Subcloning Tasks
| ID | Name | DNA Size | Vector | Parent | Method |
Date | Enzyme |
Expression | Note | |
| 0 | 3161 |
pET28b-GST--Myc-HA-Flag-V5 |
224 | 433 pDB.His.GST |
3159 pMiniT2-1FRG-HA-VL-kappa |
Nde1-Xho1 | ||||
| 3127 |
pET28b-qBeta-qBeta-6XHis |
PCR qbeta and clone back into 3097 | Nde1-HindIII | C-His tag | PCR product is in pT&A3, 997, sequenced. | |||||
| 812 |
pET30b-phoA |
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| 3074 |
pET30b-HA-avitag-BirA |
in pJet1.2 is good, subclone into pET28b, no his tag, need to subclone into pT7T30-Avi-BirA (still no his tag) | ||||||||
| 938 | PNGase F, | 1145 | pET30b |
Nde1-Xho1 | pJet1.2, sequencing is good, need to subclone into pET30b |
C-His | ||||
| 939 | Tte-Exo III | 780 | pET30b |
Nde1-Xho1 |
pJet1.2 sequencing is good, need to subclone into pET30b |
C-His | ||||
| 942 | VVN endonuclease | 716 | pET28b-pMBP |
Nco1-Xho1 |
subclone from 942-3-PJET1-2 |
C-His | ||||
| 3122 | Crm197, secreted | 1743 | pET30b |
Nde1-Xho1 |
pJet1.2 |
C-His | ||||
| 931 | PAK16D ligase | 2267 | pET28b | Nde1-HindIII-Xho1 |
pJet1.2 fragment 2 is good fragment 1 is also good |
N-His | ||||
| 945 |
pT7A30-pMBP |
pT7A3 | Xba1-Nco1 |
PCR from 1281 pMAL-p5E or 1282 pMAL-p5E-AvPAL | result | |||||
| 947 |
pT7A30-6His-pMBP |
result | ||||||||
| 949 |
pET28b-GST-sfGFP-Myc-HA-Flag-V5 |
433 546 |
||||||||
| 824 |
pET28b-VVTopo |
|||||||||
| 985 |
pCold-GST-3C-AP205-M13-g3p |
2047 pCold-C6H |
983 pMiniT2-973-2 |
insert: Nde1-Sal1, vector: Nde1-Xho1 |
Result | |||||
| 984 |
pT7A3-6H-SBP-SUMO-3C |
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| 986 |
pT7A3-6His-NLS-Cas9-NLS-6xHis |
need to sequencing, expression screening | result |
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