L087 RRI-Red vector (V009887)

Basic Vector Information

      • Vector Name:
      • L087 RRI-Red
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 8215 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Micklem DR, Lorens JB.

L087 RRI-Red vector Vector Map

L087 RRI-Red8215 bp400800120016002000240028003200360040004400480052005600600064006800720076008000CRU5 chimeric CMV promoter, MoMLV and MSCV 5' LTRpsi+; chimeric MoMLV/MSCV extended packaging signalBGHpA signal; bovine growth hormone polyadenylation signalEF1-alpha (elongation factor 1 alpha) promoter and first intronSIN self-inactivating LTR with U6 expression cassettelac operatorlac promoterCAP binding sitepMB1 ori (pUC)blaAmpR promoter

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 ℃.

L087 RRI-Red vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                8215 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector L087 RRI-Red, complete sequence.
ACCESSION   EU424173
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8215)
  AUTHORS   Micklem DR, Lorens JB.
  TITLE     Retroviral vectors for expression of shRNAs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8215)
  AUTHORS   Micklem DR, Lorens JB.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-JAN-2008) Biomedicine, University of Bergen, Jonas 
            Lies vei 91, Bergen 5009, Norway
REFERENCE   3  (bases 1 to 8215)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8215
                     /organism="Cloning vector L087 RRI-Red"
                     /mol_type="other DNA"
                     /db_xref="taxon:517186"
     repeat_region   2..747
                     /label=CRU5 chimeric CMV promoter, MoMLV and MSCV 5' LTR
                     /note="CRU5 chimeric CMV promoter, MoMLV and MSCV 5' LTR"
     regulatory      2..571
                     /regulatory_class="enhancer"
                     /note="CMV immediate-early enhancer region"
     enhancer        63..366
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        367..570
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    602..669
                     /label=R region
                     /note="R region"
     misc_feature    748..1665
                     /note="psi+; chimeric MoMLV/MSCV extended packaging signal"
     CDS             1240..1656
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     misc_feature    complement(2063..2270)
                     /note="BGHpA signal; bovine growth hormone polyadenylation 
                     signal"
     polyA_signal    2063..2270
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     mRNA            complement(join(2157..3704,4627..4659))
                     /product="mRFP12APuro fusion protein"
                     /label=mRFP12APuro fusion protein mRNA
     polyA_site      complement(2157)
     CDS             complement(2290..3669)
                     /codon_start=1
                     /product="mRFP12APuro fusion protein"
                     /label=mRFP12APuro fusion protein
                     /note="fusion between mRFP1, FMDV 2A-related self-cleaving 
                     peptide and puromycin acetyl transferase; selectable marker
                     conferring red fluorescence and resistance to puromycin"
                     /protein_id="ACB70967.1"
                     /translation="MVASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIK
                     MRLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRH
                     STGAEFGGGSGGGQLLNFDLLKLAGDVESNPGPTTSMTEYKPTVRLATRDDVPRAVRTL
                     AAAFADYPATRHTVDPDRHIERVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESV
                     EAGAVFAEIGPRMAELSGSRLAAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGS
                     AVVLPGVEAAERAGVPAFLETSAPRNLPFYERLGFTVTADVEVPEGPRDLVHDPQARCL
                     I"
     misc_feature    complement(2901..2985)
                     /label=FMDV 2A self-cleaving linker region
                     /note="FMDV 2A self-cleaving linker region"
     CDS             complement(2992..3669)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MVASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIK
                     MRLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRH
                     STGA"
     regulatory      3662..3671
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     regulatory      complement(3695..4862)
                     /regulatory_class="promoter"
                     /note="EF1-alpha (elongation factor 1 alpha) promoter and 
                     first intron"
     promoter        complement(3695..4857)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          complement(3704..4626)
                     /note="EF1-alpha intron"
     misc_difference complement(4504..4550)
                     /note="not identical to canonical EF1-alpha intron 
                     sequence"
     regulatory      complement(4683..4689)
                     /regulatory_class="TATA_box"
     repeat_region   4923..5830
                     /note="SIN self-inactivating LTR with U6 expression 
                     cassette"
     regulatory      5050..5315
                     /regulatory_class="promoter"
                     /note="modified human U6 promoter"
     protein_bind    5268..5286
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     misc_binding    5268..5285
                     /label=Tet repressor binding site
                     /bound_moiety="Tet repressor"
                     /note="TetO binding site"
     misc_feature    5316..5706
                     /note="stuffer fragment; lost during cloning of shRNAs 
                     between unique BglII and AarI sites"
     misc_feature    5316
                     /label=U6 promoter-dependent transcription initiation
                     /note="U6 promoter-dependent transcription initiation"
     misc_feature    5763..5830
                     /label=R region
                     /note="R region"
     protein_bind    6016..6032
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by 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)."
     promoter        complement(6040..6070)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6085..6106
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6379..7091)
                     /direction=LEFT
                     /label=pMB1 ori (pUC)
                     /note="pMB1 ori (pUC)"
     rep_origin      complement(6394..6982)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7156..8016)
                     /codon_start=1
                     /gene="bla"
                     /product="bla"
                     /label=bla
                     /note="beta-lactamase; ampicillin resistance"
                     /protein_id="ACB70966.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(7156..8016)
                     /gene="bla"
                     /label=bla
     promoter        complement(8017..8121)
                     /gene="bla"
                     /label=AmpR promoter

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