Table 1. Original eGFP transposon insertion sites within the human CLN3 peptide.
clone ID | transposon after a.a. | Peptide sequence |
1 | 36 | HWK _transposon_HWKNAV |
2 | 85 | PHN_transposon_PHNSSS |
3 | 101 | AVL_transposon_AVLLAD |
4 | 125 | PYS_transposon_PYSPRV |
5 | 150 | VGT_transposon_VGTSLC |
6 | 203 | GLT_transposon_GLTQAG |
7 | 238 | AQD_transposon_AQDPGG |
8 | 279 | VFK_transposon_VFKGLL |
9 | 345 | RFT_transposon_RFTWAL |
10 | 348 | WAL_transposon_WALALL |
11 | 406 | HRE_transposon_HREFAM |
The transposon encoding for eGFP was inserted into the pCMV5 expression vector containing the cDNA for the full-length human CLN3 amino acid sequence at indicated positions by transposase enzyme. The clone # is a running number for different clones as of the N-terminus, (left column), the middle column shows the number of the amino acid in the CLN3 peptide sequence after which the transposon was inserted, and the right column shows the corresponding hCLN3 amino acid sequence. eGFP in clones 1 to 10 was later replaced with a myc-tag and combined with hCLN3-eGFP11 to create chimeric clones with a near C-terminal eGFP and one myc-tag further towards the N-terminus. Refer to Figure 4A for visual location along the previously published model for hCLN3. Note that the last 3 amino acids before transposon insertion are duplicated behind the transposon insertion site (see Figure 1).