Multisite Assembly of Gateway Induced Clones (MAGIC): a flexible cloning toolbox with diverse applications in vertebrate model systems.
Gillespie W, Zhang Y, Ruiz OE, Cerda J, Ortiz-Guzman J, Turner WD, Largoza G, Sherman M, Mosser LE, Fujimoto E, Chien CB, Kwan KM, Arenkiel BR, Devine WP, Wythe JD
bioRxiv [Preprint]. 2024 Jul 13:2024.07.13.603267. doi: 10.1101/2024.07.13.603267.
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Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 224472 | p5E-CAGGS (JDW 912) | Gateway compatible 5' entry clone containing the CAG promoter for constitutive expression of downstream constructs. |
| 224473 | p5E-EF1a (JDW 1164) | Gateway compatible 5' entry clone containing the human EF1a promoter |
| 224474 | p5E-EFS (EF1a core) (JDW 1319) | Gateway compatible 5' entry clone with human alpha 2 globin pause site (to prevent transcription readthrough) and EF1a core promoter. Lacks downstream intron for a lower-activity, compact promoter. |
| 224475 | p5E-TetO8x-CMVmin (JDW 1087) | Gateway compatible 5' entry clone containing 8 tet operons (TREs) upstream of a minimal CMV promoter |
| 224476 | p5E TRE-3GV (JDW 1163) | Gateway compatible 5' entry clone containing a 3rd generation Tet-On promoter |
| 224477 | p5E lexOP-c-fos min pro (JDW 1254) | Gateway compatible 5' entry clone containing a Lex Operon upstream of a c-fos minimal promoter |
| 224478 | p5E-mmCdh5pro (JDW 1239) | Gateway compatible 5' entry clone containing the Murine Cdh5 promoter |
| 224479 | p5E-hs_ICAM_full (JDW 1047) | Gateway compatible 5' entry clone containing the Human ICAM2 promoter |
| 224480 | p5E-hs_ICAM_short (JDW 1046) | Gateway compatible 5' entry clone containing the Human minimal ICAM2 promoter |
| 224481 | p5E-hs_Cldn5 (JDW 1120) | Gateway compatible 5' entry clone containing the Human Claudin 5 promoter |
| 224482 | p5E-dr_Unc (JDW 1365) | Gateway compatible 5' entry clone containing the Zebrafish unc-503 promoter with a minimal c-fos promoter and b-globin intron |
| 224483 | p5E_hs_Glast (JDW 1182) | Gateway compatible 5' entry clone containing the Human GLAST promoter |
| 224484 | pME V5-mTagBFP2 (JDW 830) | Gateway compatible middle entry clone containing V5 tagged mTagBFP2 (Cytosolic blue fluorescent reporter) |
| 224485 | pME AmCyan (JDW 1151) | Gateway compatible middle entry clone containing AmCyan cytosolic blue flourescent reporter |
| 224486 | pME-H2B-V5-n2StayGoldC4 (JDW 1354) | Gateway compatible middle entry clone contianing Histone H2B fusion to V5-tagged (n2)StayGold(C4) (Nuclear StayGold fluorescent reporter) |
| 224487 | pME V5 mScarlet-I (JDW 1323) | Gateway compatible middle entry clone containing V5 tagged m-Scarlet-I (Cytosolic far red fluorescent reporter) |
| 224488 | pME-Luciferase-P2A-H2A-mCherry (JDW 926) | Gateway compatible middle entry clone containing a Luciferase P2A H2A-mCherry dual reporter (Luciferase/nuclear red fluorescent reporter) |
| 224489 | pME-Sun1-2xsfGFP-6xMYC-pA (JDW 681) | Gateway compatible middle entry clone containing Sun1 reporter fusion to 2 copies of sfGFP and 6 MYC tags (Nuclear envelope reporter and affinity tag) |
