Basic Vector Information
- Vector Name:
- pNPTS138-plasmid-pNPTS138
- Antibiotic Resistance:
- Kanamycin
- Length:
- 5361 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Ferro JA, Facincani AP, Tezza RD, Varani AM
- Promoter:
- sacB
pNPTS138-plasmid-pNPTS138 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 ℃.
pNPTS138-plasmid-pNPTS138 vector Sequence
LOCUS 62056_18190 5361 bp DNA circular SYN 17-JUL-2019
DEFINITION Cloning vector pNPTS138 plasmid pNPTS138, complete sequence.
ACCESSION MK533795
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5361)
AUTHORS Ferro JA, Facincani AP, Tezza RD, Varani AM, Ferreira H.
TITLE The nucleotide sequence of the plasmid pNPTS138
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5361)
AUTHORS Ferro JA, Facincani AP, Tezza RD, Varani AM, Ferreira H.
TITLE Direct Submission
JOURNAL Submitted (18-FEB-2019) Departamento de Tecnologia, Univ. Estadual
Paulista 'Julio de Mesquita Filho' - UNESP-Jaboticabal, Via de
Acesso Prof.Paulo Donato Castellane s/n, Jaboticabal, SP 14884-900,
Brazil
REFERENCE 3 (bases 1 to 5361)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(18-FEB-2019) Departamento de Tecnologia, Univ. Estadual Paulista
'Julio de Mesquita Filho' - UNESP-Jaboticabal, Via de Acesso
Prof.Paulo Donato Castellane s/n, Jaboticabal, SP 14884-900, Brazil"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
The plasmid was a gift from Prof. Lucy Shapiro, Department of
Developmental Biology, Stanford University School of Medicine
Beckman Center 279 Campus Drive, B300 Stanford, CA 94305.
FEATURES Location/Qualifiers
source 1..5361
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(158..174)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(182..198)
/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(206..236)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(251..272)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(333..922)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
oriT 1140..1249
/label=oriT
/note="incP origin of transfer"
CDS 1282..1650
/codon_start=1
/label=traJ
/note="oriT-recognizing protein"
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
rep_origin 1824..2275
/label=M13 ori
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS 2371..3183
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKFLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
promoter 3258..3685
/label=sacB promoter
/note="sacB promoter and control region"
CDS 3686..5104
/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"
primer_bind 5316..5332
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
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