Basic Vector Information
- Vector Name:
- pLeo1209
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5716 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Krawczyk K, Scheller L, Kim H, Fussenegger M.
- Promoter:
- U6
pLeo1209 vector Vector Map
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 ℃.
pLeo1209 vector Sequence
LOCUS 62056_13955 5716 bp DNA circular SYN 15-FEB-2020
DEFINITION Expression vector pLeo1209, complete sequence.
ACCESSION MN811119
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5716)
AUTHORS Krawczyk K, Scheller L, Kim H, Fussenegger M.
TITLE Rewiring of endogenous signaling pathways to genomic targets for
therapeutic cell reprogramming
JOURNAL Nat Commun 11 (1), 608 (2020)
PUBMED 32001704
REFERENCE 2 (bases 1 to 5716)
AUTHORS Scheller L, Krawczyk K.
TITLE Direct Submission
JOURNAL Submitted (09-DEC-2019) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel
4058, Switzerland
REFERENCE 3 (bases 1 to 5716)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Commun"; date: "2020"; volume: "11"; issue: "1"; pages: "608"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(09-DEC-2019) D-BSSE, ETH Zurich, Mattenstrasse 26, Basel 4058,
Switzerland"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..5716
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(579..1436)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(1437..1541)
/label=AmpR promoter
polyA_signal complement(1973..2197)
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
misc_RNA complement(2365..2383)
/label=MS2 stem loop
/note="stem loop that binds the bacteriophage MS2 coat
protein"
promoter complement(2429..2669)
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
CDS 3386..4084
/codon_start=1
/label=TagBFP
/note="monomeric blue fluorescent protein"
/translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
DLPSKLGHKLN"
CDS 4151..4747
/codon_start=1
/label=PuroR
/note="puromycin N-acetyltransferase"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
polyA_signal 4915..4970
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
rep_origin complement(join(5533..5716,1..405))
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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