Basic Vector Information
pTip-LNH2 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 ℃.
pTip-LNH2 vector Sequence
LOCUS 40924_43418 8193 bp DNA circular SYN 18-DEC-2018 DEFINITION Expression vector pTip-LNH2 DNA, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8193) AUTHORS Nakashima N, Tamura T. TITLE A novel system for expressing recombinant proteins over a wide temperature range from 4 to 35 degrees C JOURNAL Biotechnol. Bioeng. 86 (2), 136-148 (2004) PUBMED 15052633 REFERENCE 2 (bases 1 to 8193) AUTHORS Nakashima N, Tamura T. TITLE Direct Submission JOURNAL Submitted (18-OCT-2002) Nobutaka Nakashima, National Institute of Advanced Industrial Science and Technology; Tsukisamu-Higashi 2-17-2-1, Sapporo, Toyohira-ku 062-8517, Japan (E-mail:n-nakashima@aist.go.jp, Tel:81-11-857-8937) REFERENCE 3 (bases 1 to 8193) TITLE Direct Submission REFERENCE 4 (bases 1 to 8193) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol. Bioeng."; date: "2004"; volume: "86"; issue: "2"; pages: "136-148" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (18-OCT-2002) Nobutaka Nakashima, National Institute of Advanced Industrial Science and Technology"; volume: " Tsukisamu-Higashi 2-17-2-1, Sapporo, Toyohira-ku 062-8517, Japan (E-mail:n-nakashima@aist.go.jp, Tel"; pages: "81-11-857-8937" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..8193 /mol_type="other DNA" /organism="synthetic DNA construct" CDS 170..976 /gene="tsnR" /label=tsnR /note="23S rRNA (adenosine(1067)-2'-O)-methyltransferase from Streptomyces azureus. Accession#: P18644" regulatory 1145..1324 /gene="Tuf1" /regulatory_class="promoter" gene 1145..1324 /gene="Tuf1" /label=Tuf1 CDS 1325..2512 /label=TcR /note="tetracycline efflux protein" regulatory 2728..2903 /gene="ThcA" /regulatory_class="promoter" gene 2728..2903 /gene="ThcA" /label=ThcA CDS 2904..3662 /gene="tipA" /label=tipA /note="HTH-type transcriptional activator TipA from Streptomyces lividans. Accession#: P0A4T9" promoter 3843..3947 /label=AmpR promoter CDS 3948..4805 /label=AmpR /note="beta-lactamase" rep_origin 4979..5567 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" regulatory complement(5818..5998) /gene="ThcA" /regulatory_class="terminator" gene complement(5818..5998) /gene="ThcA" /label=ThcA CDS complement(6063..6080) /label=6xHis /note="6xHis affinity tag" RBS complement(6094..6116) /label=RBS /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" regulatory complement(6130..6253) /gene="TipA" /regulatory_class="promoter" CDS 6792..7679 /codon_start=1 /gene="RepA" /label=RepA /protein_id="BAD11248.1" /translation="MKSATGGVLSDRQWADAVLLPHRPFATNNLQRGQYRMSRDDALAM RYVEHSPHALLGSIVIDCDHVDAAMRAFEQPSDHPAPNWVAQSPSGRAHIGWWLGPNHV CRTDSARLTPLRYAHRIETGLKISVGGDFAYGGQLTKNPIHPDWETIYGPATPYTLRQL ATIHTPRQMPRRPDRAVGLGRNVTMFDATRRWAYPQWWQHRNGTGRDWDHLVLQHCHAV NTEFTTPLPFTEVRATAQSISKWIWRNFTEEQYRARQAHLGQKGGKATTLAKQEAVRNN ARKYDEHTMREAII" gene 6792..7679 /gene="RepA" /label=RepA CDS 7679..7963 /codon_start=1 /gene="RepB" /label=RepB /protein_id="BAD11249.1" /translation="MGGAKNPVRRKMTAAAAAEKFGASTRTIQRLFAEPRDDYLGRAKA RRDKAVELRKQGLKYREIAEAMELSTGIVGRLLHDARRHGEISAEDLSA" gene 7679..7963 /gene="RepB" /label=RepB
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