T7-TPase vector (V010662)

Basic Vector Information

      • Vector Name:
      • T7-TPase
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 5338 bp
      • Type:
      • Unspecified
      • Copy Number:
      • High Copy
      • Promoter:
      • T7
      • Cloning Method:
      • Restriction Enzyme
      • 5' Primer:
      • M13 Universal
      • 3' Primer:
      • M13 reverse

T7-TPase vector Vector Map

T7-TPase5338 bp6001200180024003000360042004800f1 oriIn lacZ geneT7T7 TermT3M13 revlac operatorlac promoterCAP binding siteL4440oriAmpRAmpR 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 ℃.

T7-TPase vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                5338 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  For in vitro transcription of Tol2 transposase using T7 Promoter.
ACCESSION   .
VERSION     .
KEYWORDS    T7-TPase
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5338)
  AUTHORS   Khattak S, Murawala P, Andreas H, Kappert V, Schuez M, 
            Sandoval-Guzman T, Crawford K, Tanaka EM
  TITLE     Optimized axolotl (Ambystoma mexicanum) husbandry, breeding, 
            metamorphosis, transgenesis and tamoxifen-mediated recombination.
  JOURNAL   Nat Protoc. 2014 Mar;9(3):529-40. doi: 10.1038/nprot.2014.040. Epub 
            2014 Feb 6.
  PUBMED    24504478
REFERENCE   2  (bases 1 to 5338)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nprot.2014.040"; journalName: "Nat Protoc"; date: 
            "2014-03"; volume: "9"; issue: "3"; pages: "529-40"
FEATURES             Location/Qualifiers
     source          1..5338
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     649..668
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        649..667
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3013..3031)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3027..3074
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3127..3147)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3127..3145)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3189..3205)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3189..3205)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3202..3224)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3213..3229
                     /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)."
     promoter        complement(3237..3267)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3282..3303
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3420..3437)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3591..4179)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3671..3690)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4350..5210)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4973..4992
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5211..5315)
                     /gene="bla"
                     /label=AmpR promoter

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