Basic Vector Information
- Vector Name:
- pmycYACTn
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10135 bp
- Type:
- Tri-shuttle cloning vector
- Replication origin:
- ori
- 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
pmycYACTn 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 ℃.
pmycYACTn vector Sequence
LOCUS 40924_32928 10135 bp DNA circular SYN 18-DEC-2018
DEFINITION Tri-shuttle cloning vector pmycYACTn, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10135)
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 10135)
AUTHORS Benders GA, Gibson DG, 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 10135)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10135)
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..10135
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 84..1256
/codon_start=1
/product="IS256 transposase"
/label=IS256 transposase
/protein_id="ADE19325.1"
/translation="MTQVHFTLKSEEIQSIIEYSVKDDVSKNILTTVFNQLMENQRTEY
IQAKEYERTENRQSQRNGYYERSFTTRVGTLELKVPRTRDGHFSPTVFERYQRNEKALM
ASMLEMYVSGVSTRKVSKIVEELCGKSVSKSFVSSLTEQLEPMVNEWQNRLLSEKNYPY
LMTDVLYIKVREENRVLSKSCHIAIGITKDGDREIIGFMIQSGESEETWTTFFEYLKER
GLQGTELVISDAHKGLVSAIRKSFTNVSWQRCQVHFLRNIFTTIPKKNSKSFREAVKGI
FKFTDINLAREAKNRLIHDYIDQPKYSKACASLDDGFEDAFQYTVQGNSHNRLKSTNLI
ERLNQEVRRREKIIRIFPNQTSANRLIGAVLMDLHDEWIYSSRKYINFDK"
promoter 1256..1326
/label=AmpR promoter
CDS 1327..2184
/label=AmpR
/note="beta-lactamase"
rep_origin 2358..2946
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
mobile_element 3103..3128
/mobile_element_type="insertion sequence:IS256"
/label=insertion sequence:IS256
/note="outer inverted repeat"
CDS complement(3138..6182)
/label=lacZ
/note="beta-galactosidase"
regulatory complement(6219..6463)
/label=spiralin promoter
/note="spiralin promoter"
/regulatory_class="promoter"
gene complement(6464..8428)
/gene="tetM"
/label=tetM
CDS complement(6494..8428)
/codon_start=1
/gene="tetM"
/product="tetM"
/label=tetM
/protein_id="ADE19326.1"
/translation="MEENHMKIINIGVLAHVDAGKTTLTESLLYNSGAITELGSVDKGT
TRTDNTLLERQRGITIQTGITSFQWENTKVNIIDTPGHMDFLAEVYRSLSVLDGAILLI
SAKDGVQAQTRILFHALRKMGIPTIFFINKIDQNGIDLSTVYQDIKEKLSAEIVIKQKV
ELYPNVCVTNFTESEQWDTVIEGNDDLLEKYMSGKSLEALELEQEESIRFQNCSLFPLY
HGSAKSNIGIDNLIEVITNKFYSSTHRGPSELCGNVFKIEYTKKRQRLAYIRLYSGVLH
LRDSVRVSEKEKIKVTEMYTSINGELCKIDRAYSGEIVILQNEFLKLNSVLGDTKLLPQ
RKKIENPHPLLQTTVEPSKPEQREMLLDALLEISDSDPLLRYYVDSTTHEIILSFLGKV
QMEVISALLQEKYHVEIEITEPTVIYMERPLKNAEYTIHIEVPPNPFWASIGLSVSPLP
LGSGMQYESSVSLGYLNQSFQNAVMEGIRYGCEQGLYGWNVTDCKICFKYGLYYSPVST
PADFRMLAPIVLEQVLKKAGTELLEPYLSFKIYAPQEYLSRAYNDAPKYCANIVDTQLK
NNEVILSGEIPARCIQEYRSDLTFFTNGRSVCLTELKGYHVTTGEPVCQPRRPNSRIDK
VRYMFNKIT"
regulatory complement(8435..8757)
/label=spiralin promoter
/note="spiralin promoter"
/regulatory_class="promoter"
misc_feature 8758..9261
/label=CEN/ARS
/note="S. cerevisiae CEN6 centromere fused to an
autonomously replicating sequence"
promoter 9262..9449
/label=HIS3 promoter
CDS 9450..10106
/label=HIS3
/note="imidazoleglycerol-phosphate dehydratase, required
for histidine biosynthesis"
mobile_element complement(10110..10135)
/mobile_element_type="insertion sequence:IS256"
/label=insertion sequence:IS256
/note="inner inverted repeat"
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