Last data update: 24 January 2024 16:39 CET
Plasmid name: pTM2HIFNG (LMBP 1621)
| New search | Print data sheet |
| Price category: | Cat. 1 (cf. price list) |
| Status: | GeneCorner non-core plasmid |
| Depositor's sequence: | p1621.gb |
| Sequence analysis results Genecorner: |
- |
| Cloned DNA: |
Human interferon γ cDNA (IFNG); truncated and mutated mature sequence |
| Promoter: | Phage λ major leftward promoter (λ PL) |
| Ribosome binding site: |
Ribosome binding site (RBS) of the Escherichia coli tryptophan operon attenuator peptide (trp) |
| Terminator: | Phage fd terminator |
| Selection marker: | Ampicillin (amp) |
| Replicon: | Escherichia coli plasmid pMB1 origin Phage f1 origin |
| Host range: | Escherichia coli; use strains with a cI function |
| Parental clone: | pPLc321HIFNGv; pMa58 |
| Further information: | The plasmid was constructed by inserting the SalI/XmnI fragment of pMa58 into the unique SalI and unique XmnI sites of pPLc321HIFNGv. pTM2HIFNG is a Target Mutagenesis plasmid. The human interferon γ gene (IFNG) is a truncated and mutated variant, derived from the ancestral pPLc321HIFNGHS vector; it contains an extra HhaI site by silent mutations of codons 130 and 131, and a unique SalI site at codons 138-140. The SalI site is used for ligation to pMa58, resulting in the loss of 4 carboxy terminal codons of human IFNG. After induction at 42°C, this plasmid gives rise to a larger human IFNG peptide, due to the read-through (at the SalI-junction). The plasmid should not be used for expression as the human IFNG protein has additional nonsense information at its carboxy terminal end. This plasmid can be used in conjunction with pTM1HIFNG to obtain selection for the opposite strand. The nucleotide sequence of the wt human IFNG DNA corresponds with the EMBL Sequence Database accession number AM903379.1. Other names of the plasmid are pTM2G and pMutHIIFam-. |
| EMBL Accession number: | AM903379.1, view at EMBL, GenBank, DDBJ |
| Latest sequence update: | 19/06/1989 |
| Sequence detail: | Start of the mature human IFN gamma cDNA:
|-> mature human IFNG
| 1 5
5' AAAAAGGGTATCGATTCC.ATG.CAG.GAC.CCA.TAT.GTA.AAA 3'
ClaI Met Gln Asp Pro Tyr Val Lys
Nucleotide sequence at the end of the human IFNG wt DNA:
130 139 140 143
5' CGA.AAA.AGG.AGT.CAG.ATG.CTG.TTT.CGA.GGT.CGA.AGA.GCA.TCC.CAG.TAA.TGGTTGTC 3'
Arg Lys Arg Ser Gln Met Leu Phe Arg Gly Arg Arg Ala Ser Gln
Nucleotide sequence at the end of the human IFNG variant:
human IFNG ->|-> pMa58
130 131 139|
5' CGA.AAG.CGC.AGT.CAG.ATG.CTG.TTT.CGA.GGT.CGA.CCT.GCA.GCC.AAG.CTT.GGT.CTA.GAG... 3'
Arg Lys Arg Ser Gln Met Leu Phe Arg Gly Arg Pro Ala Ala Lys Leu Gly Leu Glu
* * * SalI PstI XbaI
HhaI
*: mutated nucleotide |
| Authenticity test: | The plasmid still needs to be subjected to the authenticity test. |
| Class: | Recombinant plasmid |
| Type: | Plasmid |
| History of deposit: | This plasmid was deposited by Prof. Dr E. Remaut(1) (2). (1) Department for Molecular Biomedical Research, VIB, Ghent, Belgium (2) Department of Biomedical Molecular Biology, Ghent University, Ghent, Belgium |
| Plasmid reference: | PhD thesis Erwin Sablon (1990) |
| Restricted distribution: | - BCCM MTA |
| Distributed as: | H/P active culture or plasmid DNA |
| Host for distribution: | Escherichia coli K12 MC1061(λ) |
| Host reference: | Mertens et al., Gene 164 (1995), 9-15 [PMID: 7590329] |
| Related host reference: | Casadaban et al., J. Mol. Biol. 138 (1980), 179-207 [PMID: 6997493] |
| Cultivation medium: | LB-Lennox + ampicillin (100 μg/ml) |
| Cultivation temperature: | 28°C |
| Biosafety level: | L1 |
| Other culture collection numbers: | - |
Refer in your Materials and Methods: |
pTM2HIFNG (LMBP 1621) is available at BCCM/GeneCorner. This plasmid was deposited by Prof. Dr E. Remaut and was published in Erwin Sablon, 1990. |
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.