OneClick

A Program for Designing Focused Mutagenesis Experiments

OneClick Program

OneClick is a program for designing focused mutagenesis (e.g. site-directed and saturation mutagenesis). The program provides

  1. Sequences of mutagenic primers
  2. PCR sample preparation and thermal cycling conditions
  3. Statistical analysis of protein mutant library

To begin, either upload the DNA sequence file (in FASTA format) of the protein of interest, or directly enter the DNA sequence.

File Upload

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Direct Input

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DNA Sequence (File Upload)

This function allows the users to upload a text file containing a DNA sequence in FASTA format that encodes the gene of interest. A sequence in FASTA format begins with a single-line description, followed by line(s) of sequence data. The description line (defline) is distinguished from the sequence data by a greater-than (">") symbol at the beginning.

An example of FASTA format sequence is provided below:

>Gene encoding for cytochrome P450 BM-3
ATGACAATTAAAGAAATGCCTCAGCCAAAAACGTTTGGAGAGCTTAAAAATTTACCGTTATTAAACACAG
ATAAACCGGTTCAAGCTTTGATGAAAATTGCGGATGAATTAGGAGAAATCTTTAAATTCGAGGCGCCTGG
TCGTGTAACGCGCTACTTATCAAGTCAGCGTCTAATTAAAGAAGCATGCGATGAATCACGCTTTGATAAA
AACTTAAGTCAAGCGCTTAAATTTGTACGTGATTTTGCAGGAGACGGGTTATTTACAAGCTGGACGCATG
AAAAAAATTGGAAAAAAGCGCATAATATCTTACTTCCAAGCTTCAGTCAGCAGGCAATGAAAGGCTATCA
TGCGATGATGGTCGATATCGCCGTGCAGCTTGTTCAAAAGTGGGAGCGTCTAAATGCAGATGAGCATATT
GAAGTACCGGAAGACATGACACGTTTAACGCTTGATACAATTGGTCTTTGCGGCTTTAACTATCGCTTTA
ACAGCTTTTACCGAGATCAGCCTCATCCATTTATTACAAGTATGGTCCGTGCACTGGATGAAGCAATGAA
CAAGCTGCAGCGAGCAAATCCAGACGACCCAGCTTATGATGAAAACAAGCGCCAGTTTCAAGAAGATATC
AAGGTGATGAACGACCTAGTAGATAAAATTATTGCAGATCGCAAAGCAAGCGGTGAACAAAGCGATGATT
TATTAACGCATATGCTAAACGGAAAAGATCCAGAAACGGGTGAGCCGCTTGATGACGAGAACATTCGCTA
TCAAATTATTACATTCTTAATTGCGGGACACGAAACAACAAGTGGTCTTTTATCATTTGCGCTGTATTTC
TTAGTGAAAAATCCACATGTATTACAAAAAGCAGCAGAAGAAGCAGCACGAGTTCTAGTAGATCCTGTTC
CAAGCTACAAACAAGTCAAACAGCTTAAATATGTCGGCATGGTCTTAAACGAAGCGCTGCGCTTATGGCC
AACTGCTCCTGCGTTTTCCCTATATGCAAAAGAAGATACGGTGCTTGGAGGAGAATATCCTTTAGAAAAA