| 224490 | pME_Golgi-BFP_P2A_H2A_iRFP (JDW 1007) | Gateway compatible middle entry clone containing GalT-mTagBFP-HA-P2A-H2A-iRFP (BFP golgi reporter and infrared RFP nuclear reporter) |
| 224491 | pME_Golgi-BFP_P2A_H2A_tdiRFP (JDW 1079) | Gateway compatible middle entry clone containing GalT-mTagBFP-HA-P2A-H2A-tdiRFP (BFP golgi reporter and tandem infrared RFP nuclear reporter) |
| 224492 | pME-myr-mTagBFP2-Flag (JDW 1183) | Gateway compatible middle entry clone containing a Myristoylated mTagBP2 with a c-terminal Flag tag (cell membrane BFP marker) |
| 224493 | pME-Lifeact-mCherry-3xHA (JDW 1321) | Gateway compatible middle entry clone containing Lifeact-mCherry (RFP F-actin reporter) |
| 224494 | pME-Lifeact-EGFP-3xHA (JDW 1322) | Gateway compatible middle entry clone containing Lifeact-EGFP (EGFP F-actin reporter) |
| 224495 | pME Actin-Vhh sfGFP (JDW 1215) | Gateway compatible middle entry clone with Actin nanobody fused to sfGFP (GFP actin reporter) |
| 224496 | pME-Actin-Vhh-sfGFP-P2A-1xHA-iRFP-caax (JDW 1216) | Gateway compatible middle entry clone containing an Actin nanobody fused to sfGFP followed by P2A and iRFP-caax tag (GFP actin reporter and cell membrane iRFP reporter) |
| 224497 | pME-Actin-Vhh-sfGFP-P2A-1xHA-tdiRFP-caax (JDW 1311) | Gateway compatible middle entry clone containing an Actin nanobody fused to sfGFP followed by P2A and tdiRFP-caax tag (GFP actin reporter and cell membrane iRFP reporter) |
| 224498 | pME Actin Vhh Halo Tag (JDW 1223) | Gateway compatible middle entry clone containing an Actin nanonbody fused to a flexible linker and Halo Tag (For visualizing actin, actin chromobody) |
| 224499 | pME Actin Vhh mCherry (JDW 1240) | Gateway compatible middle entry clone containing an Actin nanobody fused to mCherry (For visualizing actin, actin chromobody) |
| 224500 | pME Actin Vhh mNeonGreen-HA-pA (JDW 1248) | Gateway compatible middle entry clone containing Actin nanobody fused to mNeonGreen with an HA tag (Visualizing actin, actin chromobody) |
| 224501 | pME Lamin B Vhh mNeonGreen-pA (JDW 1312) | Gateway compatible middle entry clone containing Lamin B nanobody fused to mNeonGreen with HA Tag (For visualizing the nuclear lamina, lamin chromobody) |
| 224502 | pME-mScarlet3-Giantin (JDW 1356) | Gateway compatible middle entry clone containing a V5-tagged mScarlet fused to Giantin (For visualizing the Golgi) |
| 224503 | pME-mCerulean3-TUBB1A (JDW 1367) | Gateway compatible middle entry clone containing mCerulean3 fused to TUBB1A for visualizing tubulin in cells |
| 224504 | pME-LexA-mODC (no LexOp) (JDW 937) | Gateway compatible middle entry clone containing a destabilized LexA transactivator |
| 224505 | pME-LexA-mODC-LexOP-35SCaMV (JDW 1348) | Gateway compatible middle entry clone containing a Destablized LexA and LexOperon with minimal CaMV 35S promoter as an all in one casette for RU486 driven gene expression |
| 224506 | pME-LexA-mODC-LexOP-c-Fos (JDW 1359) | Gateway compatible middle entry clone containing a Destablized LexA and LexOperon with 3' minimal c-Fos promoter for an all in one RU486 driven gene expression construct |
