Software error:
Global symbol "$groups" requires explicit package name at /websites/parts.igem.org/cgi/lib/User.pm line 84. Global symbol "$groups" requires explicit package name at /websites/parts.igem.org/cgi/lib/User.pm line 84. Global symbol "$groups" requires explicit package name at /websites/parts.igem.org/cgi/lib/User.pm line 85. Compilation failed in require at /websites/parts.igem.org/cgi/lib/Change.pm line 11. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/Change.pm line 11. Compilation failed in require at /websites/parts.igem.org/cgi/lib/Part.pm line 12. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/Part.pm line 12. Compilation failed in require at /websites/parts.igem.org/cgi/lib/IconBar.pm line 10. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/IconBar.pm line 10. Compilation failed in require at /websites/parts.igem.org/cgi/lib/BBWeb.pm line 9. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/BBWeb.pm line 9. Compilation failed in require at /websites/parts.igem.org/cgi/lib/WrapPart.pm line 8. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/WrapPart.pm line 8. Compilation failed in require at /websites/parts.igem.org/cgi/extensions/wiki_header_hook.cgi line 9. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/extensions/wiki_header_hook.cgi line 9.
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Status: 500
Content-type: text/html
Software error:
Global symbol "$groups" requires explicit package name at /websites/parts.igem.org/cgi/lib/User.pm line 84. Global symbol "$groups" requires explicit package name at /websites/parts.igem.org/cgi/lib/User.pm line 84. Global symbol "$groups" requires explicit package name at /websites/parts.igem.org/cgi/lib/User.pm line 85. Compilation failed in require at /websites/parts.igem.org/cgi/lib/IconBar.pm line 12. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/IconBar.pm line 12. Compilation failed in require at /websites/parts.igem.org/cgi/lib/BBWeb.pm line 9. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/BBWeb.pm line 9. Compilation failed in require at /websites/parts.igem.org/cgi/lib/Change.pm line 8. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/Change.pm line 8. Compilation failed in require at /websites/parts.igem.org/cgi/lib/Part.pm line 12. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/Part.pm line 12. Compilation failed in require at /websites/parts.igem.org/cgi/partsdb/putout.cgi line 8. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/partsdb/putout.cgi line 8.
For help, please send mail to this site's webmaster, giving this error message and the time and date of the error.
Status: 500
Content-type: text/html
Software error:
Global symbol "$groups" requires explicit package name at /websites/parts.igem.org/cgi/lib/User.pm line 84. Global symbol "$groups" requires explicit package name at /websites/parts.igem.org/cgi/lib/User.pm line 84. Global symbol "$groups" requires explicit package name at /websites/parts.igem.org/cgi/lib/User.pm line 85. Compilation failed in require at /websites/parts.igem.org/cgi/lib/IconBar.pm line 12. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/IconBar.pm line 12. Compilation failed in require at /websites/parts.igem.org/cgi/lib/BBWeb.pm line 9. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/BBWeb.pm line 9. Compilation failed in require at /websites/parts.igem.org/cgi/lib/Change.pm line 8. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/Change.pm line 8. Compilation failed in require at /websites/parts.igem.org/cgi/lib/Part.pm line 12. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/lib/Part.pm line 12. Compilation failed in require at /websites/parts.igem.org/cgi/partsdb/putout.cgi line 8. BEGIN failed--compilation aborted at /websites/parts.igem.org/cgi/partsdb/putout.cgi line 8.
For help, please send mail to this site's webmaster, giving this error message and the time and date of the error.
Design Notes
Addition of a protein coding domain to the suffix by Standard Assembly RFC[10] creates a scar-site. As the scarsite encodes a stop codon (uacuag), this will leave the fused protein coding domain untranslated.
Instead we recommend the following assembly method:
1) Design primers for amplification of the C-terminal fusion protein with BamHI resitriction site included in the forward primer. The primers must also include prefix and suffix.
1.2) Forward primer: 5'-ATATGGATCCNNN...NNN-3'
1.3) Reverse primer: 5'-ATATCTGCAGCGGCCGCTACTAGTANNN..NNN-3'
2) Amplify through PCR with designed primers
3) Digest PCR-product and pSB1C3-T18 with BamHI and PstI.
4) Ligate the two digested product.
Source
pUT18C