pLeo1254 vector (V016159)

MSDS

Basic Vector Information

Vector Name:: pLeo1254

Antibiotic Resistance:: Ampicillin

Length:: 5866 bp

Type:: Cloning vector

Replication origin:: ori

Source/Author:: Scheller L, Schmollack M, Bertschi A, Mansouri M

pLeo1254 vector Vector Map

View Map in NovoBuilder

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 ℃.

pLeo1254 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       62056_14010        5866 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pLeo1254, complete sequence.
ACCESSION   MT267311
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5866)
  AUTHORS   Scheller L, Schmollack M, Bertschi A, Mansouri M, Saxena P, 
            Fussenegger M.
  TITLE     Phosphoregulated orthogonal signal transduction in mammalian cells
  JOURNAL   Nat Commun 11 (1), 3085 (2020)
  PUBMED    32555187
REFERENCE   2  (bases 1 to 5866)
  AUTHORS   Scheller L.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-MAR-2020) STI IBI-STI LPDI, EPFL, AI 3123 (Batiment 
            AI), Station 19, Lausanne 1015, Switzerland
REFERENCE   3  (bases 1 to 5866)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun"; date: "2020"; volume: "11"; issue: "1"; pages: "3085"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-MAR-2020) STI IBI-STI LPDI, EPFL, AI 3123 (Batiment AI), Station
            19, Lausanne 1015, Switzerland"
FEATURES             Location/Qualifiers
     source          1..5866
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(4..861)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(862..966)
                     /label=AmpR promoter
     misc_feature    1035..1238
                     /label=SV40 early promoter
                     /note="SV40 early promoter"
     promoter        1288..1306
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1338..2426
                     /codon_start=1
                     /transl_table=11
                     /product="C-ORK"
                     /label=C-ORK
                     /protein_id="QKE44350.1"
                     /translation="MTSKTEVRKLMQRLDGLVNYADALRERSHEFMNKLHVILGLLHLK
                     SYKQLEDYILKTANNYQEEIGSLLGKIKSPVIAGFLISKINRATDLGHTLILNSESQLP
                     DSGSEDQVATLITTLGNLIENALEALGPEPGGEISVTLHYRHGWLHCEVNDDGPGIAPD
                     KIDHIFDKGVSTKGSERGVGLALVKQQVENLGGSIAVESEPGIFTQFFVQIPWDGERSN
                     RASASGGGGSASGSQVQLVESGGGLVQAGGSLRLSCTASGRTGTIYSMAWFRQAPGKER
                     EFLATVGWSSGITYYMDSVKGRFTISRDKGKNTVYLQMDSLKPEDTAVYYCTATRAYSV
                     GYDYWGQGTQVTVSSASGSTGV"
     misc_feature    1365..1367
                     /label=leu330
                     /note="leu330"
     misc_feature    1377..1379
                     /label=leucin 334
                     /note="leucin 334"
     misc_feature    1389..1391
                     /label=V338
                     /note="V338"
     misc_feature    1392..1394
                     /label=asn339
                     /note="asn339"
     misc_feature    1395..1412
                     /label=alpha helix
                     /note="alpha helix"
     misc_feature    1413..2006
                     /label=Kinase domain 346-538
                     /note="Kinase domain 346-538"
     misc_feature    2034..2402
                     /label=aCaffVHH
                     /note="aCaffVHH"
     misc_feature    2121..2144
                     /label=CDR1
                     /note="CDR1"
     misc_feature    2193..2216
                     /label=CDR2
                     /note="CDR2"
     misc_feature    2334..2369
                     /label=CDR3
                     /note="CDR3"
     polyA_signal    2463..2687
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2733..3161
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3175..3504
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3571..4362
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    4539..4672
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4709..4725)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4733..4749)
                     /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)."
     promoter        complement(4757..4787)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4802..4823)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5111..5696)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"

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