pLV6-Bmal-luc vector (V012180)

Price Information

Cat No. Plasmid Name Availability Add to cart
V012180 pLV6-Bmal-luc In stock, 1 week for quality controls

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Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:
pLV6-Bmal-luc
Antibiotic Resistance:
Ampicillin
Length:
10234 bp
Type:
Mammalian Expression, Lentiviral, Luciferase
Replication origin:
ori
Selection Marker:
Blasticidin
Copy Number:
High Copy
Promoter:
RSV
Cloning Method:
Restriction Enzyme
3' Primer:
lucNRev

pLV6-Bmal-luc vector Map

pLV6-Bmal-luc10234 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000RSV promoter5' LTR (truncated)HIV-1 PsiRREgp41 peptideProtein TatluciferaseV5 tagSV40 promoterEM7 promoterBSD3' LTR (Delta-U3)SV40 poly(A) signalSV40 oriT7 promoterIn lacZ genef1 oriAmpR promoterAmpRoriL4440CAP binding sitelac promoterlac operatorM13 revT3 promoter

Plasmid 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 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

pLV6-Bmal-luc vector Sequence

LOCUS       V012180                10234 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V012180
VERSION     V012180
KEYWORDS    pLV6-Bmal-luc
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10234)
  TITLE     Steven Brown plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10234)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 10234)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..229
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             230..410
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    457..582
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1075..1308
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1493..1537
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             1686..1727
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     CDS             2954..4603
                     /label="luciferase"
                     /note="firefly luciferase"
     CDS             5744..5785
                     /label="V5 tag"
                     /note="epitope tag from simian virus 5"
     promoter        5827..6156
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     promoter        6204..6251
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     CDS             6270..6665
                     /label="BSD"
                     /note="blasticidin S deaminase"
     LTR             6755..6988
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7060..7194
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      7221..7356
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(7377..7395)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7405..7422)
                     /label="M13 Forward"
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21)
                     Forward"
     primer_bind     complement(7405..7421)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     primer_bind     complement(7414..7436)
                     /label="M13/pUC Forward"
                     /note="In lacZ gene"
     rep_origin      7563..8018
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        8044..8148
                     /label="AmpR promoter"
     CDS             8149..9006
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      9180..9768
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     primer_bind     9922..9939
                     /label="L4440"
                     /note="L4440 vector, forward primer"
     protein_bind    10056..10077
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        10092..10122
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    10130..10146
                     /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)."
     primer_bind     10154..10170
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        10191..10209
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"