RHP-AmpS vector (V001605)

Basic Vector Information

      • Vector Name:
      • RHP-AmpS
      • Antibiotic Resistance:
      • Kanamycin
      • Length:
      • 6384 bp
      • Type:
      • Expression vector
      • Source/Author:
      • Haag AF, Ostermeier C.

RHP-AmpS vector Vector Map

RHP-AmpS6384 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300f1 oriKan(R)oribomroplacIlacI promoterT7 promoterlac operatorRBSHis6-tagPreScission protease siteBlaD290AmpR promoterT7

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 ℃.

RHP-AmpS vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                6384 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector RHP-AmpS, complete sequence.
ACCESSION   FJ545754
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6384)
  AUTHORS   Haag AF, Ostermeier C.
  TITLE     Positive Selection Vector for Direct Protein Expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6384)
  AUTHORS   Haag AF, Ostermeier C.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-DEC-2008) Novartis Institutes for BioMedical Research,
            Novartis Pharma AG, Forum 1, Novartis Campus, Basel CH-4056, 
            Switzerland
REFERENCE   3  (bases 1 to 6384)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6384
                     /organism="Expression vector RHP-AmpS"
                     /mol_type="other DNA"
                     /db_xref="taxon:660654"
     rep_origin      1..456
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(549..1364)
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /note="Kan(R)"
                     /protein_id="ACT98418.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      1486..2074
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2260..2402
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2504..2695)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3504..4586)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     CDS             complement(3504..4463)
                     /codon_start=1
                     /product="LacI"
                     /label=LacI
                     /protein_id="ACT98419.1"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSR
                     ADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPA
                     LFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKY
                     LTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLR
                     VGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVS
                     LVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ"
     promoter        complement(4587..4664)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        4973..4991
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4973..4989
                     /regulatory_class="promoter"
                     /note="T7"
     protein_bind    4992..5016
                     /label=lac repressor protein binding site
                     /bound_moiety="lac repressor protein"
                     /note="lac operator"
     RBS             5031..5053
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     regulatory      5048..5053
                     /regulatory_class="ribosome_binding_site"
     misc_feature    5228..5245
                     /label=His6-tag
                     /note="His6-tag"
     CDS             5228..5245
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    5249..5272
                     /label=PreScission protease site
                     /note="PreScission protease site"
     CDS             5249..5272
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(5298..6155)
                     /codon_start=1
                     /product="BlaD290"
                     /label=BlaD290
                     /protein_id="ACT98420.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKH"
     promoter        complement(6156..6227)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(6156..6190)
                     /regulatory_class="promoter"
                     /note="bla"
     regulatory      6290..6308
                     /regulatory_class="terminator"
                     /note="T7"
     terminator      6301..6348
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"

This page is informational only.