| 224507 | pME TAEL2.0-5xC120-minpro (JDW 1109) | Gateway compatible middle entry clone containing TAEL2.0 transcription factor and 5xC120 response element (All in one binary TAEL and effector domain clone) |
| 224508 | pME-TAEL (JDW 1324) | Gateway compatible middle entry clone containing TAEL2.0 transcription factor for making binary TAEL system vectors (generating a tissue specific driver) |
| 224509 | pME-myc-BirA (JDW 1210) | Gateway compatible middle entry clone containing a Myc tagged BirA |
| 224510 | pME-Myc-BioID2-MCS1 (JDW 1095) | Gateway compatible middle entry clone containing a Myc-tagged BIOID2 construct |
| 224511 | pME-Myc-BioID2-MCS2 (JDW 1209) | A Gateway compatible middle clone with a Myc-tagged BIOID2 construct followed by a flexible linker and an MCS |
| 224512 | pME-3xHA-BirA-T2A-mCherry-caax (JDW 889) | A Gateway compatible middle entry clone containing Biotin Ligase followed by T2A and mCherry membrane reporter (biotinylation and flourescent cell membrane reporter) |
| 224513 | pME-3x-FLAG-Ubiquitin (JDW 1098) | A Gateway compatible middle entry clone containing a 3x FLAG tagged human Ubiquitin for monitoring ubiquitylation status |
| 224514 | pME-3xNLS-(G2SG2)2-Cre (JDW 1149) | A Gateway compatible middle entry clone containing a 3x nuclear localization signal followed by a multiple cloning site and a flexible linker fused to Cre |
| 224515 | pME nls EGFP-Cre (JDW 1329) | A Gateway compatible middle entry clone containing a NLS-EGFP-Cre fusion |
| 224516 | pME-mScarlet-I-P2A-iCre (JDW 917) | A Gateway-compatible middle entry clone containing mScarlet-I P2A-iCre (codon optimized) |
| 224517 | pME-3xNLS-mTagBFP2-Cre (JDW 918) | A Gateway-compatible middle entry clone containing a 3xNLS-mTagBFP2-Cre fusion |
| 224518 | pME-mTurquoise-P2A-iCre (JDW 920) | A Gateway compatible middle entry clone containing mTurquoise P2A iCre (codon optimized Cre) |
| 224519 | pME-EGFP-P2A-iCre (JDW 921) | A Gateway compatible middle entry clone containing EGFP P2A iCre (codon optimized cre) |
| 224520 | pME iCre-I (JDW 1203) | A Gateway compatible middle entry clone containing a loxP flanked, self inactivating Cre that contains an intron to prevent recombination in bacteria |
| 224521 | pME-lox-3xSTOP-lox (JDW 869) | A Gateway compatible middle entry clone containing a loxP flanked SV40 polyA, bGH polyA, and SV40 pA stop cassette |
| 224522 | pME-lox-3xNLS-mCherry-V5-SV40pA-3xstop-lox (JDW 1232) | A Gateway compatible middle entry clone containing loxP flanked 3xNLS-mCherry-V5-3xSTOP cassette |
| 224523 | pME-DreERT2 (JDW 1344) | A Gateway-compatible middle entry clone containing Dre recombinase fused to mutant ERT2 for tamoxifen-inducible recombinase activity |
| 224524 | pME-rox-nls-mCherry-V5-2xStop-rox (JDW 1167) | A Gateway compatible middle entry clone containing a Rox flanked 3xNLS-mCherry-V5-2xStop cassette |
| 224525 | pME EFS-rtTA/rtTA3 (JDW 1230) | A Gateway compatible middle entry clone containing a rtTA-3rd generation transactivator |
| 224526 | p3E-EF1a-pA (JDW 1222) | A Gateway compatible 3' entry clone containing rat EF-1a polyA |
