Table 1. Genetic Modules and Other Vectors in the Fungal Toolkit for Modular Cloning (FTK)a.
| vector name | Addgene ID | part type | unit description | recipient MoClo backbone | template | source and references |
|---|---|---|---|---|---|---|
| pFTK001 | 171273 | P1 | Pact Pc20g11630 promoter | pICH41295 | P. rubens DS54468 | (27) |
| pFTK002 | 171274 | P1 | PgndA An11g02040 promoter | pICH41295 | A. niger N402 | (7), 27 |
| pFTK003 | 171275 | P1 | Patp9 An04g08190 promoter | pICH41295 | A. niger N402 | (27) |
| pFTK004 | 171276 | P1 | PgpdA ANIA_08041 promoter | pICH41295 | pDONR221-AMDS | (27) |
| pFTK005 | 171277 | P1 | Ptef1 ANIA_04218 promoter | pICH41295 | pFC334 (Addgene ID 87846) | (32) |
| pFTK006 | 171278 | P1 | Ptef EF1-subunit ANIA_02063 promoter | pICH41295 | A nidulans FGSC A4 | (33) |
| pFTK007 | 171279 | P1 | PfraA An16g04690 promoter | pICH41295 | A. niger N402 | (5) |
| pFTK008 | 171280 | P1 | Poat1 Pc18g03600 promoter | pICH41295 | P. rubens Wisconsin 54–1255 | (34) |
| pFTK009 | 171281 | P1 | PglaA An03g06550 promoter | pICH41295 | pEBA520 | (5) |
| pFTK010 | 171282 | P1 | PxlnA ANIA_03613 promoter | pICH41295 | A nidulans FGSC A4 | (35) |
| pFTK011 | 171283 | P1 | PpcbAB Pc21g21390 promoter | pICH41295 | P. rubens DS54468 | (27) |
| pFTK012 | 171284 | P1 | P40s An0465 promoter | pICH41295 | pDSM-JAK-108 | (7), 36 |
| pFTK013 | 171285 | P1 | PpcbC Pc21g21380 promoter | pICH41295 | P. rubens DS54468 | (27) |
| pFTK014 | 171286 | P1 | PAnHisB AN6536.2 promoter | pICH41295 | A. niger N402 | (37) |
| pFTK015 | 171287 | P1 | PpyrG AO090011000868 promoter | pICH41295 | pMF21.1 | (38) |
| pFTK016 | 171288 | P1 | 1x TetO UAS + CPgpdA (fused) | pICH41295 | pVG2.2 | (5) |
| pFTK017 | 171289 | P1 | 4x TetO UAS+ CPgpdA (fused) | pICH41295 | pVG2.2 | (5) |
| pFTK018 | 171290 | P1 | 6x TetO UAS + CPgpdA (fused) | pICH41295 | pVG2.2 | (5) |
| pFTK019 | 171291 | P1 | 10x TetO UAS + CPgpdA (fused) | pICH41295 | pVG2.2 | (5) |
| pFTK020 | 171292 | P1 | PalcA synt_NoCrea (ANIA_08979) promoter | pICH41295 | A nidulans FGSC A4 | (39) |
| pFTK021 | 171293 | P1 | Pu3 hom., Putp25, P. rubens Pol-III promoter | pICH41295 | P. rubens DS54468 | (28) |
| pFTK022 | 171294 | P1 | Pu6 hom., P. rubens Pol-III promoter | pICH41295 | P. rubens DS54468 | (28) |
| pFTK023 | 171295 | P1 | PtRNA[Met] P. rubens Pol-III promoter | pICH41295 | P. rubens DS54468 | (28) |
| pFTK024 | 171296 | P1 | PtRNA[Leu] P. rubens Pol-III promoter | pICH41295 | P. rubens DS54468 | (28) |
| pFTK025 | 171297 | P1 | CPpcbC Pc21g21380 (no UAS) core promoter | pICH41295 | P. rubens Wisconsin 54–1255 | (7) |
| pFTK026 | 171298 | P1 | CPnirA AN0098 (no UAS) core promoter | pICH41295 | A nidulans FGSC A4 | (7) |
| pFTK027 | 171299 | P1 | CPura3 YEL021W (no UAS) core promoter | pICH41295 | S. cerevisiae CEN.PK113–7D | (7) |
