Basic Vector Information
- Vector Name:
- pMSW107
- Antibiotic Resistance:
- Kanamycin
- Length:
- 10933 bp
- Type:
- Cloning vector
- Replication origin:
- pBBR1 oriV
- Source/Author:
- Wecker MS, Beaton S, Chado R, Ghirardi ML.
- Promoter:
- T3
pMSW107 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 ℃.
pMSW107 vector Sequence
LOCUS 40924_32425 10933 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pMSW107, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10933)
AUTHORS Wecker MS, Beaton S, Chado R, Ghirardi ML.
TITLE Development of a Rhodobacter capsulatus self-reporting model system
for optimizing light-dependent, [FeFe]-hydrogenase-driven H
production
JOURNAL Biotechnol. Bioeng. (2016) In press
PUBMED 27531314
REFERENCE 2 (bases 1 to 10933)
AUTHORS Wecker MS.
TITLE Direct Submission
JOURNAL Submitted (26-MAY-2016) Molecular Biology, GeneBiologics, 1047
Cherryvale Road, Boulder, CO 80303, USA
REFERENCE 3 (bases 1 to 10933)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10933)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol.
Bioeng. (2016) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(26-MAY-2016) Molecular Biology, GeneBiologics, 1047 Cherryvale
Road, Boulder, CO 80303, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10933
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 458..1249
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
protein_bind 1657..1678
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 1693..1723
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1731..1747
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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 1755..1771
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 1792..1810
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
CDS complement(1832..2548)
/label=EmGFP
/note="Emerald GFP"
misc_feature complement(2549..2770)
/gene="HupSL-emGFP"
/note="HupSL remnant; acts as signal sequence for the
downstream emGFP"
regulatory complement(2771..3299)
/regulatory_class="promoter"
primer_bind 3306..3322
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3356..3372)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(3405..3423)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3433..3449)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
regulatory 3881..4491
/note="fru promoter; derived from Rhodobacter capsulatus"
/regulatory_class="promoter"
CDS 4492..5544
/codon_start=1
/gene="hydE"
/product="HydE"
/label=hydE
/note="derived from Clostridium acetobutylicum strain DSM
1731; codon-optimized for Rhodobacter capsulatus"
/protein_id="AOF40442.1"
/translation="MDNIIKLINKAEVTHDLTKDELVTLLKDDTHNEEIYKAADRVREK
YVGEEVHLRGLIEFSNICKRNCMYCGLRRDNKNIKRYRLEPDEIIHLAKSAKNYGYQTV
VLQSGEDDYYTVEKMKYIVSEIKKLNMAITLSIGEKTFEEYEEYRKSGADRYLIRIETT
DKELYEKLDPKMSHENRINCLKNLRKLGYEVGSGCLVGLPNQTIESLADDILFFKEIDA
DMIGVGPFIPNEDTPLGEEKGGEFFMSVKVTALIRLLLPDINIPATTAMESLYPNGRSI
ALTSGANVVMPNVTEGEYRKLYALYPGKICVNDTPGHCRQCISLKINKINRKVSATKGF
RKKSYKESIG"
gene 4492..5544
/gene="hydE"
/label=hydE
CDS 5558..6793
/codon_start=1
/gene="hydF"
/product="HydF"
/label=hydF
/note="derived from Clostridium acetobutylicum strain DSM
1731; codon-optimized for Rhodobacter capsulatus"
/protein_id="AOF40443.1"
/translation="MNELNSTPKGERLHIALFGKTNVGKSSVINALTSQEIALVSNVKG
TTTDPVYKAMELLPLGPVMLIDTAGLDDISDLGELRRGKTLEVLSKTDVAILVFDVESG
ITEYDKNIYSLLLEKKIPLIGVLNKIDKKDYKLEDYTSQFKIPIVPISALNNKGINNLK
DELIRLAPENDDKFKIVGDLLSPGDIAVLVTPIDKAAPKGRLILPQQQTIRDILESDAI
AMVTKEFELRETLDSLRKKPKIVITDSQVFLKVAADTPKDILMTSFSILMARHKGDLIE
LARGARAIEDLKDGDKILIAEACTHHRQSDDIGKVKIPRWLRQKTGKKLEFDFSSGFSF
PPNIEDYALIVHCAGCMLNRRSMLHRIESSVKKQIPIVNYGVLIAYVQGILPRALKPFP
YADRIFNQSSRN"
gene 5558..6793
/gene="hydF"
/label=hydF
CDS 6807..8225
/codon_start=1
/gene="hydG"
/product="HydG"
/label=hydG
/note="derived from Clostridium acetobutylicum strain DSM
1731; codon-optimized for Rhodobacter capsulatus"
/protein_id="AOF40444.1"
/translation="MYNVKSKVATEFISDEEIIDSLEYAKQNKSNRELIDSIIEKAKEC
KGLTHRDAAVLLECDLEDENEKMFKLAREIKQKFYGNRIVMFAPLYLSNYCVNGCVYCP
YHHKNKHIARKKLSQEDVKRETIALQDMGHKRLALEAGEDPVNNPIEYILDCIKTIYSI
KHKNGAIRRVNVNIAATTVENYKKLKDAGIGTYILFQETYNKKSYEELHPTGPKHDYAY
HTEAMDRAMEGGIDDVGIGVLFGLNMYKYDFVGLLMHAEHLEAAMGVGPHTISVPRIRP
ADDIDPENFSNAISDEIFEKIVAIIRIAVPYTGMIVSTRESKKTRERVLELGISQISGG
SSTSVGGYVESEPEEDNSSQFEVNDNRTLDEIVNWLLEMNYIPSFCTACYREGRTGDRF
MSLVKSGQIANCCQPNALMTLKEYLEDYASSNTQKNGEALIASEVEKIPNEKVKSIVKK
HLTELKEGQRDFRF"
gene 6807..8225
/gene="hydG"
/label=hydG
CDS complement(8482..9141)
/label=pBBR1 Rep
/note="replication protein for the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica"
rep_origin complement(9142..9911)
/direction=LEFT
/label=pBBR1 oriV
/note="replication origin of the broad-host-range plasmid
pBBR1 from Bordetella bronchiseptica; requires the pBBR1
Rep protein for replication"
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