..7932
                     /gene="DsRed.T4-NLS-SV40t"
                     /product="nuclear DsRed.T4"
                     /label=DsRed.T4-NLS-SV40t mRNA
     regulatory      6914..6923
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             6920..7729
                     /codon_start=1
                     /gene="DsRed.T4-NLS-SV40t"
                     /product="nuclear DsRed.T4"
                     /label=DsRed.T4-NLS-SV40t
                     /protein_id="ABN80069.1"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEPSTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFLYKSRHRRHRQRSRSRNRSRSRSSERKRRQRSRSRSSERRRYLYK"
     CDS             6920..7594
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEPSTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     misc_feature    7598..7714
                     /gene="DsRed.T4-NLS-SV40t"
                     /label=Drosophila melanogaster tra gene NLS
                     /note="Drosophila melanogaster tra gene NLS"
     polyA_signal    7852..7973
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    8105..8690
                     /label=P element 5' end
                     /note="P element 5' end"
     misc_feature    8920..10949
                     /label=pUC8 plasmid sequence containing AmpR gene
                     /note="pUC8 plasmid sequence containing AmpR gene"
     rep_origin      complement(8925..9513)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(9684..10544)
                     /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"
     promoter        complement(10545..10649)
                     /gene="bla"
                     /label=AmpR promoter

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