LIC Generator, a resource to generate primers for Ligation Independent Cloning (LIC)

This program generates the sequence of a set of 4 primers to amplify both insert and vector for LIC cloning.
Easily introduce tags or protease cleavage sites in your vector/insert.
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LIC requires only three steps Compact display

1- Choose your LIC1 sequence (5' to your insert)

check one or several boxes
OptionVector partLIC1Insert partdNTP
(insert)
RemarksLinker*
Shine-DalgarnoCTAAGGAGGTTAATCATGdGTPNo extra sequence
KozakAGGGCCGCCGCCACCATGdTTPNo extra sequence
KozakTAAGGAGGCCACCATGdATPNo extra sequence
KozakGAAACCACCACCATGdCTPNo extra sequence
Simple LICACCATGGGTGGTGGTGGCdGTPA at -3
Simple LICACCATGGGTGGTGGTGGCATGdGTPATG conserved
Simple LICACCATGAGCGGCGCAATGdATPATG conserved
Simple LICACCATGAGCGGCGCAGTGdATPA at -3
Simple LICACCATGTCGGGCGGCATGdTTPATG conserved
Simple LICACCATGTCGGGCGGCAdTTPA at -3
Simple LICACCATGACCCTCTCCCTTTCGdCTPA at -3
Simple LICACCATGACCCTCTCCCTTTCGATGdCTPATG conserved
His TagATGCATCACCACCACCACCACGGTdCTPNsiI
His TagACCATGAGCGCGCATCACCACCACCACCACGGTdCTPA at -3Spacer
His TagACCATGCATCACCACCACCACCACGGTdCTPA at -3
His TagGCGCATCACCACCACCACCACGGTdCTPNo ATG
His TagAGCGCGCATCACCACCACCACCACGGTdCTPNo ATGSpacer
His TagATGCATCACCACCACCACCACGGTATGdCTPATG conserved
His TagACCATGAGCGCGCATCACCACCACCACCACGGTATGdCTPA at -3Spacer
His TagATGCATCACCACCACCACCACTCTdATPNsiI
His TagACCATGAGCGCGCATCACCACCACCACCACTCTdATPA at -3Spacer
His TagATGAGCGCGCATCACCACCACCACCACTCTdATPSpacer
His TagAGCGCGCATCACCACCACCACCACTCTdATPNo ATGSpacer
His TagCATCACCACCACCACCACTCTATGdATP
His TagACCATGAGCGCGCATCACCACCACCACCACTCTATGdATPA at -3Spacer
His TagCATCACCACCACCACCACTCTdATPNo ATG
Strep TagACCATGAGCGCTTGGAGCCACCCACAATTCGAGAAGdCTPA at -3Spacer
Strep TagACCATGTGGAGCCACCCACAATTCGAGAAGdCTPA at -3
Strep TagACCATGAGCGCTTGGAGCCACCCACAATTCGAGAAGATGdCTPATG conservedSpacer
Strep TagACCATGTGGAGCCACCCACAATTCGAGAAGATGdCTPATG conserved
Strep TagACCATGAGCGCTTGGAGCCACCCGCAGTTCGAGAAGdATPA at -3Spacer
Strep TagACCATGTGGAGCCACCCGCAGTTCGAGAAGdATPA at -3
Strep TagACCATGAGCGCTTGGAGCCACCCGCAGTTCGAGAAGdATPA at -3Spacer
Strep TagACCATGTGGAGCCACCCGCAGTTCGAGAAGdATPA at -3
Strep TagAGCGCTTGGAGCCACCCGCAGTTCGAGAAGdATPNo ATGSpacer
Strep TagTGGAGCCACCCGCAGTTCGAGAAGdATPNo ATG
TevAGCGCTGAGAACTTATACTTCCAAAGCTTCdCTPNo peptidaseSpacer
TevGAGAACTTATACTTCCAAAGCTTCdCTPNo peptidase
TevAGCGCTGAGAACTTATACTTCCAAAGCdCTPSpacer
TevGAGAACTTATACTTCCAAAGCdCTP
PrescissionGCCGGTTTGGAGGTGCTCTTCCAGGGTCCCdGTP
PrescissionAGCGCTGCCGGTTTGGAGGTGCTCTTCCAGGGTCCCdGTPSpacer
PrescissionGCCGGTTTGGAGGTGCTCTTCCAGGGTCCCATGdGTPATG conserved
PrescissionAGCGCTGCCGGTTTGGAGGTGCTCTTCCAGGGTCCCATGdGTPATG conservedSpacer
ThrombinAACGCTCTGGTGCCACGTdTTP
Factor XaAGCGCTGGCGGTATTGAGGGTCGCdGTPSpacer
Factor XaGGCGGTATTGAGGGTCGCdGTP
Factor XaGGCGGTATTGAGGGTCGCATGdGTPATG conserved
EnterokinaseAGCGCTGGTGACGACGACAAGGGTdATPSpacer
EnterokinaseAGCGCTGGTGACGACGACAAGATGdATPATG conservedSpacer
EnterokinaseGGTGACGACGACAAGGGTdATP
EnterokinaseGGTGACGACGACAAGATGdATPATG conserved


