L188-1 vector (V009885)

Basic Vector Information

      • Vector Name:
      • L188-1
      • Antibiotic Resistance:
      • Kanamycin
      • Length:
      • 6235 bp
      • Type:
      • Expression vector
      • Source/Author:
      • Sommer F, Awazu S, Anton-Erxleben F, Jiang D, Klimovich AV, Klimovich BV, Samoilovich MP, Satou Y, Kruss M, Gelhaus C, Kurn U, Bosch TC, Khalturin K.

L188-1 vector Vector Map

L188-16235 bp30060090012001500180021002400270030003300360039004200450048005100540057006000f1 oriKanRoribomropCAP binding sitelacIlacI promoterT7 promoterlac operatorRBS6xHisthrombin site6xHisT7 terminator

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 ℃.

L188-1 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_1534        6235 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector L188-1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6235)
  AUTHORS   Sommer F, Awazu S, Anton-Erxleben F, Jiang D, Klimovich AV, 
            Klimovich BV, Samoilovich MP, Satou Y, Kruss M, Gelhaus C, Kurn U, 
            Bosch TC, Khalturin K.
  TITLE     Blood System Formation in the Urochordate Ciona intestinalis 
            Requires the Variable Receptor vCRL1
  JOURNAL   Mol. Biol. Evol. 29 (10), 3081-3093 (2012)
  PUBMED    22513285
REFERENCE   2  (bases 1 to 6235)
  AUTHORS   Sommer F, Awazu S, Anton-Erxleben F, Jiang D, Klimovich AV, 
            Klimovich VB, Samoilovich MP, Satou Y, Kruess M, Gelhaus C, Kuern U,
            Bosch TCG., Khalturin K.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-FEB-2011) Zoological Institute, Christian-Albrechts 
            University Kiel, Am Botanischen Garten 9, Kiel, Schleswig-Holstein 
            24118, Germany
REFERENCE   3  (bases 1 to 6235)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6235)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol. Biol. 
            Evol."; date: "2012"; volume: "29"; issue: "10"; pages: "3081-3093"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (16-FEB-2011) Zoological Institute, Christian-Albrechts University 
            Kiel, Am Botanischen Garten 9, Kiel, Schleswig-Holstein 24118, 
            Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(563..1375)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      1497..2085
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2271..2413)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2518..2706)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(3481..3502)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3518..4597)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4598..4675)
                     /label=lacI promoter
     promoter        4984..5002
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5003..5027
                     /label=lac operator
                     /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)."
     RBS             5042..5064
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5083..5100
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             5110..5127
                     /codon_start=1
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
                     /translation="LVPRGS"
     CDS             6079..6096
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      6163..6210
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"

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