| pFTK028 | 171300 | P2 | 1xQUAS UAS (for fusion) | pAGM1251 | synthetic DNA | (7), 40 |
| pFTK029 | 171301 | P2 | 5xQUAS UAS (for fusion) | pAGM1251 | synthetic DNA | (7), 40 |
| pFTK030 | 171302 | P2 | 11xQUAS UAS (for fusion) | pAGM1251 | synthetic DNA | (7), 40 |
| pFTK031 | 171303 | P2 | 5xLexA_UAS UAS (for fusion) | pAGM1251 | synthetic DNA | (41) |
| pFTK032 | 171304 | P3 | CPpcbC Pc21g21380 core promoter (for fusion) | pAGM1276 | A nidulans FGSC A4 | (7) |
| pFTK033 | 171305 | P3 | CPnirA AN0098 core promoter (for fusion) | pAGM1276 | S. cerevisiae CEN.PK113-7D | (7) |
| pFTK034 | 171306 | P3 | CPura3 YEL021W core promoter (for fusion) | pAGM1276 | A. niger N402 | (7) |
| pFTK035 | 171307 | CDS1 | ergA Pc22g15550 terbinafine, selection marker | pICH41308 | P. rubens DS54468 | (42) |
| pFTK036 | 171308 | CDS1 | amdS ANIA_08777 acetamidase, selection marker | pICH41308 | pDONR221-AMDS | (29) |
| pFTK037 | 171309 | CDS1 | bleoR phleomycin, selection marker | pICH41308 | pDSM-JAK-109 | (29), 36 |
| pFTK038 | 171310 | CDS1 | hph hygromycin selection marker (hygR) | pICH41308 | pAN7.1 | (43) |
| pFTK039 | 171311 | CDS1 | pyrG AO090011000868 orotidine 5′-phosphate decarboxylase, selection marker | pICH41308 | pMF21.1 | (38) |
| pFTK040 | 171312 | CDS1 | sdh-H85L An14g04400 succinate dehydrogenase, selection marker | pICH41308 | A. niger N402 | (44) |
| pFTK041 | 171313 | CDS1 | hisB Pc20g11690 histidine, selection marker | pICH41308 | P. rubens DS54468 | (37) |
| pFTK042 | 171314 | CDS1 | hisB AN6536.2 histidine, selection marker | pICH41308 | pSE1.6 | (37) |
| pFTK043 | 171315 | CDS1 | eGFP fluorescent reporter | pICH41308 | pLM2_30 (Addgene ID 154222) | (7) |
| pFTK044 | 171316 | CDS1 | eGFP-NLS fluorescent reporter | pICH41308 | pLM2_30 (Addgene ID 154222) | (7) |
| pFTK045 | 171317 | CDS1 | eGFP-SKL fluorescent reporter | pICH41308 | pLM2_30 (Addgene ID 154222) | (7) |
| pFTK046 | 171318 | CDS1 | DsRed.T1 fluorescent reporter | pICH41308 | pDSM-JAK-109 | (7), 27, 36 |
| pFTK047 | 171319 | CDS1 | DsRed-NLS fluorescent reporter | pICH41308 | pDSM-JAK-109 | (7), 27, 36 |
| pFTK048 | 171320 | CDS1 | DsRed.T1-SKL fluorescent reporter | pICH41308 | pDSM-JAK-109 | (27), 36, 45 |
| pFTK049 | 171321 | CDS1 | mCherry fluorescent reporter | pICH41308 | pURA3_1147651 cP_mCherry | (6) |
| pFTK050 | 171322 | CDS1 | dTomato fluorescent reporter | pICH41308 | pMF30.1 | (46) |
| pFTK051 | 171323 | CDS1 | eBFP fluorescent reporter | pICH41308 | pLM2_30 (Addgene ID 154222) with Y66H/Y145F mutations | (7) |
| pFTK052 | 171324 | CDS1 | firefly luciferase reporter | pICH41308 | pVG4.1 | (5) |
| pFTK053 | 171325 | CDS1 | alcR ANIA_08978 transcriptional activator | pICH41308 | A nidulans FGSC A4 | (39) |
| pFTK054 | 171326 | CDS1 | QS (QA-1S) codon optimized, quinic acid repressor | pICH41308 | pAC-Qsco, (Addgene ID 46106) | (40) |
