Human KREMEN1 Full-length Gene in Lentivector, Endotoxin-free DNA

Description

KREMEN (kringle containing protein marking the eye and the nose) proteins, including KREMEN1 and KREMEN2, are single pass type I transmembrane proteins that contain extracellular kringle, WSC, and CUB domains. KREMEN proteins cooperate with dickkopf  (Dkk) proteins to antagonize wingless (WNT)/beta-catenin signaling via formation of a KREMEN-Dkk-LRP (LDL-receptor-related protein ternary complex) that triggers the internalization and clearance of the complex from the cell surface. KREMEN1 is a high-affinity Dkk-1 receptor and a component of a ternary membrane complex consisting of Wnt, the 7-transmembrane domain receptor Frizzled, and LRP5/6 co-receptor to block canonical WNT signaling. Dkk-1 and Dkk-4 bind directly to the LRP5/6 co-receptor to antagonize the canonical Wnt signaling pathway, but not the planar cell polarity (PCP) signaling pathway that does not require LRP5/6. In contrast, Dkk-3 has no effect on Wnt signaling and Dkk-2 can function either as an LRP agonist or antagonist, depending on whether the cell expresses Kremen proteins. All three extracellular domains of KREMEN1 are required for binding to the cysteine-rich domain of Dkk-1.

 

 

Gene Symbol: KREMEN1; KRM1; KREMEN; Kremen-1

 

NCBI Gene ID: 83999

 

Uniprot Entry: Q96MU8

 

Construct Details: Full length human KREMEN1 gene (encoding Uniprot Accession #Q96MU8-3) is subcloned into the Lentiviral expression vector pLTC with an upstream CMV promoter and with or without a selection marker, which can be used for both transient and stable expression in mammalian cells.  It can be co-transfected with the LentiPAK DNA mix (SKU# LP-001) into HEK293 cells to produce high titer lentiviral particles.  Multiple choices of a stable antibiotic selection marker are available.

 

Vector Type: pLTC or pLTC-IRES-Marker (lentiviral expression vector containing a heterologous CMV promoter, see the vector map above)

 

Formulation: Supplied with 10 μg of endotoxin-free plasmid DNA at 200 ng/μl in sterile TE buffer (pH8.0)

 

Gene Insert Size: 1377 (bp)

 

Gene Insert Sequence:  

