Basic Vector Information
- Vector Name:
- pOBCol3.6-GFPtpz
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 9671 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells
- Promoter:
- Col1a1
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
pOBCol3.6-GFPtpz 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 ℃.
pOBCol3.6-GFPtpz vector Sequence
LOCUS 62056_18265 9671 bp DNA circular SYN 01-JAN-1980
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9671)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..9671
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 867..888
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 903..933
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 941..957
/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 965..981
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 1002..1020
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 1063..4693
/label=Col1a1
/note="Rat collagen type I alpha-1 promoter"
primer_bind 6294..6310
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
CDS 6383..7099
/codon_start=1
/label=Topaz YFP
/note="enhanced yellow variant of GFP"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMRQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
polyA_signal 7139..7363
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(7378..7394)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
promoter complement(7420..7438)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(7448..7464)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 7606..8061
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 8087..8191
/label=AmpR promoter
CDS complement(8683..9339)
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(9340..9442)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
rep_origin 9662..9671
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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