Basic Vector Information
p1enVT-lacI 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 ℃.
p1enVT-lacI vector Sequence
LOCUS 40924_2458 5431 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector p1enVT-lacI, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5431)
AUTHORS Boehm CR, Grant PK, Haseloff J.
TITLE Programmed emergence of a domain of gene expression in a bacterial
population
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 5431)
AUTHORS Boehm CR.
TITLE Direct Submission
JOURNAL Submitted (12-OCT-2016) Plant Sciences, University of Cambridge,
Downing Street, Cambridge, Cambridgeshire CB2 3EA, United Kingdom
REFERENCE 3 (bases 1 to 5431)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5431)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(12-OCT-2016) Plant Sciences, University of Cambridge, Downing
Street, Cambridge, Cambridgeshire CB2 3EA, United Kingdom"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..5431
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(49..637)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 825..843
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 875..897
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 909..1625
/label=Venus
/note="yellow fluorescent protein (YFP) with fast and
efficient maturation (Nagai et al., 2002)"
terminator 1648..1719
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 1735..1762
/label=T7Te terminator
/note="phage T7 early transcription terminator"
regulatory 1769..1803
/label=PJ23101*
/note="PJ23101*"
/regulatory_class="promoter"
RBS 1812..1823
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 1839..2918
/label=lacI
/note="lac repressor"
CDS 2919..2951
/label=ssrA tag (LVA)
/note="C-terminal peptide that mediates degradation in
bacteria through the ClpXP and ClpAP proteases (McGinness
et al., 2006)"
terminator complement(3002..3045)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3086..3113
/label=T7Te terminator
/note="phage T7 early transcription terminator"
misc_feature 3120..3140
/label=BioBrick suffix
/note="universal suffix for all parts"
CDS complement(3166..4023)
/label=AmpR
/note="beta-lactamase"
promoter complement(4024..4128)
/label=AmpR promoter
terminator complement(4218..4261)
/label=bacterial terminator
/note="putative bacterial transcription terminator"
misc_feature 4264..4285
/label=BioBrick prefix
/note="BioBrick prefix for parts that do not start with
'ATG'"
terminator complement(4292..4319)
/label=T7Te terminator
/note="phage T7 early transcription terminator"
terminator complement(4335..4406)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(4429..5145)
/label=mTurquoise2
/note="enhanced monomeric variant of CFP (Goedhart et al.,
2012)"
regulatory complement(5146..5163)
/label=BBa_B0034
/note="BBa_B0034"
/regulatory_class="ribosome_binding_site"
RBS 5152..5163
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
regulatory complement(5172..5206)
/label=pJ23101
/note="pJ23101"
/regulatory_class="promoter"
misc_feature 5207..5227
/label=BioBrick suffix
/note="universal suffix for all parts"
terminator 5228..5285
/label=his operon terminator
/note="This putative transcriptin terminator from the E.
coli his operon has a 2-bp deletion introduced during
synthesis. Its efficiency has not been determined."
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