| 224527 | p3E-WPRE-bGH-pA (JDW 1221) | A Gateway compatible 3' entry clone containing a WPRE upstream of Bovine growth hormone polyA to stabilize mRN |
| 224528 | p3E-H2A_mCherry_SV40pA (JDW 967) | A Gateway compatible 3' entry clone containing an H2A mCherry fusion followed by SV40 late polyA |
| 224529 | p3E-V5-mScarlet-I_SV40pA (JDW 968) | A Gateway compatible 3' entry clone containing an V5 mScarlet fusion followed by SV40 late polyA |
| 224530 | p3E-V5-mClover3-pA (JDW 1318) | A Gateway compatible 3' entry clone containing a V5 tagged mClover3 followed by a polyA |
| 224531 | p3E-V5-mTagBFP2 (JDW 871) | A Gateway compatible 3' entry clone containing a V5 tagged mTagBFP2 |
| 224532 | p3E-IRES-H2A-mCherry-SV40-pA (JDW 1256) | A Gateway compatible 3' entry clone containing an IRES H2A mCherry followed by a polyA |
| 224533 | p3E EFS-rtTA/rtTA3 (JDW 1214) | A Gateway compatible 3' entry clone containing the Human EFS promoter driving rtTA-3rd generation transactivator |
| 224534 | p3E-Actin-Vhh-mNeonGreen-HA (JDW 1225) | A Gateway compatible 3' entry clone containing an Actin nanobody fused to mNeonGreen with a c-terminal HA tag |
| 224535 | p3E-IRES-mTagBFP2-pA (JDW 1361) | A Gateway compatible 3' entry clone containing an IRES-FLAG-NLS-mTagBFP2-pA |
| 224536 | p3E-IRES-3xNLS-mScarlet-I3-pA (JDW 1360) | A Gateway compatible 3' entry clone containing IRES-FLAG- NLS-mScarlet-I3 |
| 224537 | p3E-IRES-(n2)StayGold(c4)-pA (JDW 1364) | A Gateway compatible 3' entry clone containing IRES-(n2)StayGold(c4)-pA |
| 224538 | pDONR221-Lifeact- mScarlet-HA (JDW 1247) | A Gateway compatible middle entry clone containing Lifeact-mScarlet-I (far red f-actin reporter) |
| 224539 | pME nls BFP FLAG WPRE bGH pA (JDW 484) | A Gateway compatible middle entry clone containing a 3x-NLS-mTagBFP2-FLAG for labeling the nucleus |
| 224540 | pME nls EGFP WPRE bGH pA (JDW 488) | A Gateway compatible middle entry clone containing a 3x-NLS-EGFP for labeling the nucleus |
| 224543 | pME H2B mCerulean (JDW 1150) | A Gateway compatible middle entry clone containing a histone H2B fusion to mCerulean to label the nucleus |
| 224544 | p5E 5X C120 c-fos min pro (JDW 1241) | A Gateway compatible 5' entry clone containing 5 TAEL transcription factor binding sites and a minimal c-Fos promoter |
| 224545 | p3E-WPRE-SV40-pA (JDW 922) | A Gateway compatible 3' entry clone containing a woodchuck herpes simplex virus regulatory element (WPRE) to stabilize mRNA upstream of an SV40 polyA |
| 229807 | pCAGEN-rtTA-IRES-nls-EGFP-WPRE-bGH-pA (JDW 410) | A CAGGS driven tet-on transactivator with an nls-EGFP reporter |
| 229808 | pB-Puro-DEST (JDW 925) | A puromycin selectable, PiggyBac destination vector compatible with 4-way multi-site gateway cloning system |
| 229809 | pCAGEN-Dre-nls (JDW 1131) | A CAGGS-driven dre recombinase followed by a c-terminal 1x nls sequence |
| 229810 | pTol2-fli1-lsl-AmCyan-mScarlet; hsp70-zfCre-I-BFP (JDW 1234) | A Tol2 based expression vector containing fli1 driven switch reporter (loxP flanked AmCyan followed by a mScarlet-I) and a hsp70-TagBFP-zCre in the opposite direction. |
| 229811 | pTol2-Ubi-lexPR-nls-GFP-pA (JDW 1316) | A Tol2 based expression vector for RU486 inducible expression of GFP in the nucleus (1st generation) |
