LABORATORY OF BIOMOLECULAR ENGINEERING AND NANOMEDICINE
Principal Investigator: Gang Bao
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TAL DESIGN & ANALYSIS GROUP
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TAL Plasmids Sequence Assembly Tool
        This tool is designed to aid in sequence confirmation of Transcription Activator-Like (TAL) plasmids assembled using a variety of popular methods.

        Cermak et al. (PMID: 21493687), also known as Golden Gate Assembly or the Voytas Toolkit.
        ----FASTA formatted plasmid sequences for pFUS constructs as well as final destination vectors can be generated.
        ----Genbank/ApE formatted plasmid maps can be generated for final destination vectors.
        ----Sequencing reads can be aligned for pFUS constructs and destination vectors using the Quick Align Tool.
        ----An online video tutorial is available for using the tool with this assembly method.

        Sanjana et al. (PMID: 22222791), the PCR based Golden Gate method.
        ----Using the plasmid templates generated by TALEffectors.com, sequencing reads can be aligned using the Quick Align Tool.
        ----An online video tutorial is available for using the tool with this assembly method.

    Flag Counter
        Reyon & Tsai et al. (PMID: 22484455), also known as FLASH assembly.
        ----FASTA formatted plasmid sequences for the TALEN vectors can be generated.
        ----Genbank/ApE formatted plasmid maps can be generated (in progress).
        ----Sequencing reads can be aligned to plasmids using the Quick Align Tool.
        ----A worklist file for a liquid handling robot (or a dedicated researcher) to assemble up to 96 TALs can be generated. See Plate Layout for additional details.
        ----An online video tutorial is available for using the tool with this assembly method.

        Briggs et al. (doi:10.1093/nar/gks624), also known as Iterative Capped Assembly (ICA).
        ----FASTA formatted plasmid sequences for TALEN vectors can be generated.
        ----Genbank/ApE formatted plasmid maps can be generated for TALEN vectors.
        ----Sequencing reads can be aligned to plasmids using the Quick Align Tool.

        Recent Updates:
        ----03/19/14:   pZHY500 and pZHY501 plant destination vectors added
        ----12/16/13:   Bao Lab Golden Gate Compatible plasmids now available through Addgene



Generate Plasmid Sequences
Choose Entry Format:
Enter RVDs for TAL(s):
           Enter desired TAL target(s) in FASTA format:
          
Choose Assembly Method:
Cermak et al. (Golden Gate) Reyon et al. (FLASH) Briggs et al. (ICA)
Destination Vector:
Destination Vector: Destination Vector:
More information on Destination Vectors
Default Guanosine RVD: NK NN NH (Voytas Codon) NH (Bao Codon)
Generate ONLY final construct:

Worklist well format:
Alphanumeric (A1, B2, etc.)
Numbers (1, 13, etc.)
 
FASTA Plasmid Constructs:

 Genbank/ApE Final Construct(s):

 Liquid Handler Worklist (paste as text into excel):
Quick Alignment Tool -- Generate alignments of sequencing reads to individual vectors:
Cermak et al.
(Golden Gate)
Vector: Entry Format: Target DNA Sequence:
RVDs:
Forward Sequencing Read: Reverse Sequencing Read:    
Sanjana et al.
(PCR Based)
Enter the template DNA sequence as provided by TALEffectors:
Forward Read:Middle Read:Reverse Read:
Reyon et al.
(FLASH)
Vector: Entry Format: Target DNA Sequence:
RVDs:
Forward Read (oSQT1):Middle Read (oSQT3):Reverse Read (oJS2980):
Briggs et al.
(ICA)
Vector: Entry Format: Target DNA Sequence:
RVDs:
Forward Read:Middle Read:Reverse Read:

Sequence Alignment:
RVD_Locations:
Template_Sequence:
Merged_Reads:
Forward_Read:
Reverse_Read:
Middle_Read:
Destination Vectors Information:
Assembly MethodPlasmid NameDescriptionAddgene
Cermak et al. pTAL1TALE with natural bacterial activator domain31031
pTAL2TALE with natural bacterial activator domain and no stop codon31033
pTAL3TALEN with HIS3 marker31034
pTAL4TALEN with LEU2 marker31035
SangamoMammalian expression TALEN with 3x FLAG tag and (-152, +63) backbone40788
pTALE64GMammalian expression TALE with 3x FLAG tag, (-152, +63) backbone, VP64 activation domain, T2A sequence, and eGFP35188
pTALE64CMammalian expression TALE with 3x FLAG tag, (-152, +63) backbone, VP64 activation domain, T2A sequence, and AmCyanComing Soon
pTALE64Mammalian expression TALE with 3x FLAG tag, (-152, +63) backbone, VP64 activation domain, no fluorescent tag35189
pET-HIS-SangamoE. coli expression TALEN with N-terminal 6xHis tag, 3xFLAG tag, (-152, +63) backbone40786
pET-Sangamo-HISE. coli expression TALEN with 3xFLAG tag, (-152, +63) backbone, C-terminal 6xHis tag40787
pCS2TAL3-DD3xFLAG tag, TALEN with obligate heterodimer DD FokI domain37275
pCS2TALE3-RRTALEN with obligate heterodimer RR FokI domain37276
pTAL5-BBYeast expression TAL repressor with PGAL1 Promoter36033
pTAL6-BBYeast expression TAL repressor with PTEF1 Promoter36034
pCAG-T7-TALEN(Sangamo)-DestinationMammalian expression TALEN with Sangamo backbone37184
pCAG-T7-TALEN(Sangamo)-FokI-ELDMammalian expression TALEN with Sangamo backbone, ELD Heterodimeric FokI variant with Sharkey mutations40132
pCAG-T7-TALEN(Sangamo)-FokI-KKRMammalian expression TALEN with Sangamo backbone, KKR Heterodimeric FokI variant with Sharkey mutations40131
pZHY500Yeast experssion TALEN with Sangamo backbone and HIS3 marker (similar to pTAL3)36186
pZHY501Yeast experssion TALEN with Sangamo backbone and LEU2 marker (similar to pTAL4)36186
RCIscript-GoldyTALENmRNA transcription vector, (-152, +63) backbone, wild-type FokI38142
Reyon et al. JDS70, 71, 74, 78Mammalian expression TALENs with 3x FLAG tag and Sangamo backbone with different C-terminal RVDsTALENkit
Briggs et al. TALENMammalian expression TALENUnknown
The TAL Plasmids Sequence Assembly Tool was created by Eli Fine
If you have comments or would like to see additional publicly available, compatible destination vectors be added, please email EliFine@gatech.edu
If you are interested in receiving updates about the online tools created by the TAL Design & Analysis Group, you can subscribe to our mailing list