Basic Vector Information
- Vector Name:
- MiniTn-Puro-JCVI-1.7
- Antibiotic Resistance:
- Kanamycin
- Length:
- 13902 bp
- Type:
- Tri-shuttle cloning vector
- Replication origin:
- ori2
- Host:
- Yeast
- Source/Author:
- Benders GA, Noskov VN, Denisova EA, Lartigue C, Gibson DG, Assad-Garcia N, Chuang RY, Carrera W, Moodie M, Algire MA, Phan Q, Alperovich N, Vashee S, Merryman C, Venter JC, Smith HO, Glass JI, Hutchis
- Promoter:
- HIS3
MiniTn-Puro-JCVI-1.7 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 ℃.
MiniTn-Puro-JCVI-1.7 vector Sequence
LOCUS 40924_1989 13902 bp DNA circular SYN 18-DEC-2018
DEFINITION Tri-shuttle cloning vector MiniTn-Puro-JCVI-1.7, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13902)
AUTHORS Benders GA, Noskov VN, Denisova EA, Lartigue C, Gibson DG,
Assad-Garcia N, Chuang RY, Carrera W, Moodie M, Algire MA, Phan Q,
Alperovich N, Vashee S, Merryman C, Venter JC, Smith HO, Glass JI,
Hutchison CA III.
TITLE Cloning whole bacterial genomes in yeast
JOURNAL Nucleic Acids Res. 38 (8), 2558-2569 (2010)
PUBMED 20211840
REFERENCE 2 (bases 1 to 13902)
AUTHORS Gibson DG, Benders GA, Lartigue C, Glass JI.
TITLE Direct Submission
JOURNAL Submitted (02-FEB-2010) Synthetic Biology, J. Craig Venter
Institute, 10355 Science Center Dr., San Diego, CA 92121, USA
REFERENCE 3 (bases 1 to 13902)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 13902)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res."; date: "2010"; volume: "38"; issue: "8"; pages:
"2558-2569"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(02-FEB-2010) Synthetic Biology, J. Craig Venter Institute, 10355
Science Center Dr., San Diego, CA 92121, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..13902
/mol_type="other DNA"
/organism="synthetic DNA construct"
mobile_element complement(1..26)
/mobile_element_type="insertion sequence:IS256"
/label=insertion sequence:IS256
/note="inner inverted repeat"
promoter complement(45..63)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(84..100)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(108..124)
/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(132..162)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(177..198)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(486..1074)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1248..2105)
/label=AmpR
/note="beta-lactamase"
promoter complement(2106..2210)
/label=AmpR promoter
rep_origin complement(2236..2691)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 2833..2849
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 2856..2874
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS complement(2895..4067)
/codon_start=1
/product="IS256 transposase"
/label=IS256 transposase
/protein_id="ADE19332.1"
/translation="MTQVHFTLKSEEIQSIIEYSVKDDVSKNILTTVFNQLMENQRTEY
IQAKEYERTENRQSQRNGYYERSFTTRVGTLELKVPRTRDGHFSPTVFERYQRNEKALM
ASMLEMYVSGVSTRKVSKIVEELCGKSVSKSFVSSLTEQLEPMVNEWQNRLLSEKNYPY
LMTDVLYIKVREENRVLSKSCHIAIGITKDGDREIIGFMIQSGESEETWTTFFEYLKER
GLQGTELVISDAHKGLVSAIRKSFTNVSWQRCQVHFLRNIFTTIPKKNSKSFREAVKGI
FKFTDINLAREAKNRLIHDYIDQPKYSKACASLDDGFEDAFQYTVQGNSHNRLKSTNLI
ERLNQEVRRREKIIRIFPNQTSANRLIGAVLMDLHDEWIYSSRKYINFDK"
mobile_element 4173..4197
/mobile_element_type="insertion sequence:IS256"
/label=insertion sequence:IS256
/note="outer inverted repeat"
protein_bind 4347..4380
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
terminator 4402..4431
/label=T3Te terminator
/note="phage T3 early transcription terminator"
rep_origin 4462..5091
/label=oriV
/note="incP origin of replication"
rep_origin 5165..5384
/label=ori2
/note="secondary origin of replication for the bacterial F
plasmid; also known as oriS"
CDS 5475..6227
/label=repE
/note="replication initiation protein for the bacterial F
plasmid"
misc_feature 6233..6483
/label=incC
/note="incompatibility region of the bacterial F plasmid"
CDS 6809..7981
/label=sopA
/note="partitioning protein for the bacterial F plasmid"
CDS 7984..8952
/label=sopB
/note="partitioning protein for the bacterial F plasmid"
misc_feature 9028..9501
/label=sopC
/note="centromere-like partitioning region of the bacterial
F plasmid"
CDS complement(9775..10587)
/label=KanR
/note="aminoglycoside phosphotransferase"
promoter complement(10588..10690)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
misc_feature complement(10712..11110)
/label=cos
/note="lambda cos site; allows packaging into phage lambda
particles"
terminator complement(11159..11190)
/label=tonB terminator
/note="bidirectional E. coli tonB-P14 transcription
terminator"
protein_bind 11209..11242
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
CDS complement(11352..12008)
/label=HIS3
/note="imidazoleglycerol-phosphate dehydratase, required
for histidine biosynthesis"
promoter complement(12009..12196)
/label=HIS3 promoter
misc_feature complement(12197..12700)
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 12703..12721
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
regulatory 12746..13068
/label=spiralin promoter
/note="spiralin promoter"
/regulatory_class="promoter"
CDS 13070..13666
/label=PuroR
/note="puromycin N-acetyltransferase"
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