Basic Vector Information
pTpPuc3 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 ℃.
pTpPuc3 vector Sequence
LOCUS 40924_43823 7796 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pTpPuc3, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7796)
AUTHORS Karas BJ, Diner RE, Lefebvre SC, Weyman PD.
TITLE Designer Diatom episomes Delivered by Bacterial Conjugation
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7796)
AUTHORS Karas BJ, Diner RE, Lefebvre SC, Weyman PD.
TITLE Direct Submission
JOURNAL Submitted (03-FEB-2015) Synthetic Biology and Bioenergy Group, J.
Craig Venter Institute, 4120 Capricorn Lane, La Jolla, CA 92037, USA
REFERENCE 3 (bases 1 to 7796)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7796)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(03-FEB-2015) Synthetic Biology and Bioenergy Group, J. Craig Venter
Institute, 4120 Capricorn Lane, La Jolla, CA 92037, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..7796
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(310..678)
/codon_start=1
/label=traJ
/note="oriT-recognizing protein"
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
oriT complement(711..820)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
primer_bind 1098..1114
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS 2096..2662
/codon_start=1
/label=NrsR
/note="nourseothricin acetyltransferase"
/translation="MTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGFT
LREVPVDPPLTKVFPDDESDDESDAGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNRR
LTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFTL
CGLDTALYDGTASDGEQALYMSMPCP"
misc_feature 3185..3688
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 3689..3876
/label=HIS3 promoter
CDS 3877..4533
/codon_start=1
/label=HIS3
/note="imidazoleglycerol-phosphate dehydratase, required
for histidine biosynthesis"
/translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
primer_bind complement(4605..4621)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4629..4645)
/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)."
promoter complement(4653..4683)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4698..4719)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5007..5595)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS 6443..7255
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
promoter complement(7597..7701)
/label=AmpR promoter
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