pT3-myr-AKT-HA vector (V005932)

Basic Vector Information

      • Vector Name:
      • pT3-myr-AKT-HA
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 7091 bp
      • Type:
      • Mammalian Expression
      • Cloning Method:
      • Gateway Cloning
      • 5' Primer:
      • EF-1a-Fwd
      • 3' Primer:
      • pT3EF1a-R 5'-TGACACCTACTCAGACAATG

pT3-myr-AKT-HA vector Vector Map

pT3-myr-AKT-HA7091 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900EF-1-alpha promoterT7 promoterattB1myrAkt1HAattB2V5 tagbGH poly(A) signalloxPpBABE 3'M13 fwdpRS-markerpGEX 3'pBRforEcoAmpR promoterAmpRoriL4440CAP binding sitelac promoterlac operatorM13 revEBV-revloxP

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 ℃.

pT3-myr-AKT-HA vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_42264        7091 bp DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7091)
  AUTHORS   Calvisi DF, Wang C, Ho C, Ladu S, Lee SA, Mattu S, Destefanis G, 
            Delogu S, Zimmermann A, Ericsson J, Brozzetti S, Staniscia T, Chen 
            X, Dombrowski F, Evert M
  TITLE     Increased lipogenesis, induced by AKT-mTORC1-RPS6 signaling, 
            promotes development of human hepatocellular carcinoma.
  JOURNAL   Gastroenterology. 2011 Mar;140(3):1071-83. Epub 2010 Dec 11.
  PUBMED    21147110
REFERENCE   2  (bases 1 to 7091)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7091)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Gastroenterology."; date: "2011-03"; volume: "140(3)"; pages: 
            "1071-83. Epub 2010 Dec 11"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7091
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..1179
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     promoter        1196..1214
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1293..1317
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1338..1358
                     /codon_start=1
                     /label=myr
                     /note="N-myristoylation signal from Src kinase (Pellman et
                     al., 1985; Kaplan et al., 1988)"
                     /translation="MGSSKSK"
     CDS             1371..2810
                     /codon_start=1
                     /label=Akt1
                     /note="human serine/threonine protein kinase activated by
                     growth factors"
                     /translation="MNDVAIVKEGWLHKRGEYIKTWRPRYFLLKNDGTFIGYKERPQDV
                     DQRESPLNNFSVAQCQLMKTERPRPNTFIIRCLQWTTVIERTFHVETPEEREEWATAIQ
                     TVADGLKRQEEETMDFRSGSPSDNSGAEEMEVSLAKPKHRVTMNEFEYLKLLGKGTFGK
                     VILVKEKATGRYYAMKILKKEVIVAKDEVAHTLTENRVLQNSRHPFLTALKYSFQTHDR
                     LCFVMEYANGGELFFHLSRERVFSEDRARFYGAEIVSALDYLHSEKNVVYRDLKLENLM
                     LDKDGHIKITDFGLCKEGIKDGATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMY
                     EMMCGRLPFYNQDHEKLFELILMEEIRFPRTLGPEAKSLLSGLLKKDPTQRLGGGSEDA
                     KEIMQHRFFANIVWQDVYEKKLSPPFKPQVTSETDTRYFDEEFTAQMITITPPDQDDSM
                     ECVDSERRPHFPQFSYSASGTA"
     CDS             2811..2837
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     protein_bind    complement(2857..2881)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2934..2975
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    3019..3243
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    3313..3346
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     3402..3422
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(3804..3820)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4029..4048)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     4148..4170
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4208..4226)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     promoter        4294..4398
                     /label=AmpR promoter
     CDS             4399..5256
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5430..6018
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6172..6189
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6306..6327
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6342..6372
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6380..6396
                     /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     6404..6420
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6846..6865)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(6918..6951)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        7091
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"

This page is informational only.