p1g3VT-lacI vector (V009776)

Basic Vector Information

Vector Name:
p1g3VT-lacI
Antibiotic Resistance:
Ampicillin
Length:
5413 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Boehm CR, Grant PK, Haseloff J.

p1g3VT-lacI vector Vector Map

p1g3VT-lacI5413 bp60012001800240030003600420048005400oriPT7(-3G)BBa_B0034VenusrrnB T1 terminatorT7Te terminatorPJ23101*RBSlacIssrA tag (LVA)rrnB T1 terminatorT7Te terminatorBioBrick suffixAmpRAmpR promoterbacterial terminatorBioBrick prefixT7Te terminatorrrnB T1 terminatormTurquoise2BBa_B0034pJ23101BioBrick suffixhis operon terminator

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 ℃.

p1g3VT-lacI vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_2463        5413 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector p1g3VT-lacI, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5413)
  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  JOURNAL   Unpublished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 5413)
  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 5413)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5413)
  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            "Unpublished"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..5413
                     /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"
     regulatory      818..863
                     /label=PT7(-3G)
                     /note="PT7(-3G)"
                     /regulatory_class="promoter"
     regulatory      872..883
                     /label=BBa_B0034
                     /note="BBa_B0034"
                     /regulatory_class="ribosome_binding_site"
     RBS             872..883
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             890..1606
                     /label=Venus
                     /note="yellow fluorescent protein (YFP) with fast and
                     efficient maturation (Nagai et al., 2002)"
     terminator      1629..1700
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1716..1743
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     regulatory      1750..1784
                     /label=PJ23101*
                     /note="PJ23101*"
                     /regulatory_class="promoter"
     RBS             1793..1804
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1820..2899
                     /label=lacI
                     /note="lac repressor"
     CDS             2900..2932
                     /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(2983..3026)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3067..3094
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    3101..3121
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     CDS             complement(3147..4004)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(4005..4109)
                     /label=AmpR promoter
     terminator      complement(4199..4242)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    4245..4266
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with
                     'ATG'"
     terminator      complement(4273..4300)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(4316..4387)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(4410..5126)
                     /label=mTurquoise2
                     /note="enhanced monomeric variant of CFP (Goedhart et al., 
                     2012)"
     regulatory      complement(5127..5144)
                     /label=BBa_B0034
                     /note="BBa_B0034"
                     /regulatory_class="ribosome_binding_site"
     RBS             5133..5144
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     regulatory      complement(5153..5187)
                     /label=pJ23101
                     /note="pJ23101"
                     /regulatory_class="promoter"
     misc_feature    5188..5208
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      5209..5266
                     /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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