YCplac22 5-1.2-FLuciferase vector (V001502) Gene synthesis in YCplac22 5-1.2-FLuciferase backbone

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Price Information

Cat No.	Plasmid Name	Availability	Buy one, get one free! (?)
V001502	YCplac22 5-1.2-FLuciferase	In stock, instant shipping

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

Basic Vector Information

YCplac22 5-1.2-FLuciferase is the YCplac22 with a fLuciferase marker. With the combination of TRP1 promoter and CEN/ARS element, it provides a stable platform for inserting and expressing various target genes. Scientists can insert genes of interest into YCplac22 5-1.2-FLuciferase and introduce it into host organisms, such as bacteria or yeast, to study the function and regulation of those genes. The luciferase provides another way to select the cell that carries the interested gene.

YCplac22 5-1.2-FLuciferase is also useful in constructing mutant libraries. By randomly mutagenizing genes and inserting them into YCplac22 5-1.2-FLuciferase, researchers can screen for mutants with desired phenotypes, helping to understand gene function and identify potential genetic factors involved in certain traits.

Vector Name:: YCplac22 5-1.2-FLuciferase

Antibiotic Resistance:: Ampicillin

Length:: 7428 bp

Type:: Cloning vector

Replication origin:: ori

Source/Author:: Belmont BJ, Niles JC.

Promoter:: TRP1

Growth Strain(s):: DH10B

Growth Temperature:: 37℃

YCplac22 5-1.2-FLuciferase vector Map

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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.

YCplac22 5-1.2-FLuciferase vector Sequence

LOCUS       40924_49467        7428 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector YCplac22 5-1.2-FLuciferase, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7428)
  AUTHORS   Belmont BJ, Niles JC.
  TITLE     Inducible Control of Subcellular RNA Localization Using a Synthetic 
            Protein-RNA Aptamer Interaction
  JOURNAL   PLoS ONE 7 (10), E46868 (2012)
  PUBMED    23056498
REFERENCE   2  (bases 1 to 7428)
  AUTHORS   Belmont BJ, Niles JC.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-SEP-2012) Biological Engineering, Massachusetts 
            Institute of Technology, 77 Massachusetts Ave, Cambridge, MA 02139, 
            USA
REFERENCE   3  (bases 1 to 7428)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7428)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2012"; volume: "7"; issue: "10"; pages: "E46868"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (12-SEP-2012) Biological Engineering, Massachusetts Institute of 
            Technology, 77 Massachusetts Ave, Cambridge, MA 02139, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     17..33
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             678..2327
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(2632..2648)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(3407..4569)
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the
                     autonomously replicating sequence ARS1/ARS416"
     promoter        complement(5083..5184)
                     /label=TRP1 promoter
     promoter        5290..5394
                     /label=AmpR promoter
     CDS             5395..6252
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6426..7014
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    7302..7323
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        7338..7368
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7376..7392
                     /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     7400..7416
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"