Basic Vector Information
Mini-Tn4001PSpuro 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 ℃.
Mini-Tn4001PSpuro vector Sequence
LOCUS 40924_1984 5321 bp DNA circular SYN 17-DEC-2018 DEFINITION Cloning vector Mini-Tn4001PSpuro, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5321) AUTHORS Algire MA, Lartigue C, Thomas DW, Assad-Garcia N, Glass JI, Merryman C. TITLE New selectable marker for manipulating the simple genomes of Mycoplasma species JOURNAL Antimicrob. Agents Chemother. 53 (10), 4429-4432 (2009) PUBMED 19687239 REFERENCE 2 (bases 1 to 5321) AUTHORS Merryman C. TITLE Direct Submission JOURNAL Submitted (30-MAR-2009) Synthetic Biology, J. Craig Venter Institute, 9704 Medical Center Dr, Rockville, MD 20850, USA REFERENCE 3 (bases 1 to 5321) TITLE Direct Submission REFERENCE 4 (bases 1 to 5321) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Antimicrob. Agents Chemother."; date: "2009"; volume: "53"; issue: "10"; pages: "4429-4432" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (30-MAR-2009) Synthetic Biology, J. Craig Venter Institute, 9704 Medical Center Dr, Rockville, MD 20850, USA" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..5321 /mol_type="other DNA" /organism="synthetic DNA construct" CDS complement(4..600) /codon_start=1 /label=PuroR /note="puromycin N-acetyltransferase" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" CDS 1105..2277 /codon_start=1 /gene="tnp" /product="transposase" /label=tnp /protein_id="ACR82773.1" /translation="MTQVHFTLKSEEIQSIIEYSVKDDVSKNILTTVFNQLMENQRTEY IQAKEYERTENRQSQRNGYYERSFTTRVGTLELKVPRTRDGHFSPTVFERYQRNEKALM ASMLEMYVSGVSTRKVSKIVEELCGKSVSKSFVSSLTEQLEPMVNEWQNRLLSEKNYPY LMTDVLYIKVREENRVLSKSCHIAIGITKDGDREIIGFMIQSGESEETWTTFFEYLKER GLQGTELVISDAHKGLVSAIRKSFTNVSWQRCQVHFLRNIFTTIPKKNSKSFREAVKGI FKFTDINLAREAKNRLIHDYIDQPKYSKACASLDDGFEDAFQYTVQGNSHNRLKSTNLI ERLNQEVRRREKIIRIFPNQTSANRLIGAVLMDLHDEWIYSSRKYINFDK" gene 1105..2277 /gene="tnp" /label=tnp promoter complement(2298..2316) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2323..2339) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin 2481..2936 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 2962..3066 /label=AmpR promoter CDS 3067..3924 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 4098..4686 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 4974..4995 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 5010..5040 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5048..5064 /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)." primer_bind 5072..5088 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" promoter 5109..5127 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase"
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