2- Choose your LIC2 sequence (3' from your insert)

check one or several boxes

Remark: LIC1 and LIC2 must match with respect to the nucleotide (same color code)

ChoiceInsert partLIC2Vector partdNTP
(insert)
RemarksLinker*
Simple LICTAGCGTTGTGAGGGTGCdCTP
Simple LICAGCGGTGGTGGTGGCTAGdCTPextra sequence
Simple LICTGAGTTGTGCGGGTGAdATP
Simple LICTAGCCTAACCTAACCCGdGTP
Simple LICTAGGCAGCAAGCCTdTTP
His TagATGCATCACCACCACCACCACGGTdGTPStop missing
His TagGTGCACCACCACCACCACCACGGTdGTPStop missing
His TagAGCGCGATGCATCACCACCACCACCACGGTdGTPStop missingSpacer
His TagAGCGCGGTGCACCACCACCACCACCACGGTdGTPStop missingSpacer
His TagATGCATCACCACCACCACCACGGTTAGdGTPNsiI in tag
His TagGTGCACCACCACCACCACCACGGTTAGdGTPSnoI in tag
His TagATGCATCACCACCACCACCACTCTdTTPStop missing
His TagCATCACCACCACCACCACTCTdTTPStop missing
His TagAGCGCGCATCACCACCACCACCACTCTdTTPStop missingSpacer
His TagAGCGCGCATCACCACCACCACCACTCTdTTPStop missingSpacer
His TagATGCATCACCACCACCACCACTAGdTTPNsiI in tag
His TagCATCACCACCACCACCACTAGdTTPNoATG but Stop
Strep TagAGCGCTTGGAGCCACCCACAATTCGAGAAGTAGdGTPSpacer
Strep TagTGGAGCCACCCACAATTCGAGAAGTAGdGTP
Strep TagAGCGCTTGGAGCCACCCACAATTCGAGAAGdGTPStop missingSpacer
Strep TagTGGAGCCACCCACAATTCGAGAAGdGTPStop missing
Strep TagAGCGCTTGGAGCCACCCGCAGTTCGAGAAGTAGdTTPSpacer
Strep TagTGGAGCCACCCGCAGTTCGAGAAGTAGdTTP
Strep TagAGCGCTTGGAGCCACCCGCAGTTCGAGAAGdTTPStop missingSpacer
Strep TagTGGAGCCACCCGCAGTTCGAGAAGdTTPStop missing
TevAGCGCTGAGAACTTATACTTCCAAAGCGCGdGTPSpacer
TevAGCGCTGAGAACTTATACTTCCAAAGCGCGdGTPSpacer
TevGAGAACTTATACTTCCAAAGCdGTP
TevAGCGCTGAGAACTTATACTTCCAAAGCTTCdGTPNo peptidaseSpacer
TevGAGAACTTATACTTCCAAAGCTTCdGTPNo peptidase
PrescissionAGCGCTGCCGGTTTGGAGGTGCTCTTCCAGGGTCCCdCTPSpacer
PrescissionGCCGGTTTGGAGGTGCTCTTCCAGGGTCCCdCTP
ThrombinAACGCTCTGGTGCCACGTdATP
Factor XaAGCGCTGGCGGTATTGAGGGTCGCdCTPSpacer
Factor XaGGCGGTATTGAGGGTCGCdCTP
EnterokinaseAGCGCTGGTGACGACGACAAGGGTdTTPSpacer
EnterokinaseAGCGCTGGTGACGACGACAAGATGdTTPATG after EKSpacer
EnterokinaseGGTGACGACGACAAGGGTdTTP
EnterokinaseGGTGACGACGACAAGATGdTTPATG after EK
EnterokinaseAGCGCTGGCGATGATGATAAGGGCdCTPSpacer
EnterokinaseAGCGCTGGCGATGATGATAAGATGGGCdCTPATG after EKSpacer
EnterokinaseGGCGATGATGATAAGGGCdCTP
EnterokinaseGGCGATGATGATAAGATGGGCdCTPATG after EK
*Kozak means that a Kozak consensus is present before the Start
*Linker means that a Ser-Ala linker was added for example between the initiation Met and the tag


3- Add the informations about your project below

(not recorded)
Enter the sequence of your insert (or at least the extremities comprising ATG and Stop codon) plain text, numbers and spaces will be removed
Demo sequence
Enter a name for your project (recommended)
Enter the sequence of your vector preceding the insert (30 signs min, 100 signs max, plain text)
Enter the sequence of your vector following the insert (30 signs min, 100 signs max, plain text)


More about LIC ? | Protocols | Contact: marie-claire.dagher(at)ujf(dash)grenoble(dot)fr
Marie-Claire Dagher - UMR CNRS 5525, TIMC-IMAG Equipe TheREx logo cnrs    logo Universite Joseph Fourier  Acknowledgements:  logo cea