Basic Vector Information
- Vector Name:
- pMLBAD_GlycoLoopGFP
- Antibiotic Resistance:
- Trimethoprim
- Length:
- 7565 bp
- Type:
- Expression vector
- Replication origin:
- pBBR1 oriV
- Source/Author:
- Keys TG, Wetter M, Hang I, Rutschmann C, Russo S, Mally M, Steffen M, Zuppiger M, Muller F, Schneider J, Faridmoayer A, Lin CW, Aebi M.
- Promoter:
- araBAD
pMLBAD_GlycoLoopGFP 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 ℃.
pMLBAD_GlycoLoopGFP vector Sequence
LOCUS 40924_31095 7565 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pMLBAD_GlycoLoopGFP, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7565)
AUTHORS Keys TG, Wetter M, Hang I, Rutschmann C, Russo S, Mally M, Steffen
M, Zuppiger M, Muller F, Schneider J, Faridmoayer A, Lin CW, Aebi M.
TITLE A biosynthetic route for polysialylating proteins in Escherichia
coli
JOURNAL Metab. Eng. (2017) In press
PUBMED 29101090
REFERENCE 2 (bases 1 to 7565)
AUTHORS Keys TG.
TITLE Direct Submission
JOURNAL Submitted (21-NOV-2016) Institute of Microbiology, ETH Zurich,
Vladimir-Prelog-Weg 4, Zurich, ZH 8093, Switzerland
REFERENCE 3 (bases 1 to 7565)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7565)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Metab. Eng.
(2017) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(21-NOV-2016) Institute of Microbiology, ETH Zurich,
Vladimir-Prelog-Weg 4, Zurich, ZH 8093, Switzerland"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..7565
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 55..83
/label=Pc promoter
/note="class 1 integron promoter"
CDS 410..643
/codon_start=1
/label=TpR
/note="E. coli plasmid-associated dihydrofolate reductase"
/translation="MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGW
YCTKLTPEGYAVESESHPGSVQIYPVAALERVA"
CDS complement(872..1747)
/codon_start=1
/label=araC
/note="L-arabinose regulatory protein"
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
promoter 1774..2054
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
RBS 2063..2085
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 2101..2127
/codon_start=1
/label=9xHis
/note="9xHis affinity tag"
/translation="HHHHHHHHH"
CDS 2872..2895
/codon_start=1
/label=Strep-Tag II
/note="peptide that binds Strep-Tactin(R), an engineered
form
of streptavidin"
/translation="WSHPQFEK"
terminator 3132..3218
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3310..3337
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter complement(4192..4294)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS complement(4444..5463)
/codon_start=1
/gene="mob"
/product="Mob"
/function="required for plasmid mobilization"
/label=mob
/protein_id="ATN32199.1"
/translation="MAAYAIMRCKKLAKMGNVAASLKHAYRERETPNADASRTPENEHW
AASSTDEAMGRLRELLPEKRRKDAVLAVEYVMTASPEWWKSASQEQQAAFFEKAHKWLA
DKYGADRIVTASIHRDETSPHMTAFVVPLTQDGRLSAKEFIGNKAQMTRDQTTFAAAVA
DLGLQRGIEGSKARHTRIQAFYEALERPPVGHVTISPQAVEPRAYAPQGLAEKLGISKR
VETPEAVADRLTKAVRQGYEPALQAAAGAREMRKKADQAQETARDLRERLKPVLDALGP
LNRDMQAKAAAIIKAVGEKLLTEQREVQRQKQAQRQQERGRAHFPEKCHLTSKKPLLS"
gene complement(4444..5463)
/gene="mob"
/label=mob
rep_origin 5687..6456
/label=pBBR1 oriV
/note="replication origin of the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica; requires the pBBR1
Rep protein for replication"
CDS 6457..7116
/codon_start=1
/label=pBBR1 Rep
/note="replication protein for the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica"
/translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
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