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Table 1 Features of the pIN-G reporter system. Features of the pIN-G Trafficking Reporter Plasmid. Nucleotide positions of the corresponding features of pIN-G are indicated in the right hand column. Unique restriction sites for the extracellular and intracellular Multiple Cloning Sites (MCS-1 and MCS-2, respectively) are italicised. Asterisks denote enzymes yielding blunt ends.

From: PIN-G – A novel reporter for imaging and defining the effects of trafficking signals in membrane proteins

Feature

Position (bp)

Human CMV promoter

1–589

Enhancer region

59–465

AP1 binding site

400–407

CAAT box

521–525

TATA box

554–560

Transcription start site

583

Kozak translation initiation site

606–612

pIN-G start codon

613–615

Igκ-chain leader sequence

613–675

Haemagglutinin epitope tag

676–702

Unique Pvu I site

708

Enhanced Green Fluorescent Protein (EGFP)

742–1458

MCS-1 (BsrGI,AccIII,XhoI,Ecl136I*,SacI,HindIII,EcoRI,PstI,SalI)

1451–1504

c-Myc epitope tag

1504–1533

PDGFR transmembrane domain

1537–1685

MCS-2 (KpnI, SacII,ApaI, SmaI*,BamHI,XbaI,BclI)

1686–1718

pIN-G stop codon

1723–1725

SV40 early mRNA polyadenylation signals

1861–1866,1890–1895

mRNA ends

1899,1911

F1 single-strand DNA origin

1958–2413

Kanr bacterial promoter

2475–2480,2498–2503

Kanr transcription start point

2510

SV40 early promoter (tandem 72 bp)

2587–2730

SV40 origin of replication

2754–2889

Kanamycin/neomycin resistance gene

2938–3732

Herpes simplex thymidine kinase poly-A signal

3968–3973,3981–3986

pUC plasmid origin of replication

4317–4960