GGCGACGAACTAATGGTTCTGATTCCTCAGCTTCACCGTGATAAAACAATTTGGGGAGACGATGTGGAAG
AGTTCCGTCCAGAGCGTTTTGAAAATCCAAGTGCGATTCCGCAGCATGCGTTTAAACCGTTTGGAAACGG
TCAGCGTGCGTGTATCGGTCAGCAGTTCGCTCTTCATGAAGCAACGCTGGTACTTGGTATGATGCTAAAA
CACTTTGACTTTGAAGATCATACAAACTACGAGCTGGATATTAAAGAAACTTTAACGTTAAAACCTGAAG
GCTTTGTGGTAAAAGCAAAATCGAAAAAAATTCCGCTTGGCGGTATTCCTTCACCTAGCACTGAACAGTC
TGCTAAAAAAGTACGCAAAAAGGCAGAAAACGCTCATAATACGCCGCTGCTTGTGCTATACGGTTCAAAT
ATGGGAACAGCTGAAGGAACGGCGCGTGATTTAGCAGATATTGCAATGAGCAAAGGATTTGCACCGCAGG
TCGCAACGCTTGATTCACACGCCGGAAATCTTCCGCGCGAAGGAGCTGTATTAATTGTAACGGCGTCTTA
TAACGGTCATCCGCCTGATAACGCAAAGCAATTTGTCGACTGGTTAGACCAAGCGTCTGCTGATGAAGTA
AAAGGCGTTCGCTACTCCGTATTTGGATGCGGCGATAAAAACTGGGCTACTACGTATCAAAAAGTGCCTG
CTTTTATCGATGAAACGCTTGCCGCTAAAGGGGCAGAAAACATCGCTGACCGCGGTGAAGCAGATGCAAG
CGACGACTTTGAAGGCACATATGAAGAATGGCGTGAACATATGTGGAGTGACGTAGCAGCCTACTTTAAC
CTCGACATTGAAAACAGTGAAGATAATAAATCTACTCTTTCACTTCAATTTGTCGACAGCGCCGCGGATA
TGCCGCTTGCGAAAATGCACGGTGCGTTTTCAACGAACGTCGTAGCAAGCAAAGAACTTCAACAGCCAGG
CAGTGCACGAAGCACGCGACATCTTGAAATTGAACTTCCAAAAGAAGCTTCTTATCAAGAAGGAGATCAT
TTAGGTGTTATTCCTCGCAACTATGAAGGAATAGTAAACCGTGTAACAGCAAGGTTCGGCCTAGATGCAT
CACAGCAAATCCGTCTGGAAGCAGAAGAAGAAAAATTAGCTCATTTGCCACTCGCTAAAACAGTATCCGT
AGAAGAGCTTCTGCAATACGTGGAGCTTCAAGATCCTGTTACGCGCACGCAGCTTCGCGCAATGGCTGCT
AAAACGGTCTGCCCGCCGCATAAAGTAGAGCTTGAAGCCTTGCTTGAAAAGCAAGCCTACAAAGAACAAG
TGCTGGCAAAACGTTTAACAATGCTTGAACTGCTTGAAAAATACCCGGCGTGTGAAATGAAATTCAGCGA
ATTTATCGCCCTTCTGCCAAGCATACGCCCGCGCTATTACTCGATTTCTTCATCACCTCGTGTCGATGAA
AAACAAGCAAGCATCACGGTCAGCGTTGTCTCAGGAGAAGCGTGGAGCGGATATGGAGAATATAAAGGAA
TTGCGTCGAACTATCTTGCCGAGCTGCAAGAAGGAGATACGATTACGTGCTTTATTTCCACACCGCAGTC
AGAATTTACGCTGCCAAAAGACCCTGAAACGCCGCTTATCATGGTCGGACCGGGAACAGGCGTCGCGCCG
TTTAGAGGCTTTGTGCAGGCGCGCAAACAGCTAAAAGAACAAGGACAGTCACTTGGAGAAGCACATTTAT
ACTTCGGCTGCCGTTCACCTCATGAAGACTATCTGTATCAAGAAGAGCTTGAAAACGCCCAAAGCGAAGG
CATCATTACGCTTCATACCGCTTTTTCTCGCATGCCAAATCAGCCGAAAACATACGTTCAGCACGTAATG
GAACAAGACGGCAAGAAATTGATTGAACTTCTTGATCAAGGAGCGCACTTCTATATTTGCGGAGACGGAA
GCCAAATGGCACCTGCCGTTGAAGCAACGCTTATGAAAAGCTATGCTGACGTTCACCAAGTGAGTGAAGC
AGACGCTCGCTTATGGCTGCAGCAGCTAGAAGAAAAAGGCCGATACGCAAAAGACGTGTGGGCTGGGTAA
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DNA Sequence (Direct Input)