| pFTK055 | 171327 | CDS1 | rtTA2S-M2 (TetR-3xVP16) transcriptional activator | pICH41308 | pVG2.2 | (5) |
| pFTK056 | 171328 | CDS1 | SpCas9-NLS | pICH41308 | pYTK036 (Addgene ID 65143) | (14) |
| pFTK057 | 171329 | CDS1 | dSpCas9(m4)-VPR-NLS | pICH41308 | pYTK036 (Addgene ID 65143), pAG414GPD (Addgene ID 63801) | (14), 47 |
| pFTK058 | 171330 | CDS2 | QF DBD from QA-1F (for fusion) | pICH41258 | pAC-7-QFBDAD (Addgene ID 46096) | (40) |
| pFTK059 | 171331 | CDS2 | LexA DBD (for fusion) | pICH41258 | FRP718_PACT1(-1-520)-LexA-ER-haB42-TCYC1 (Addgene ID 58431) | (48) |
| pFTK060 | 171332 | CDS2 | Gal4D BD (for fusion) | pICH41258 | S. cerevisiae CEN.PK113-7D | (49) |
| pFTK061 | 171333 | CDS2 | SpCas9 (for fusion) | pICH41258 | pYTK036 (Addgene ID 65143) | (14), 21 |
| pFTK062 | 171334 | CDS2 | dSpCas9(m2) (for fusion) | pICH41258 | pYTK036 (Addgene ID 65143) | (14), 21 |
| pFTK063 | 171335 | CDS2 | dSpCas9(m4) (for fusion) | pICH41258 | pYTK036 (Addgene ID 65143) | (14), 21 |
| pFTK064 | 171336 | CDS3 | QF AD from QA-1F (for fusion) | pAGM1299 | pAC-7-QFBDAD (Addgene ID 46096) | (7), 40 |
| pFTK065 | 171337 | CDS3 | B42 AD (for fusion) | pAGM1299 | FRP718_PACT1(-1-520)-LexA-ER-haB42-TCYC1 (Addgene ID 58431) | (48) |
| pFTK066 | 171338 | CDS3 | Gal4 AD (for fusion) | pAGM1299 | S. cerevisiae CEN.PK113-7D | (49) |
| pFTK067 | 171339 | CDS3 | VP16 AD (for fusion) | pAGM1299 | pVG2.2 | (5), 49 |
| pFTK068 | 171340 | CDS3 | VP64 AD (for fusion) | pAGM1299 | pcDNA-dCas9-VP64 (Addgene ID 47107) | (47) |
| pFTK069 | 171341 | CDS4 | eGFP-NLS fluorescent reporter (for fusion) | pAGM1301 | pLM2_30 (Addgene ID 154222) | (7) |
| pFTK070 | 171342 | CDS4 | eYFP-NLS fluorescent reporter (for fusion) | pAGM1301 | pLM2_30 (Addgene ID 154222) with S65G/V68L/S72A/T203Y mutations | (7) |
| pFTK071 | 171343 | CDS5 | eGPF-NLS fluorescent reporter (for fusion) | pICH41264 | PX458 (Addgene ID 48138) | (7) |
| pFTK072 | 171344 | CDS5 | p300core HAT AD, Homo sapiens E1A binding protein p300 (for fusion) | pICH41264 | pcDNA-dCas9-p300 (Addgene ID 61357) | (15), 47, 50 |
| pFTK073 | 171345 | CDS5 | RTT109 HAT AD (for fusion) | pICH41264 | S. cerevisiae CEN.PK113-7 | (51) |
| pFTK074 | 171346 | CDS5 | VPR-NLS AD (for fusion) | pICH41264 | pAG414GPD (Addgene ID 63801) | (47) |
| pFTK075 | 171347 | T1 | TamdS ANIA_08777 terminator | pICH41276 | pDONR221-AMDS | (29) |
| pFTK076 | 171348 | T1 | Ttif35 Pc22g19890 terminator | pICH41276 | pDSM-JAK-108 | (7), 36 |
| pFTK077 | 171349 | T1 | TpenDE Pc21g21370 terminator | pICH41276 | P. rubens Wisconsin 54–1255 | (36) |
| pFTK078 | 171350 | T1 | TxlnA ANIA_03613 terminator | pICH41276 | A nidulans FGSC A4 | (35) |
| pFTK079 | 171351 | T1 | Toat1 Pc18g03600 terminator | pICH41276 | P. rubens Wisconsin 54–1255 | (34) |
