Human UNKL ORF clone (NM_001372107) (Cat. No.: V033766)

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Basic Information

Name:: pLV-Tet-miniCMV-mCherry-Puro-UNKL(human)-HA-GFP nanobody

Accession ID:: NM_001372107.1

Antibiotic Resistance:: Ampicillin

Length:: 13356 bp

Type:: Protein expression, Tetracycline inducible

Replication origin:: ori

Host:: Mammalian cells, Lentivirus

Selection Marker:: Puro;mCherry

Promoter:: Tet-miniCMV

5' Primer:: PFLAG-CMV-F

Fusion Tag:: HA-GFP nanobody

Growth Strain(s):: Stbl3

Growth Temperature:: 37℃

Expression Method:: Tetracycline inducible

Gene Synonyms:: C16orf28; ZC3H5L; ZC3HDC5L

Transcript Definition:: Homo sapiens unk like zinc finger (UNKL), transcript variant 11, mRNA

Cat. No.: V033766 pLV-Tet-miniCMV-mCherry-Puro-UNKL(human)-HA-GFP nanobody

$ 299.1

In stock, 1-2 weeks for quality controls

Two tubes of transfection grade plasmid, each tube is about 100 µg (lyophilized).

Transcript ID

NM_001193388.4 NM_001372107.1

Fusion Tag

HA HA-GFP nanobody

Selection Marker

Neo/G418 Puro;mCherry

Expression Method

Transient Tetracycline inducible

Resources

Plasmid Protocol

Plasmid Reconstitution Protocol

Centrifuge at 5,000×g for 5 min.
Carefully open the tube and add sterile water to dissolve the DNA: add 20 μl for 5 μg plasmid, and 100 μl for 100 μg plasmid.
Close the tube and incubate for 10 minutes at room temperature.
Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
Store the plasmid at -20 ℃.
The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

Human UNKL ORF clone (NM_001372107) (Cat. No.: V033766) Annotation

LOCUS       V033766                13356 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V033766
VERSION     V033766
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     misc_feature    25..142
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     protein_bind    210..228
                     /bound_moiety="tetracycline repressor TetR"
                     /gene="tetO"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /label="tet operator"
     protein_bind    245..263
                     /bound_moiety="tetracycline repressor TetR"
                     /gene="tetO"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /label="tet operator"
     protein_bind    281..299
                     /bound_moiety="tetracycline repressor TetR"
                     /gene="tetO"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /label="tet operator"
     protein_bind    317..335
                     /bound_moiety="tetracycline repressor TetR"
                     /gene="tetO"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /label="tet operator"
     protein_bind    352..370
                     /bound_moiety="tetracycline repressor TetR"
                     /gene="tetO"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /label="tet operator"
     protein_bind    388..406
                     /bound_moiety="tetracycline repressor TetR"
                     /gene="tetO"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /label="tet operator"
     promoter        418..456
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="minimal CMV promoter"
     promoter        524..542
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             614..2812
                     /label="UNKL(NM_001372107)"
                     /note="UNKL(NM_001372107)"
                     /gene="UNKL"
     CDS             2813..2839
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2885..3226
                     /codon_start=1
                     /product="GFP-binding fragment of a single-chain camelid
                     antibody (Rothbauer et al., 2008)"
                     /transl_table=1
                     /translation="VQLVESGGALVQPGGSLRLSCAASGFPVNRYSMRWYRQAPGKERE
                     WVAGMSSAGDRSSYEDSVKGRFTISRDDARNTVYLQMNSLKPEDTAVYYCNVNVGFEYW
                     GQGTQVTVSS"
                     /label="GFP nanobody"
     promoter        3352..3851
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             3872..4579
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
                     /label="mCherry"
     CDS             4580..4636
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /transl_table=1
                     /translation="ATNFSLLKQAGDVEENPGP"
                     /label="P2A"
     CDS             4637..5236
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    5247..5835
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5898..6131
                     /note="self-inactivating 3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     promoter        complement(6156..6174)
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /label="SP6 promoter"
     promoter        6963..7067
                     /gene="bla"
                     /label="AmpR promoter"
     CDS             7068..7928
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     rep_origin      8099..8687
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     promoter        8933..9262
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     intron          10396..10461
                     /note="SV40 (simian virus 40) small t antigen intron"
                     /label="small t intron"
     CDS             10591..10611
                     /codon_start=1
                     /locus_tag=""
                     /note=""
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /protein_id=""
                     /transl_table=1
                     /translation="PKKKRKV"
                     /label="SV40 NLS"
     polyA_signal    11036..11170
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     LTR             11339..11972
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    12019..12144
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    12636..12869
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             13054..13098
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"