This function allows the users to copy and paste a DNA sequence, which encodes the gene of interest, into the text field.

An example of DNA sequence is provided below:

ATGACAATTAAAGAAATGCCTCAGCCAAAAACGTTTGGAGAGCTTAAAAATTTACCGTTATTAAACACAG
ATAAACCGGTTCAAGCTTTGATGAAAATTGCGGATGAATTAGGAGAAATCTTTAAATTCGAGGCGCCTGG
TCGTGTAACGCGCTACTTATCAAGTCAGCGTCTAATTAAAGAAGCATGCGATGAATCACGCTTTGATAAA
AACTTAAGTCAAGCGCTTAAATTTGTACGTGATTTTGCAGGAGACGGGTTATTTACAAGCTGGACGCATG
AAAAAAATTGGAAAAAAGCGCATAATATCTTACTTCCAAGCTTCAGTCAGCAGGCAATGAAAGGCTATCA
TGCGATGATGGTCGATATCGCCGTGCAGCTTGTTCAAAAGTGGGAGCGTCTAAATGCAGATGAGCATATT
GAAGTACCGGAAGACATGACACGTTTAACGCTTGATACAATTGGTCTTTGCGGCTTTAACTATCGCTTTA
ACAGCTTTTACCGAGATCAGCCTCATCCATTTATTACAAGTATGGTCCGTGCACTGGATGAAGCAATGAA
CAAGCTGCAGCGAGCAAATCCAGACGACCCAGCTTATGATGAAAACAAGCGCCAGTTTCAAGAAGATATC
AAGGTGATGAACGACCTAGTAGATAAAATTATTGCAGATCGCAAAGCAAGCGGTGAACAAAGCGATGATT
TATTAACGCATATGCTAAACGGAAAAGATCCAGAAACGGGTGAGCCGCTTGATGACGAGAACATTCGCTA
TCAAATTATTACATTCTTAATTGCGGGACACGAAACAACAAGTGGTCTTTTATCATTTGCGCTGTATTTC
TTAGTGAAAAATCCACATGTATTACAAAAAGCAGCAGAAGAAGCAGCACGAGTTCTAGTAGATCCTGTTC
CAAGCTACAAACAAGTCAAACAGCTTAAATATGTCGGCATGGTCTTAAACGAAGCGCTGCGCTTATGGCC
AACTGCTCCTGCGTTTTCCCTATATGCAAAAGAAGATACGGTGCTTGGAGGAGAATATCCTTTAGAAAAA
GGCGACGAACTAATGGTTCTGATTCCTCAGCTTCACCGTGATAAAACAATTTGGGGAGACGATGTGGAAG
AGTTCCGTCCAGAGCGTTTTGAAAATCCAAGTGCGATTCCGCAGCATGCGTTTAAACCGTTTGGAAACGG
TCAGCGTGCGTGTATCGGTCAGCAGTTCGCTCTTCATGAAGCAACGCTGGTACTTGGTATGATGCTAAAA
CACTTTGACTTTGAAGATCATACAAACTACGAGCTGGATATTAAAGAAACTTTAACGTTAAAACCTGAAG
GCTTTGTGGTAAAAGCAAAATCGAAAAAAATTCCGCTTGGCGGTATTCCTTCACCTAGCACTGAACAGTC
TGCTAAAAAAGTACGCAAAAAGGCAGAAAACGCTCATAATACGCCGCTGCTTGTGCTATACGGTTCAAAT
ATGGGAACAGCTGAAGGAACGGCGCGTGATTTAGCAGATATTGCAATGAGCAAAGGATTTGCACCGCAGG
TCGCAACGCTTGATTCACACGCCGGAAATCTTCCGCGCGAAGGAGCTGTATTAATTGTAACGGCGTCTTA
TAACGGTCATCCGCCTGATAACGCAAAGCAATTTGTCGACTGGTTAGACCAAGCGTCTGCTGATGAAGTA
AAAGGCGTTCGCTACTCCGTATTTGGATGCGGCGATAAAAACTGGGCTACTACGTATCAAAAAGTGCCTG
CTTTTATCGATGAAACGCTTGCCGCTAAAGGGGCAGAAAACATCGCTGACCGCGGTGAAGCAGATGCAAG
CGACGACTTTGAAGGCACATATGAAGAATGGCGTGAACATATGTGGAGTGACGTAGCAGCCTACTTTAAC
CTCGACATTGAAAACAGTGAAGATAATAAATCTACTCTTTCACTTCAATTTGTCGACAGCGCCGCGGATA
TGCCGCTTGCGAAAATGCACGGTGCGTTTTCAACGAACGTCGTAGCAAGCAAAGAACTTCAACAGCCAGG
CAGTGCACGAAGCACGCGACATCTTGAAATTGAACTTCCAAAAGAAGCTTCTTATCAAGAAGGAGATCAT
TTAGGTGTTATTCCTCGCAACTATGAAGGAATAGTAAACCGTGTAACAGCAAGGTTCGGCCTAGATGCAT
CACAGCAAATCCGTCTGGAAGCAGAAGAAGAAAAATTAGCTCATTTGCCACTCGCTAAAACAGTATCCGT
AGAAGAGCTTCTGCAATACGTGGAGCTTCAAGATCCTGTTACGCGCACGCAGCTTCGCGCAATGGCTGCT
AAAACGGTCTGCCCGCCGCATAAAGTAGAGCTTGAAGCCTTGCTTGAAAAGCAAGCCTACAAAGAACAAG
TGCTGGCAAAACGTTTAACAATGCTTGAACTGCTTGAAAAATACCCGGCGTGTGAAATGAAATTCAGCGA
ATTTATCGCCCTTCTGCCAAGCATACGCCCGCGCTATTACTCGATTTCTTCATCACCTCGTGTCGATGAA
AAACAAGCAAGCATCACGGTCAGCGTTGTCTCAGGAGAAGCGTGGAGCGGATATGGAGAATATAAAGGAA
TTGCGTCGAACTATCTTGCCGAGCTGCAAGAAGGAGATACGATTACGTGCTTTATTTCCACACCGCAGTC
AGAATTTACGCTGCCAAAAGACCCTGAAACGCCGCTTATCATGGTCGGACCGGGAACAGGCGTCGCGCCG
TTTAGAGGCTTTGTGCAGGCGCGCAAACAGCTAAAAGAACAAGGACAGTCACTTGGAGAAGCACATTTAT
ACTTCGGCTGCCGTTCACCTCATGAAGACTATCTGTATCAAGAAGAGCTTGAAAACGCCCAAAGCGAAGG
CATCATTACGCTTCATACCGCTTTTTCTCGCATGCCAAATCAGCCGAAAACATACGTTCAGCACGTAATG
GAACAAGACGGCAAGAAATTGATTGAACTTCTTGATCAAGGAGCGCACTTCTATATTTGCGGAGACGGAA
GCCAAATGGCACCTGCCGTTGAAGCAACGCTTATGAAAAGCTATGCTGACGTTCACCAAGTGAGTGAAGC
AGACGCTCGCTTATGGCTGCAGCAGCTAGAAGAAAAAGGCCGATACGCAAAAGACGTGTGGGCTGGGTAA
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