| 229812 | pCAGEN-nls-mCherry-stop-IRES-myr-EGFP (JDW 1326) | A CAGGS driven expression vector containing an nls-mCherry followed by an IRES and then a myristoylated EGFP for labeling the cell membrane. |
| 229813 | pTol2-Ubi-lexA-oMDC-pA-lexO-cFos-nlsGFP-pA (JDW 1382) | A Tol2 based expression vector for RU486 inducible expression of GFP in the nucleus with a destablized lex transactivator and a c-fos minimal promoter. |
| 229814 | pTol2-Ubi-lsl-EGFP-nls-mCherry-hsp70-zCreI-mTagBFP2 (JDW 1235) | A Tol2 based expression vector containing an Ubi driven loxP flanked GFP followed by an nls-mCherry reporter with hsp70-TagBFP-zCre in the opposite direction. |
| 229815 | pCAGEN-WPRE-bGH-DEST (JDW 1309) | A CAGGS driven, gateway compatible destination vector that contains a WPRE downstream of the destination cassette to stabilize transcripts |
| 229816 | pTol2-Ubi-lsl-EGFP-nls-mCherry:fli1aep-CreI-zf1-BFP (JDW 1236) | A Tol2 based expression vector containing an Ubi driven loxP flanked GFP followed by an nls-mCherry reporter with fli1-TagBFP-zCre in the opposite direction. |
| 229817 | pTol2-DEST-Hsp70-zCreI-mTagBFP2d-2xins (JDW 1002) | A gateway compatible Tol2 destination vector containing the Hsp70 promoter driving Cre and TagBFP followed by 2 cHS4 insulator cores. |
| 229819 | pDestTol2pA-2xIns-fli1ep-zCreI-BFP (JDW 1218) | A gateway compatible Tol2 destination vector containing the fli1a promoter driving Cre and TagBFP in endothelial cells followed by 2 cHS4 insulator cores. |
| 229820 | pAAV-Tet-Off-FLEX-DEST (JDW 1186) | An AAV, tet-off, gateway-compatible destination vector with cre-dependent expression of the insert. EFS driving destablized rTA |
| 229821 | pB-DEST (JDW 1205) | A PiggyBac destination vector compatible with gateway cloning system. |
| 229822 | pGLAST-DEST (JDW 1009) | A single site gateway compatible destination vector with an upstream GLAST promotor to drive expression in glial cells. |
| 229823 | pCAGEN-Luciferase-P2A-H2A-mCherry (JDW 1129) | A CAGGS driven luciferase reporter followed by a P2A cleavage peptide and an H2A mCherry cassette for nuclear labeling. |
| 229824 | pB-Tet-Off_FLEX_Luc-P2A-H2A-mCherry (JDW 970) | A PiggyBac vector with a cre-dependent dual luciferase / nuclear mCherry reporter. In the presence of cre, this tet-off vector will be ubiquitously expressed in the absence of dox/tet |
| 229825 | pB-Neo-DEST (JDW 940) | A neo selectable, PiggyBac destination vector compatible with gateway cloning system. |
| 229826 | pAAV-Tet-Off-FLEX-Lifeact-mScarlet-HA-WPRE-pA (JDW 721) | Tet-off, AAV vector for cre dependent expression of Lifeact mScarlet tagged with HA with EFS (minimal EF1a promoter or EF1a short/ EFS) driving destablized tTA (d2tTA) |
| 229827 | pCAGEN nls-BFP-FLAG-WPRE-bgh-pA (JDW 452) | A CAGGS driven nls BFP with a FLAG tag followed by a WPRE-bGH polyA for stable transcripts |
| 229828 | pB_Tet-Off_Flex_DEST_EFS_mODC_tTA (JDW 936) | A Piggybac-compatible, cre-dependent, Tet-off destination vector with a downstream WPRE. EFS driving destabilized tTA |
