Basic Vector Information
- Vector Name:
- pEGC
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 5829 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Becker K, Meyer A, Roberts TM, Panke S.
pEGC 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 ℃.
pEGC vector Sequence
LOCUS 62056_8945 5829 bp DNA circular SYN 28-JUL-2021
DEFINITION Cloning vector pEGC, complete sequence.
ACCESSION MZ395121
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5829)
AUTHORS Becker K, Meyer A, Roberts TM, Panke S.
TITLE Plasmid replication based on the T7 origin of replication requires a
T7 RNAP variant and inactivation of ribonuclease H
JOURNAL Nucleic Acids Res (2021) In press
PUBMED 34255845
REFERENCE 2 (bases 1 to 5829)
AUTHORS Becker K.
TITLE Direct Submission
JOURNAL Submitted (10-JUN-2021) D-BSSE, ETHZ, Mattenstrasse 26, Basel, BS
4058, Switzerland
REFERENCE 3 (bases 1 to 5829)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res (2021) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(10-JUN-2021) D-BSSE, ETHZ, Mattenstrasse 26, Basel, BS 4058,
Switzerland"
FEATURES Location/Qualifiers
source 1..5829
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 17..605
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(791..933)
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(1038..1226)
/codon_start=1
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
/translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
protein_bind complement(2001..2022)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(2038..3117)
/codon_start=1
/label=lacI
/note="lac repressor"
/translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
promoter complement(3118..3195)
/label=lacI promoter
promoter 3504..3522
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 3523..3547
/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 3562..3584
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 3592..4305
/codon_start=1
/label=superfolder GFP
/note="GFP variant that folds robustly even when fused to
poorly folded proteins (Nager et al., 2011)"
/translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
CDS 4312..4329
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
CDS 4339..4356
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
terminator 4423..4470
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 4507..4962
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(5054..5710)
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(5711..5813)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
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