Step 1: Choose Expression Vector
Select host organism and plasmid backbone
Step 2: Find Your Target Protein
Search databases for the protein you want to express
Comprehensive Database Search
- Enter a gene symbol — Use official symbols like CD8A, EGFR, INS, ACTB
- Watch the trace — See queries to UniProt, NCBI Gene, Ensembl, and PubMed
- Get cDNA info — RefSeq mRNA IDs and Ensembl transcripts are automatically fetched
- Review references — Key literature is attached for traceability
What you get: UniProt ID, NCBI Gene ID, Ensembl ID, RefSeq mRNA accessions, key PubMed references — all traced and linked.
Quick search (with full tracing):
CD8A (T-cell co-receptor)
EGFR (kinase)
Insulin
Beta-actin
Enter a gene symbol or ID and click "Search All Databases"
Step 3: Select Domain & Generate DNA
Choose which part of the protein to express, then get the DNA sequence
What to do here (3 steps):
- Click a domain in the protein map below to select it (e.g., "Ig-like V-type" for CD8)
- Click "Generate DNA" to back-translate the amino acid sequence with E. coli optimized codons
- Review restriction sites — We'll find which enzymes can be used for cloning
Why select a domain? Full-length proteins often don't express well. Domains are functional units that fold independently and are easier to produce.
EGFR - Epidermal growth factor receptor
P00533 | 1210 aa | Homo sapiens
Available Domains
Selected Region
Click a domain to select
DNA Sequence
Select a domain, then click "Generate DNA"
Step 4: What's the Purpose?
Select your expression goal
Express Protein
Recombinant expression for purification or functional studies
Create Fusion
Fuse with tags, reporters, or other proteins
Add Tags
Add purification or detection tags
Mutagenesis
Introduce point mutations or deletions
Truncation
Express specific domain or fragment
Reporter Assay
Fuse with GFP, luciferase, or other reporters
Step 5: Set Cloning Parameters
Configure how the DNA will be assembled
What to do here:
- Choose assembly method — How will you physically join the DNA pieces?
- Blacklist enzymes — Mark sites that must NOT appear in your insert
- Add purification tags — 6xHis is standard for IMAC purification
Recommended: Use Restriction/Ligation with NdeI/XhoI for pET vectors, or Gibson Assembly for seamless cloning.
🔧 Assembly Method
Gibson Assembly
Golden Gate
Restriction/Ligation
SLIC
TOPO Cloning
✂️ Enzyme Blacklist (sites to avoid)
EcoRI
BamHI
HindIII
NdeI
XhoI
NcoI
BsaI
BbsI
🏷️ Tags to Include
6xHis (N-term)
6xHis (C-term)
GST
MBP
SUMO
FLAG
HA
Strep-tag II
⚙️ Additional Options
% min
—
% max
Design Construct
View and edit your plasmid design
Load Example:
pET-28a-EGFR-kinase
Features (0)
Validation Results
Quality checks for your construct design
Export Construct
Download your design in various formats
Preview
Provenance & Sources