Difference between revisions of "Part:pSB4A3"
Smelissali (Talk | contribs) |
Smelissali (Talk | contribs) |
||
| Line 2: | Line 2: | ||
<partinfo>pSB4A3 short</partinfo> | <partinfo>pSB4A3 short</partinfo> | ||
| − | pSB4A3-1 is a low copy plasmid with ampicillin resistance. It has a repA pSC101-derived replication origin. (Copy number ~10-12, Lutz and Bujard, 1997.) | + | pSB4A3-1 is a low copy plasmid with ampicillin resistance. It has a repA pSC101-derived replication origin, and thus can be co-expressed in the same host cell as Biobrick plasmids with the sufficiently dissimilar pMB1 origin (see [http://openwetware.org/wiki/Vectors#Escherichia%20coli OpenWetware's documentation for more information]). (Copy number ~10-12, Lutz and Bujard, 1997.) |
| + | |||
pSB4A3 has a terminator upstream of its MCS, which is oriented to prevent transcription from *inside* the MCS from reading out into any DNA outside the MCS.The orientation of the pBLA promoter driving the AmpR gene is such that relatively little transcription should be coming into the MCS from upstream. A second terminator (E.coli His operon-derived) is downstream of the MCS, again insulating the vector from transcription reading out of the MCS. Ideally, future versions of standard biobrick vectors would have terminators bracketing the MCS that were 100% efficient in terminating transcription both into and out of the MCS region. | pSB4A3 has a terminator upstream of its MCS, which is oriented to prevent transcription from *inside* the MCS from reading out into any DNA outside the MCS.The orientation of the pBLA promoter driving the AmpR gene is such that relatively little transcription should be coming into the MCS from upstream. A second terminator (E.coli His operon-derived) is downstream of the MCS, again insulating the vector from transcription reading out of the MCS. Ideally, future versions of standard biobrick vectors would have terminators bracketing the MCS that were 100% efficient in terminating transcription both into and out of the MCS region. | ||
Revision as of 21:45, 12 June 2007
pSB4A3 (Replaced by pSB4A5)
pSB4A3-1 is a low copy plasmid with ampicillin resistance. It has a repA pSC101-derived replication origin, and thus can be co-expressed in the same host cell as Biobrick plasmids with the sufficiently dissimilar pMB1 origin (see [http://openwetware.org/wiki/Vectors#Escherichia%20coli OpenWetware's documentation for more information]). (Copy number ~10-12, Lutz and Bujard, 1997.)
pSB4A3 has a terminator upstream of its MCS, which is oriented to prevent transcription from *inside* the MCS from reading out into any DNA outside the MCS.The orientation of the pBLA promoter driving the AmpR gene is such that relatively little transcription should be coming into the MCS from upstream. A second terminator (E.coli His operon-derived) is downstream of the MCS, again insulating the vector from transcription reading out of the MCS. Ideally, future versions of standard biobrick vectors would have terminators bracketing the MCS that were 100% efficient in terminating transcription both into and out of the MCS region.
Usage and Biology
- Primers for these Biobrick vectors can be found in Part:BBa_G00100 (aka VF2) and Part:BBa_G00101 (aka. VR)
- Annotated [http://www.biology.utah.edu/jorgensen/wayned/ape/ APE] file for pSB4A3
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 3318
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NotI site found at 9
Illegal NotI site found at 3324 - 21INCOMPATIBLE WITH RFC[21]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 3318
Illegal XhoI site found at 177 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found at 3318
Illegal suffix found at 2 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found at 3318
Plasmid lacks a suffix.
Illegal XbaI site found at 3333
Illegal SpeI site found at 2
Illegal PstI site found at 16 - 1000INCOMPATIBLE WITH RFC[1000]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal BsaI.rc site found at 2360