PSB1A7
From 2006.igem.org
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[[Image:pSB1A7.gif|600px]] | [[Image:pSB1A7.gif|600px]] | ||
| - | To prevent uncontrolled expression of parts within our device, we built insulated cloning vector '''pSB1A7''' (<partinfo>BBa_J31009</partinfo>). pSB1A7 was built using the pre-existing <partinfo>pSB1A3</partinfo> vector. The terminators in pSB1A3 are designed to prevent read-through from inside the device out into the | + | To prevent uncontrolled expression of parts within our device, we built insulated cloning vector '''pSB1A7''' (<partinfo>BBa_J31009</partinfo>). pSB1A7 was built using the pre-existing <partinfo>pSB1A3</partinfo> vector. The terminators in pSB1A3 are designed to prevent read-through from inside the device out into the vector (purple arrows) but the vector is not designed to stop read-through into the device (red arrows). We added double terminators (<partinfo>BBa_B0015</partinfo> shown as solid red and hatched hexagons) in opposite orientations at each end of the BioBrick cloning site region to shield the insert from transcription read-through. When the pancakes are cloned into pSB1A7, tetracycline resistance is dependent on the proper orientation of the two DNA pancakes. |
See [http://partsregistry.org/Part:BBa_J31009:Design the part design page] for details on how pSB1A7 was built. | See [http://partsregistry.org/Part:BBa_J31009:Design the part design page] for details on how pSB1A7 was built. | ||
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| + | ===pSB1A7 is not compatible with parts carrying B0015=== | ||
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| + | Our new cloning vector pSB1A7 insulates our parts from read-through transcription, but this vector does not accept parts that carry the B0015 double terminator. Our flipping device was originally designed with a B0015 near the end. Several attempts to clone parts carrying B0015 into pSB1A7 failed. This problem prompted a [[PSB1A7 Cloning Solution|redesign of our system]]. | ||
Latest revision as of 19:05, 31 October 2006
Our new pSB1A7 cloning vector blocks read-through transcription
To prevent uncontrolled expression of parts within our device, we built insulated cloning vector pSB1A7 (). pSB1A7 was built using the pre-existing vector. The terminators in pSB1A3 are designed to prevent read-through from inside the device out into the vector (purple arrows) but the vector is not designed to stop read-through into the device (red arrows). We added double terminators ( shown as solid red and hatched hexagons) in opposite orientations at each end of the BioBrick cloning site region to shield the insert from transcription read-through. When the pancakes are cloned into pSB1A7, tetracycline resistance is dependent on the proper orientation of the two DNA pancakes.
See [http://partsregistry.org/Part:BBa_J31009:Design the part design page] for details on how pSB1A7 was built.
pSB1A7 is not compatible with parts carrying B0015
Our new cloning vector pSB1A7 insulates our parts from read-through transcription, but this vector does not accept parts that carry the B0015 double terminator. Our flipping device was originally designed with a B0015 near the end. Several attempts to clone parts carrying B0015 into pSB1A7 failed. This problem prompted a redesign of our system.
