Basic Vector Information
- Vector Name:
- pNC-Green0029-35S
- Antibiotic Resistance:
- Kanamycin
- Length:
- 6122 bp
- Type:
- Plant binary expression vector
- Replication origin:
- pSa ori
- Host:
- Plants
- Source/Author:
- Yan P, Zeng Y, Shen W, Tuo D
- Promoter:
- CaMV35S(short)
pNC-Green0029-35S 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 ℃.
pNC-Green0029-35S vector Sequence
LOCUS 62056_17825 6122 bp DNA circular SYN 04-SEP-2019 DEFINITION Plant binary expression vector pNC-Green0029-35S, complete sequence. ACCESSION MK896904 VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6122) AUTHORS Yan P, Zeng Y, Shen W, Tuo D, Li X, Zhou P. TITLE Direct Submission JOURNAL Submitted (07-MAY-2019) Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agriculture Sciences, Xueyuan Road, Haikou, Hainan 571101, China REFERENCE 2 (bases 1 to 6122) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Submitted (07-MAY-2019) Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agriculture Sciences, Xueyuan Road, Haikou, Hainan 571101, China" FEATURES Location/Qualifiers source 1..6122 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 184..529 /label=CaMV 35S promoter /note="strong constitutive promoter from cauliflower mosaic virus" promoter 725..827 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 858..1160 /codon_start=1 /label=ccdB /note="CcdB, a bacterial toxin that poisons DNA gyrase" /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" terminator 1251..1503 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" primer_bind complement(1521..1537) /label=SK primer /note="common sequencing primer, one of multiple similar variants" promoter complement(1574..1592) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(1613..1629) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(1637..1653) /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(1661..1692) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(1707..1728) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." misc_feature 1967..1991 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" rep_origin complement(2082..2670) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(2774..3586) /codon_start=1 /label=KanR /note="aminoglycoside phosphotransferase" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYGLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" rep_origin 3877..4312 /label=pSa ori /note="origin of replication from bacterial plasmid pSa" misc_feature 4445..4467 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA (truncated)" terminator complement(4491..4742) /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" CDS complement(4785..5576) /codon_start=1 /label=NeoR/KanR /note="aminoglycoside phosphotransferase" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERGRTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTQGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" promoter complement(5610..5789) /label=NOS promoter /note="nopaline synthase promoter" primer_bind 6036..6052 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 6059..6077 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 6103..6119 /label=KS primer /note="common sequencing primer, one of multiple similar variants"
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