Last data update: 24 January 2024 16:39 CET
Plasmid name: pUChGNTI2 (LMBP 3173)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner core plasmid |
| GeneCorner sequence: |
p3173.gb
(View with Genome Compiler) p3173.pdf |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human N-acetylglucosaminyltransferase I cDNA (GlcNAc-TI, MGAT1) Streptavidin-affinity tag no. 111 (strep-tag); C-terminal |
| Promoter: | Escherichia coli lac operon promoter |
| Ribosome binding site: |
Ribosome binding site (RBS) of the Escherichia coli lac Z gene (lacZ) |
| Terminator: | - |
| Selection marker: | Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin |
| Host range: | Escherichia coli |
| Parental clone: | pUChGNTI2 |
| Further information: | This plasmid was derived from pUChGNTI1 by site-specific mutagenesis according to Morinaga et al. (1984), inserting the steptavidin-affinity tag no. 111 (strep-tag) at the C-terminus of the human N-acetylglucosaminyltransferase I cDNA (hGlcNAc-TI). The inserted strep-tag has been synthesized according to Schmidt et al. (1993) but with a different termination codon (TGA instead of TAA), making the short peptide tag removable as a BclI-Eco47III fragment. The Strep-tag protein has been succesfully used for the detection of the recombinant human N-acetylglucosaminyltransferase I expressed in Escherichia coli. When cloning a fragment downstream from the lac promoter it may be advisable to use lacI(q) strains in order to prevent fortuitous expression of a possibly noxious polypeptide. The nucleotide sequence of the human GlcNAc-TI cDNA corresponds with the Genbank accession number M55621.1. |
| EMBL Accession number: | M55621.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 16/12/1994 |
| Sequence detail: | Nucleotide sequence at the start of the human GlcNAc-TI coding sequence:
pUC18 --> --> PCR fragment
5' TCTAGAGTC T CCAGG ATG.CTG.AAG. 3'
XbaI --- ^ *
blunt-ended HindII
^: T-residue, due to tailing.
*: Start codon of the human GlcNAc-TI coding sequence.
Nucleotide sequence at the end of the human GlcNAc-TI coding sequence:
insert
-----------------------------------------------
Trp Asn Ser Ala Trp Arg His Pro Gln Phe Gly Gly
5' TGG.AAT.AGC.GCT.TGG.CGT.CAC.CCG.CAG.TTC.GGT.GGT.TGA TCA TAG 3'
Eco47III BclI
* ^
-----------------------------------
strep-tag sequence
*: Termination codon inserted via site-specific mutagenesis.
^: Termination codon. |
| Authenticity test: | Restriction enzyme pattern analysed at GeneCorner: BglI, Eco47III, HindII and NarI/XmnI. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Dr M. Maras(1)(2) and Prof. Dr R. Contreras(1)(2). (1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | - |
| Related plasmid reference: | Schmidt et al., Protein Eng. 6 (1993), 109-122 [PMID: 8433964] Morinaga et al., Biotechnology 2 (1984), 636-639 Kumar et al., Proc. Natl. Acad. Sci. U.S.A. 87 (1990), 9948-9952 [PMID: 1702225] |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 MC1061 |
| Host reference: | Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493] |
| Related host reference: | Brigé et al., Biochem. J. 394 (2006), 335-344 [PMID: 16293111] |
| Cultivation medium: | LB-Lennox + ampicillin (100 μg/ml) |
| Cultivation temperature: | 37°C |
| Biosafety level: | L1 |
| Cultivation remark: | - |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pUChGNTI2 (LMBP 3173) is available at BCCM/GeneCorner. This plasmid was deposited by Dr M. Maras and Prof. Dr R. Contreras. |
Note: Up-to-date, validated data are enclosed with the biological material. Nevertheless, these data are a snapshot at a given moment; further updates are always possible.