pLeo1216 vector (V015391)

Basic Vector Information

Vector Name:
pLeo1216
Antibiotic Resistance:
Ampicillin
Length:
6347 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Scheller L, Schmollack M, Bertschi A, Mansouri M
Promoter:
SV40

pLeo1216 vector Vector Map

pLeo12166347 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300AmpRAmpR promoterDcuR BE2DcuR BE1DcuR BE2DcuR BE1minimal CMV promoterT7 promoterKozak+ATGSEAPbGH poly(A) signalf1 oriSV40 promoterNeoR/KanRSV40 poly(A) signalM13 revlac operatorlac promoterCAP binding siteori

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 ℃.

pLeo1216 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       62056_13960        6347 bp DNA     circular SYN 30-JUN-2020
DEFINITION  Cloning vector pLeo1216, complete sequence.
ACCESSION   MT267301
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6347)
  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 6347)
  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 6347)
  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..6347
                     /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    1029..1050
                     /label=DcuR BE2
                     /note="DcuR BE2"
     misc_feature    1069..1094
                     /label=DcuR BE1
                     /note="DcuR BE1"
     misc_feature    1108..1133
                     /label=DcuR BE2
                     /note="DcuR BE2"
     misc_feature    1152..1177
                     /label=DcuR BE1
                     /note="DcuR BE1"
     promoter        1191..1229
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        1274..1292
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1321..1326
                     /label=Kozak+ATG
                     /note="Kozak+ATG"
     CDS             1333..2829
                     /label=SEAP
                     /note="secreted alkaline phosphatase from human placenta"
     polyA_signal    2944..3168
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3214..3642
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3656..3985
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4052..4843
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    5020..5153
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5190..5206)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5214..5230)
                     /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(5238..5268)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5283..5304)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5592..6177)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"

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