| pFTK080 | 171352 | T1 | Tcyc1 YJR048W terminator | pICH41276 | pDSM-JAK-109 | (36) |
| pFTK081 | 171353 | T1 | TactA (Tact1) ANIA_06542 P. rubens terminator | pICH41276 | pDSM-JAK-108 | (7), 36 |
| pFTK082 | 171354 | T1 | Tu3 hom., Tutp25, P. rubens Pol-III terminator | pICH41276 | P. rubens DS54468 | (28) |
| pFTK083 | 171355 | T1 | Tu6 hom., P. rubens Pol-III terminator | pICH41276 | P. rubens DS54468 | (28) |
| pFTK084 | 171356 | T1 | TtRNA[Met] A. niger Pol-III terminator | pICH41276 | A. niger N402 | (28) |
| pFTK085 | 171357 | T1 | TtRNA[Met] A. niger Pol-III terminator | pICH41276 | A. niger N402 | (28) |
| pFTK086 | 171358 | TU | P-ANtRNA[Arg21]-sgRNA-dummy-Esp3I, Pol-III sgRNA transcription unit | pICH41331 | A. niger N402 | (31) |
| pFTK087 | 171359 | TU | P-ANtRNA[Pro1]-sgRNA-dummy-Esp3I, Pol-III sgRNA transcription unit | pICH41331 | A. niger N402 | (31) |
| pFTK088 | 171360 | TU | AMA1 sequence (short), entry vector providing fungal replication | pICH41331 | pDSM-JAK-109 | (25), 36 |
| pFTK089 | 171361 | TU | penicillin gene cluster P. rubens 5′ flanking region | pICH41331 | P. rubens DS54468 | (7) |
| pFTK090 | 171362 | TU | penicillin gene cluster P. rubens 3′ flanking region | pICH41331 | P. rubens DS54468 | (7) |
| pFTK091 | 171363 | TU (level 1) | pks17 Pc21g16000 (conidial pigment biosynthesis) P. rubens 5′ flanking region | pICH47732 (lvl1) | P. rubens DS54468 | (28) |
| pFTK092 | 171364 | TU (level 1) | pks17 Pc21g16000 (conidial pigment biosynthesis) P. rubens 3′ flanking region | pICH47772 (lvl1) | P. rubens DS54468 | (28) |
| pFTK093 | 171365 | TU (level 1) | sgRNA transcription unit (MoClo lvl1 unit), P-gpdA-HH-sgRNA-HDV-Ttrpc | pICH47761 (lvl1) | pFC334 (Addgene ID 87846), pLM-AMA18.0 dCas9-VPR (Addgene ID 138945) | (14) |
| pFTK094-LM-AMA002.0 | 171366 | AMA1 | pLM-AMA002, fungal shuttle vector with bleoR marker for MoClo entry vectors | n/a | pDSM-JAK-109 | (36) |
| pFTK095-LM-AMA015.0 | 171367 | AMA1 | pLM-AMA15.0, CRISPR/Cas9 genome editing with HH-sgRNA-HDV transcription unit, ergA and bleoR fungal markers | n/a | pDSM-JAK-109, pYTK036 (Addgene ID 65143), pLM-AMA15.0 Cas9 (Addgene ID 138944) | (14) |
| pFTK096-LM-AMA018.0 | 171368 | AMA1 | pLM-AMA18.0, CRISPRa/dSpCas9-VPR transcriptional activator with HH-sgRNA-HDV transcription unit, ergA and bleoR fungal markers | n/a | pDSM-JAK-109, pYTK036 (Addgene ID 65143), pAG414GPD (Addgene ID 63801), pLM-AMA18.0 dCas9-VPR (Addgene ID 138945) | (14) |
a
Units in the toolkit are described using a vector name, an Addgene ID, a part type specifying the function of the part (P, promoter; CDS, coding sequence; T, terminator; TU, transcription unit; AMA1, AMA1-sequence-based fungal replicating vector), a short description of the vector, its recipient Modular Cloning destination vector, the source of the genetic element, and its applications(s).