Mini-Tn4001PSpuro vector (V009847)

Basic Vector Information

      • Vector Name:
      • Mini-Tn4001PSpuro
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 5321 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Algire MA, Lartigue C, Thomas DW, Assad-Garcia N, Glass JI, Merryman C.

Mini-Tn4001PSpuro vector Vector Map

Mini-Tn4001PSpuro5321 bp6001200180024003000360042004800pactnpT7 promoterM13 fwdf1 oriAmpR promoterblaoriCAP binding sitelac promoterlac operatorM13 revT3 promoter

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

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                5321 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector Mini-Tn4001PSpuro, complete sequence.
ACCESSION   FJ872396
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5321
                     /organism="Cloning vector Mini-Tn4001PSpuro"
                     /mol_type="other DNA"
                     /db_xref="taxon:640477"
     CDS             complement(1..600)
                     /codon_start=1
                     /gene="pac"
                     /product="puromycin N-acetyltransferase"
                     /label=pac
                     /note="puromycin resistance; codon optimized for Mycoplasma
                     genitalium"
                     /protein_id="ACR82772.1"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     gene            complement(1..600)
                     /gene="pac"
                     /label=pac
     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
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2962..3066
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3067..3927
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="ampicillin resistance"
                     /protein_id="ACR82774.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            3067..3927
                     /gene="bla"
                     /label=bla
     rep_origin      4098..4686
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4974..4995
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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
                     /bound_moiety="lac repressor encoded by lacI"
                     /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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