pTag 2.0 vector (V002816)

Basic Vector Information

      • Vector Name:
      • pTag 2.0
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 6284 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Vidigal J, Fernandes B, Dias MM, Patrone M, Roldao A, Carrondo MJT., Alves PM, Teixeira AP.

pTag 2.0 vector Vector Map

pTag 2.06284 bp30060090012001500180021002400270030003300360039004200450048005100540057006000M13 fwdOpIE-2 promoterFRTEGFPbeta-globin poly(A) signalSV40 poly(A) signalHygROpIE-1 promoterM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR 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 ℃.

pTag 2.0 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_42534        6284 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pTag 2.0, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6284)
  AUTHORS   Vidigal J, Fernandes B, Dias MM, Patrone M, Roldao A, Carrondo MJT.,
            Alves PM, Teixeira AP.
  TITLE     RMCE-based insect cell platform to produce membrane proteins 
            captured on HIV-1 Gag virus-like particles
  JOURNAL   Appl. Microbiol. Biotechnol. (2017) In press
  PUBMED    29143881
REFERENCE   2  (bases 1 to 6284)
  AUTHORS   Vidigal J.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-OCT-2017) Animal Cell Technology Unit, Ibet, Av. da 
            Republica, ITQB building, Oeiras 2780-157, Portugal
REFERENCE   3  (bases 1 to 6284)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6284)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Microbiol. Biotechnol. (2017) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (30-OCT-2017) Animal Cell Technology Unit, Ibet, Av. da Republica, 
            ITQB building, Oeiras 2780-157, Portugal"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        443..990
                     /label=OpIE-2 promoter
                     /note="strong constitutive baculovirus promoter for insect
                     cell expression"
     protein_bind    1027..1074
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             1101..1817
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    1972..2027
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     polyA_signal    2442..2563
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(2591..3613)
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(3704..3995)
                     /label=OpIE-1 promoter
                     /note="moderate constitutive baculovirus promoter for
                     insect cell expression"
     primer_bind     complement(4063..4079)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4087..4103)
                     /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(4111..4141)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4156..4177)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4465..5053)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5227..6084)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6085..6189)
                     /label=AmpR promoter

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