Basic Vector Information
- Vector Name:
- miniTn4001-Puro-1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5311 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Paetzold B, Carolis C, Ferrar T, Serrano L, Lluch-Senar M.
miniTn4001-Puro-1 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 ℃.
miniTn4001-Puro-1 vector Sequence
LOCUS 40924_1994 5311 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector miniTn4001-Puro-1, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5311)
AUTHORS Paetzold B, Carolis C, Ferrar T, Serrano L, Lluch-Senar M.
TITLE In situ overlap and sequence synthesis during DNA assembly
JOURNAL ACS Synth Biol 2 (12), 750-755 (2013)
PUBMED 24161008
REFERENCE 2 (bases 1 to 5311)
AUTHORS Paetzold B.
TITLE Direct Submission
JOURNAL Submitted (21-MAR-2013) EMBL-CRG Systems Biology Research Unit,
Centre for Genomic Regulation, Dr. Aiguader 88, Barcelona 08003,
Spain
REFERENCE 3 (bases 1 to 5311)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5311)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
Biol"; date: "2013"; volume: "2"; issue: "12"; pages: "750-755"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(21-MAR-2013) EMBL-CRG Systems Biology Research Unit, Centre for
Genomic Regulation, Dr. Aiguader 88, Barcelona 08003, Spain"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..5311
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(2..18)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
mobile_element complement(27..52)
/mobile_element_type="insertion sequence:IS256"
/label=insertion sequence:IS256
promoter complement(74..92)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(113..129)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(137..153)
/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(161..191)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(206..227)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(515..1103)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1277..2134)
/label=AmpR
/note="beta-lactamase"
promoter complement(2135..2239)
/label=AmpR promoter
rep_origin complement(2265..2720)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 2862..2878
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 2888..2906
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 3008..4180
/codon_start=1
/product="transposase"
/label=transposase
/protein_id="AHL28656.1"
/translation="MTQVHFTLKSEEIQSIIEYSVKDDVSKNILTTVFNQLMENQRTEY
IQAKEYERTENRQSQRNGYYERSFTTRVGTLELKVPRTRDGHFSPTVFERYQRNEKALM
ASMLEMYVSGVSTRKVSKIVEELCGKSVSKSFVSSLTEQLEPMVNEWQNRLLSEKNYPY
LMTDVLYIKVREENRVLSKSCHIAIGITKDGDREIIGFMIQSGESEETWTTFFEYLKER
GLQGTELVISDAHKGLVSAIRKSFTNVSWQRCQVHFLRNIFTTIPKKNSKSFREAVKGI
FKFTDINLAREAKNRLIHDYIDQPKYSKACASLDDGFEDAFQYTVQGNSHNRLKSTNLI
ERLNQEVRRREKIIRIFPNQTSANRLIGAVLMDLHDEWIYSSRKYINFDK"
mobile_element 4185..4210
/mobile_element_type="insertion sequence:IS256"
/label=insertion sequence:IS256
primer_bind 4222..4238
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
regulatory 4287..4296
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 4627..5223
/label=PuroR
/note="puromycin N-acetyltransferase"
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