ATGGCGCCGCCCGCCGCCCGCCTCGCCCTGCTCTCCGCCGCGGCGCTCACGCTGGCGGCCCGGCCCGCGCCTAGCCCCGG

CCTCGGCCCCGGACCCGAGTGTTTCACAGCCAATGGTGCGGATTATAGGGGAACACAGAACTGGACAGCACTACAAGGCG

GGAAGCCATGTCTGTTTTGGAACGAGACTTTCCAGCATCCATACAACACTCTGAAATACCCCAACGGGGAGGGGGGCCTG

GGTGAGCACAACTATTGCAGAAATCCAGATGGAGACGTGAGCCCCTGGTGCTATGTGGCAGAGCACGAGGATGGTGTCTA

CTGGAAGTACTGTGAGATACCTGCTTGCCAGATGCCTGGAAACCTTGGCTGCTACAAGGATCATGGAAACCCACCTCCTC

TAACTGGCACCAGTAAAACGTCCAACAAACTCACCATACAAACTTGCATCAGTTTTTGTCGGAGTCAGAGGTTCAAGTTT

GCTGGGATGGAGTCAGGCTATGCTTGCTTCTGTGGAAACAATCCTGATTACTGGAAGTACGGGGAGGCAGCCAGTACCGA

ATGCAACAGCGTCTGCTTCGGGGATCACACCCAACCCTGTGGTGGCGATGGCAGGATCATCCTCTTTGATACTCTCGTGG

GCGCCTGCGGTGGGAACTACTCAGCCATGTCTTCTGTGGTCTATTCCCCTGACTTCCCCGACACCTATGCCACGGGGAGG

GTCTGCTACTGGACCATCCGGGTTCCGGGGGCCTCCCACATCCACTTCAGCTTCCCCCTATTTGACATCAGGGACTCGGC

GGACATGGTGGAGCTTCTGGATGGCTACACCCACCGTGTCCTAGCCCGCTTCCACGGGAGGAGCCGCCCACCTCTGTCCT

TCAACGTCTCTCTGGACTTCGTCATCTTGTATTTCTTCTCTGATCGCATCAATCAGGCCCAGGGATTTGCTGTTTTATAC

CAAGCCGTCAAGGAAGAACTGCCACAGGAGAGGCCCGCTGTCAACCAGACGGTGGCCGAGGTGATCACGGAGCAGGCCAA

CCTCAGTGTCAGCGCTGCCCGGTCCTCCAAAGTCCTCTATGTCATCACCACCAGCCCCAGCCACCCACCTCAGACTGTCC

CAGGATGGACAGTCTATGGTCTGGCAACTCTCCTCATCCTCACAGTCACAGCCATTGTAGCAAAGATACTTCTGCACGTC

ACATTCAAATCCCATCGTGTTCCTGCTTCAGGGGACCTTAGGGATTGTCATCAACCAGGGACTTCGGGGGAAATCTGGAG

CATTTTTTACAAGCCTTCCACTTCAATTTCCATCTTTAAGAAGAAACTCAAGGGTCAGAGTCAACAAGATGACCGCAATC

CCCTTGTGAGTGACTAA

 

 

Primers: Gene insert coding sequence can be confirmed by following primers:

(a) forward (or 5'-end) primer: 5’-CAAATGGGCGGTAGGCGTG-3’;

(b) reverse (or 3'-end) primer: 5’-CCAGGTTTCCGGGCCCTCAC-3’

 

 

Note: The sequence may vary due to the naturally occurring variants or mutations, such as single nucleotide polymorphism, or the artifact during PCR amplification.  High fidelity PCR systems should always be used. 

 

 

 

 

 

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

Restriction: This product is not transferable or re-sellable.  Customer obtain no right to transfer, assign, or sublicense its use rights, or to transfer, resell, package, or otherwise distribute the product, or to use the product for the benefit of any third party or for any commercial purpose.  Customer may only use the product in compliance with applicable local, state and federal laws, regulations and rules.  Customer may not directly or indirectly use the product or allow the transfer, transmission, export or re-export of all or any part of the product in violation of any export control law or regulation of the united states or any other relevant jurisdiction.  Your use of this product constitutes acceptance of the terms of this limited use agreement.  Please refer to our “terms & conditions” for details.  If you are not willing to accept the limitation of this agreement, G&P Biosciences will accept return of the product for a full/partial refund.

 

 

The product is shipped at 4°C. Upon receipt, centrifuge the product briefly before opening the vial. It is recommended to store small aliquots at the temperature below –20°C for long-term storage and the product is stable for 6 months. 

 

 

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

 

 

 

 

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

 

 

1. Biochim Biophys Acta 1518:63 (2001)

2. Development 129:5587 (2002)

3. Nature 417:664 (2002)

4. J Biol Chem. 283:23371 (2008)

5. Mol Cancer. 9:247 (2010)

 

 

 

 

Product datasheet (pdf) can be downloaded here: LTP1381-PDS.pdf

 

 

 

Additional supporting documents, including COA and MSDS are available upon request.

 

 

 

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

Restriction: This product is not transferable or re-sellable.  Customer obtain no right to transfer, assign, or sublicense its use rights, or to transfer, resell, package, or otherwise distribute the product, or to use the product for the benefit of any third party or for any commercial purpose.  Customer may only use the product in compliance with applicable local, state and federal laws, regulations and rules.  Customer may not directly or indirectly use the product or allow the transfer, transmission, export or re-export of all or any part of the product in violation of any export control law or regulation of the united states or any other relevant jurisdiction.  Your use of this product constitutes acceptance of the terms of this limited use agreement.  Please refer to our “terms & conditions” for details.  If you are not willing to accept the limitation of this agreement, G&P Biosciences will accept return of the product for a full/partial refund.