Basic Vector Information
- Vector Name:
- pLW18
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9064 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Delaney NF, Kaczmarek ME, Ward LM, Swanson PK, Lee MC, Marx CJ.
- Promoter:
- sacB
pLW18 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 ℃.
pLW18 vector Sequence
LOCUS 40924_29311 9064 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pLW18, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9064)
AUTHORS Delaney NF, Kaczmarek ME, Ward LM, Swanson PK, Lee MC, Marx CJ.
TITLE Development of an Optimized Medium, Strain and High-Throughput
Culturing Methods for Methylobacterium extorquens
JOURNAL PLoS ONE 8 (4), E62957 (2013)
PUBMED 23646164
REFERENCE 2 (bases 1 to 9064)
AUTHORS Delaney NF.
TITLE Direct Submission
JOURNAL Submitted (25-MAR-2013) Organismic and Evolutionary Biology, Harvard
University, 16 Divinity Ave., Cambridge, MA 02138, USA
REFERENCE 3 (bases 1 to 9064)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9064)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE";
date: "2013"; volume: "8"; issue: "4"; pages: "E62957"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(25-MAR-2013) Organismic and Evolutionary Biology, Harvard
University, 16 Divinity Ave., Cambridge, MA 02138, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..9064
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(4..861)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(862..966)
/label=AmpR promoter
misc_feature 997..1548
/label=region upstream of celA gene
/note="region upstream of celA gene"
misc_feature 1549..2075
/label=region downstream of celC gene
/note="region downstream of celC gene"
CDS complement(2679..3047)
/codon_start=1
/label=traJ
/note="oriT-recognizing protein"
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
oriT complement(3080..3189)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
promoter 3826..3854
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
CDS 3902..5089
/codon_start=1
/label=TcR
/note="tetracycline efflux protein"
/translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
CDS complement(5273..5929)
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS complement(6030..7448)
/codon_start=1
/label=SacB
/note="secreted levansucrase that renders bacterial growth
sensitive to sucrose"
/translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
VKDSILEQGQLTVNK"
promoter complement(7449..7894)
/label=sacB promoter
/note="sacB promoter and control region"
rep_origin complement(8306..8894)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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