L071 RRI-Green vector (V009888)

Basic Vector Information

      • Vector Name:
      • L071 RRI-Green
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 8254 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Micklem DR, Lorens JB.

L071 RRI-Green vector Vector Map

L071 RRI-Green8254 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 ℃.

L071 RRI-Green vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                8254 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector L071 RRI-Green, complete sequence.
ACCESSION   EU424172
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8254)
  AUTHORS   Micklem DR, Lorens JB.
  TITLE     Retroviral vectors for expression of shRNAs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8254)
  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 8254)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8254
                     /organism="Cloning vector L071 RRI-Green"
                     /mol_type="other DNA"
                     /db_xref="taxon:517185"
     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..3742,4666..4698))
                     /product="GFP2APuro fusion protein"
                     /label=GFP2APuro fusion protein mRNA
     polyA_site      complement(2157)
     CDS             complement(2290..3708)
                     /codon_start=1
                     /product="GFP2APuro fusion protein"
                     /label=GFP2APuro fusion protein
                     /note="fusion between EGFP, FMDV 2A-related self-cleaving 
                     peptide and puromycin acetyl transferase; selectable marker
                     conferring green fluorescence and resistance to puromycin"
                     /protein_id="ACB70965.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKEFGGGSGGGQLLNFDLLKLAGDVESNPGPTTSMTEYKPTVRL
                     ATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERVTELQELFLTRVGLDIGKVWVADD
                     GAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAAQQQMEGLLAPHRPKEPAWFLATV
                     GVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSAPRNLPFYERLGFTVTADVEVPEG
                     PRDLVHDPQARCLI"
     misc_feature    complement(2901..2985)
                     /label=FMDV 2A self-cleaving linker region
                     /note="FMDV 2A self-cleaving linker region"
     CDS             complement(2992..3708)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      complement(3734..4901)
                     /regulatory_class="promoter"
                     /note="EF1-alpha (elongation factor 1 alpha) promoter and 
                     first intron"
     promoter        complement(3734..4896)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          complement(3743..4665)
                     /note="EF1-alpha intron"
     misc_difference complement(4543..4589)
                     /note="not identical to canonical EF1-alpha intron 
                     sequence"
     regulatory      complement(4722..4728)
                     /regulatory_class="TATA_box"
     repeat_region   4962..5869
                     /note="SIN self-inactivating LTR with U6 expression 
                     cassette"
     regulatory      5089..5354
                     /regulatory_class="promoter"
                     /note="modified human U6 promoter"
     protein_bind    5307..5325
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     misc_binding    5307..5324
                     /label=Tet repressor binding site
                     /bound_moiety="Tet repressor"
                     /note="TetO binding site"
     misc_feature    5355..5745
                     /note="stuffer fragment; lost during cloning of shRNAs 
                     between unique BglII and AarI sites"
     misc_feature    5355
                     /label=U6 promoter-dependent transcription initiation
                     /note="U6 promoter-dependent transcription initiation"
     misc_feature    5802..5869
                     /label=R region
                     /note="R region"
     protein_bind    6055..6071
                     /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(6079..6109)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6124..6145
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6418..7130)
                     /direction=LEFT
                     /label=pMB1 ori (pUC)
                     /note="pMB1 ori (pUC)"
     rep_origin      complement(6433..7021)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7195..8055)
                     /codon_start=1
                     /gene="bla"
                     /product="bla"
                     /label=bla
                     /note="beta-lactamase; ampicillin resistance"
                     /protein_id="ACB70964.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(7195..8055)
                     /gene="bla"
                     /label=bla
     promoter        complement(8056..8160)
                     /gene="bla"
                     /label=AmpR promoter

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