| 229829 | p5E-MCS-c-Fos-b-globin (JDW 1237) | A gateway compatible 5' entry clone containing an MCS upstream of the murine c-fos minimal promotor and b-globin intron. |
| 229830 | p5E-Crestin-pro (JDW 1320) | A gateway compatible 5' entry clone containing the crestin gene followed by the mus musculus c Fos minimal promoter and then a rabbit beta globin intron to drive neural crest-specific gene expression |
| 229831 | pTol2-DEST-hsp70-zCreI-BFP (JDW 1184) | A gateway compatible Tol2 destination vector containing the hsp70 promoter driving Cre and TagBFP in endothelial cells followed by 2 cHS4 insulator cores. |
| 229832 | pME-MCS (JDW 455) | A gateway compatible middle entry clone containing an MCS (mutiple cloning site). |
| 229833 | pXMLC2-DEST (JDW 7) | A gateway compatible destination vector containing the Xenopous myosin light chain 2 promoter for embryonic and adult cardiomyocyte expression. |
| 229834 | pCAGEN-DV-IRES-myr-BFP (JDW 491) | A CAGGS driven gateway compatible destination vector that contains an IRES-myristoylated-TagBFP for labeling the cell membrane |
| 229835 | pBeta-MHC-promoter DV (JDW 26) | A gateway compatible destination vector containing the murine beta-MHC promoter for embryonic and postnatal cardiomyocyte expression. |
| 229836 | p5E-dr-myl7-pro (JDW 1148) | A gateway compatible 5' entry clone containing the mysoin light chain seven (myl7/cmlc2) promoter for cardiomyocyte specific expression. |
| 229837 | pB-Tet-On-Luciferase-P2A-H2A-mCherry (JDW 945) | A Piggybac compatible, tet-on, NEO selectable, vector for expressing luciferase followed by a P2A cleavage peptide and H2A fused mCherry. |
| 229838 | pTol2-Ubi-LexA-oMDC-pA-LexOp-nlsGFP-pA (JDW 1381) | A Tol2 based expression vector for RU486 inducible expression of GFP in the nucleus with a destablized lex transactivator. |
| 229839 | pB-TetOn-DEST-hEF1a-mODC-rtTA-IRES-NEO (JDW 1130) | A CAGGS driven, piggybac compatible, tet-on expression vector containing a luciferase reporter followed by a P2A cleavage peptide and H2A fused mCherry for nuclear labeling. |
| 229840 | pB-Tet-On-Luciferase-P2A-H2A-mCherry (JDW 1154) | A Piggybac compatible, tet-on, dual reporter encoding luciferase followed by a P2A cleavage peptide and then an H2A fused mCherry for nuclear labeling. |
| 229841 | pTol2-Hsp70-H2B-mCerulean-lsl-mScarlet-fli1-zCre-I-BFP (JDW 1233) | A Tol2 based expression vector with the Hsp70 promoter driving a cre dependent switch reporter. Contains an endothelial driven cre |
| 229842 | pCS2-phiC31o (JDW 453) | A CMV-driven Phi31 recombinase. |
| 229843 | p5E-Dll4-F2-E1B-B-Globin (JDW 1366) | A gateway compatible 5' entry clone containing the murine Dll4 F2 arterial specific enhancer upstream of a minimal E1b promoter and intron for arterial and endocardial expression. |
| 229844 | pME-StayGoldc4-Giantin (JDW 1355) | A gateway compatible middle entry clone containing a V5 tagged n2StayGoldc4 flourescent protein |
| 229845 | pCAGEN-Vhh-LaminB-mNeonGreen-IRES-myr-BFP (JDW 1353) | A CAGGS-driven expression vector containing a vhh-Lamin nanobody for nuclear lamina labeling followed by an IRES and a myristoylated TagBFP for labeling the cell membrane. |