Basic Vector Information
- Vector Name:
- pNIC-ZB
- Antibiotic Resistance:
- Kanamycin
- Length:
- 7446 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Savitsky P, Burgess-Brown N, Gileadi O.
- Copy Number:
- High copy number
- Promoter:
- sacB
pNIC-ZB 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 ℃.
pNIC-ZB vector Sequence
LOCUS 40924_33247 7446 bp DNA circular SYN 18-DEC-2018 DEFINITION Expression vector pNIC-ZB, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7446) AUTHORS Savitsky P, Burgess-Brown N, Gileadi O. TITLE Direct Submission JOURNAL Submitted (12-JAN-2010) Structural Genomics Consortium, University of Oxford, Old Road Campus Research Building, Roosevelt Drive, Oxford OX3 7DQ, UK REFERENCE 2 (bases 1 to 7446) TITLE Direct Submission REFERENCE 3 (bases 1 to 7446) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Submitted (12-JAN-2010) Structural Genomics Consortium, University of Oxford, Old Road Campus Research Building, Roosevelt Drive, Oxford OX3 7DQ, UK" COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..7446 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 1..19 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 20..44 /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)." RBS 59..81 /label=RBS /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 92..109 /codon_start=1 /label=6xHis /note="6xHis affinity tag" /translation="HHHHHH" CDS 119..289 /codon_start=1 /label=Z-basic tag /note="Z domain of staphylococcal protein A, engineered to contain multiple positive charges (Hedhammar and Hober, 2007)" /translation="VDNKFNKERRRARREIRHLPNLNREQRRAFIRSLRDDPSQSANLL AEAKKLNDAQPK" CDS 296..316 /codon_start=1 /label=TEV site /note="tobacco etch virus (TEV) protease recognition and cleavage site" /translation="ENLYFQS" promoter 332..777 /label=sacB promoter /note="sacB promoter and control region" CDS 778..2196 /codon_start=1 /label=SacB /note="secreted levansucrase that renders bacterial growth sensitive to sucrose" /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH ITRHDMLQIPEQQKNEKYKVPEFDSSTIKNISSAKGLDVWDSWPLQNTDGTVANYHGYH IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV VKDSILEQGQLTVNK" misc_feature complement(2245..2259) /label=3'-LIC /note="3'-LIC" CDS 2307..2324 /codon_start=1 /label=6xHis /note="6xHis affinity tag" /translation="HHHHHH" terminator 2391..2438 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 2475..2930 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" CDS complement(3026..3838) /codon_start=1 /label=KanR /note="aminoglycoside phosphotransferase" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" rep_origin 3960..4548 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature complement(4734..4876) /label=bom /note="basis of mobility region from pBR322" CDS complement(4981..5169) /codon_start=1 /label=rop /note="Rop protein, which maintains plasmids at low copy number" /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" protein_bind complement(5944..5965) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." CDS complement(5981..7060) /codon_start=1 /label=lacI /note="lac repressor" /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" promoter complement(7061..7138) /label=lacI promoter
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