Basic Vector Information
- Vector Name:
- pTY2b
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4804 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Ohnishi N, Mukherjee B, Tsujikawa T, Yanase M, Nakano H, Moroney JV, Fukuzawa H.
- Promoter:
- T3
pTY2b 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 ℃.
pTY2b vector Sequence
LOCUS 40924_44574 4804 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pTY2b DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4804)
AUTHORS Ohnishi N, Mukherjee B, Tsujikawa T, Yanase M, Nakano H, Moroney JV,
Fukuzawa H.
TITLE Expression of a low CO-inducible protein, LCI1, increases inorganic
carbon uptake in the green alga Chlamydomonas reinhardtii
JOURNAL Plant Cell 22 (9), 3105-3117 (2010)
PUBMED 20870960
REFERENCE 2 (bases 1 to 4804)
AUTHORS Tsujikawa T, Fukuzawa H.
TITLE Direct Submission
JOURNAL Submitted (11-JUL-2008) Contact:Hideya Fukuzawa Kyoto University,
Graduate School of Biostudies; Kitashirakawa Oiwake-cho, Kyoto,
Kyoto 606-8502, Japan URL :http://chlamy.pmb.lif.kyoto-u.ac.jp/
REFERENCE 3 (bases 1 to 4804)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4804)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plant
Cell"; date: "2010"; volume: "22"; issue: "9"; pages: "3105-3117"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(11-JUL-2008) Contact:Hideya Fukuzawa Kyoto University, Graduate
School of Biostudies"; volume: " Kitashirakawa Oiwake-cho, Kyoto,
Kyoto 606-8502, Japan URL :http"; pages:
"//chlamy.pmb.lif.kyoto-u.ac.jp"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4804
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 24..128
/label=AmpR promoter
CDS 129..986
/label=AmpR
/note="beta-lactamase"
rep_origin 1160..1748
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2036..2057
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2072..2102
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2110..2126
/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)."
primer_bind 2134..2150
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 2171..2189
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
regulatory 2208..2291
/note="includes enhancer region (-231 to -200 and -77 to
-25) of Chlamydomonas reinhardtii NAD(P)H nitrate reductase
protein (Nit1) gene"
/regulatory_class="enhancer"
regulatory 2292..2375
/note="includes enhancer region (-231 to -200 and -77 to
-25) of Chlamydomonas reinhardtii NAD(P)H nitrate reductase
protein (Nit1) gene"
/regulatory_class="enhancer"
regulatory 2376..2411
/note="includes promoter region (-36 to -1) of
Chlamydomonas reinhardtii beta2-tubulin (tubB2) gene"
/regulatory_class="promoter"
misc_feature 2418..2612
/note="includes intron 1 of Chlamydomonas reinhardtii
nuclear gene rbcS2 for ribulose bisphosphate
carboxylase/oxygenase small subunit"
primer_bind 2632..2648
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 2680..2863
/note="3'UTR region of Chlamydomonas reinhardtii nuclear
gene rbcS2 for ribulose bisphosphate carboxylase oxygenase
small subunit"
primer_bind complement(2945..2961)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
regulatory 2974..3188
/note="includes promoter region of Chlamydomonas
reinhardtii nuclear gene rbcS2 for ribulose bisphosphate
carboxylase/oxygenase small subunit"
/regulatory_class="promoter"
misc_feature 3188..3333
/note="includes intron 1 from Chlamydomonas reinhardtii
rbcS2 gene"
CDS join(3334..3501,3647..3859)
/codon_start=1
/gene="ble"
/product="bleomycin resistance protein"
/label=ble
/note="derived from Streptoalloteichus hindustanus; fusion
zeocin binding protein; confers resistance to zeocin"
/protein_id="BAI39424.1"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPW
GREFALRDPAGNCVHFVAEEQD"
misc_feature 3334..3501
/gene="ble"
/note="includes a fragment of Streptoalloteichus
hindustanus Ble gene"
gene 3334..3501
/gene="ble"
/label=ble
misc_feature 3502..3646
/note="includes intron 1 of Chlamydomonas reinhardtii rbcS2
gene"
misc_feature 3647..3859
/gene="ble"
/note="includes a fragment of Streptoalloteichus
hindustanus Ble gene and stop codon"
gene 3647..3859
/gene="ble"
/label=ble
promoter complement(4161..4179)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4189..4205)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 4347..4802
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
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