HA-p62 vector (V007450)

Basic Vector Information

      • Vector Name:
      • HA-p62
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 6414 bp
      • Type:
      • Mammalian Expression
      • Selection Marker:
      • Zeocin
      • Copy Number:
      • High Copy
      • Cloning Method:
      • Restriction Enzyme
      • 5' Primer:
      • CMV-F
      • 3' Primer:
      • BGH-rev

HA-p62 vector Vector Map

HA-p626414 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300CAP binding sitelac promoterlac operatorM13 revSV40 poly(A) signalBleoREM7 promoterSV40 promoterf1 oribGH poly(A) signalHALNCXtet operatortet operatorCMV promoterCMV enhancerpRS-markerAmpR promoterAmpRoriL4440

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 ℃.

HA-p62 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                6414 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression of HA tagged human p62.
ACCESSION   .
VERSION     .
KEYWORDS    HA-p62
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6414)
  AUTHORS   Fan W, Tang Z, Chen D, Moughon D, Ding X, Chen S, Zhu M, Zhong Q
  TITLE     Keap1 facilitates p62-mediated ubiquitin aggregate clearance via 
            autophagy.
  JOURNAL   Autophagy. 2010 Jul 22. 6(5):.
  PUBMED    20495340
REFERENCE   2  (bases 1 to 6414)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Autophagy. 
            2010 Jul 22. 6(5):."
FEATURES             Location/Qualifiers
     source          1..6414
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    270..391
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(281..300)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     335..354
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(521..895)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(914..961)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(1009..1338)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1023..1158
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1077..1096)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1323..1343
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1352..1780)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1462..1483)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1674..1693
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    1826..2050
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2039..2056
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2097..2123)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3585..3609)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    3619..3637
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3640..3658
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(3660..3863)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3689..3709)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3864..4243
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4415..4434
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4509..4613
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4614..5474
                     /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     complement(4832..4851)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5645..6233
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6134..6153
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6387..6404
                     /label=L4440
                     /note="L4440 vector, forward primer"

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