Difference between revisions of "Part:pSB1A3:Experience"

Latest revision as of 08:26, 29 April 2012

This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of pSB1A3

Characterization

User Reviews

UNIQfc3633cffbfcdfdb-partinfo-00000000-QINU

•••••

Antiquity

This review comes from the old result system and indicates that this part worked in some test.

•••

Kahaynes

Our team ([http://2006.igem.org/Davidson_2006 Davidson College]) has used this vector in E. coli strain JM109 extensively. Clones are Amp resistant and are recovered in reasonable yields from mini preps. One drawback is that the vector has read-through transcription that comes into the cloned BioBrick part from both directions. This problem is a major one when an "off state" is critical for the control of a device. Parts without a promoter are expressed in this vector, regardless of orientation (see pSB1A7 Part Design for details). Reverse read-through might be from the Amp resistance gene. Forward read-through might be from a cryptic promoter, since no known genes in the backbone are in the plus orientation.

--Kahaynes 15:48, 23 October 2006 (EDT)

•

siegeljb

"***WARNING***" The RFP version of this part is actually in pSB1A2 and NOT pSB1A3. This finding has been personally sequenced confirmed in addition to being confirmed in the 2009 QC (look at the VR sequence of the RFP version of this part from the 2009 distribution: QC09_P667_W41590_VR.ab1). This means that the built in terminator is NOT there. The ccdB version appears to actually be pSB1A3 which does contain the terminator. Look [1]here.

;

•

Marc.mtk

"***WARNING***" [http://2009.igem.org/Team:SDU-Denmark We] can confirm the findings of siegeljb, The contents of well 1K in this years distribution plates(supposedly BBa_J04450 in pSB1A3) is in fact BBa_J04450 in pSB1A2.!!! I did a write up of the data supporting this conclusion [http://2009.igem.org/Team:SDU-Denmark/Parts/backbone_troubles here].

If you still want to use this backbone, get it with p1010 (ccdB) insert instead, that looks like the real deal :)

;

•••

UWash_iGEM2011

Our team utilized this vector to test its Gibson efficiency compared to pGA1C3.Using 1 ng of this vector with 1 ng of our desired insert, we experienced low efficiency among six plates. The average Gibson Assembly efficiency was calculated be be ~11%. This was therefore not used, as there are better yielding vectors now available. For more information about the efficiency of pSB and a comparison of pSB vs pGA vectors, please click [http://2011.igem.org/Team:Washington/Magnetosomes/GibsonResults here!]

UNIQfc3633cffbfcdfdb-partinfo-00000008-QINU

@@ Line 1: / Line 1: @@
 __NOTOC__
 This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
@@ Line 5: / Line 4: @@
 ===Applications of pSB1A3===
+===Characterization===
 ===User Reviews===
@@ Line 33: / Line 34: @@
 <I>Kahaynes</I>
 |width='60%' valign='top'|
-Our team ([http://parts.mit.edu/wiki/index.php/Davidson_2006 Davidson College]) has used this vector in ''E. coli'' strain JM109 extensively. Clones are Amp resistant and are recovered in reasonable yields from mini preps. One drawback is that the vector has read-through transcription that comes into the cloned BioBrick part from both directions. This problem is a major one when an "off state" is critical for the control of a device. Parts without a promoter are expressed in this vector, regardless of orientation (see [https://parts.igem.org/wiki/index.php/Part:BBa_J31009:Design pSB1A4 Part Design] for details). Reverse read-through might be from the Amp resistance gene. Forward read-through might be from a cryptic promoter, since no known genes in the backbone are in the minus (complimentary) orientation.
+Our team ([http://2006.igem.org/Davidson_2006 Davidson College]) has used this vector in ''E. coli'' strain JM109 extensively. Clones are Amp resistant and are recovered in reasonable yields from mini preps. One drawback is that the vector has read-through transcription that comes into the cloned BioBrick part from both directions. This problem is a major one when an "off state" is critical for the control of a device. Parts without a promoter are expressed in this vector, regardless of orientation (see [https://parts.igem.org/wiki/index.php/Part:BBa_J31009:Design pSB1A7 Part Design] for details). Reverse read-through might be from the Amp resistance gene. Forward read-through might be from a cryptic promoter, since no known genes in the backbone are in the plus orientation.
+--[[User:Kahaynes|Kahaynes]] 15:48, 23 October 2006 (EDT)
 |}
+{|width='80%' style='border:1px solid gray'
+|-
+|width='10%'|
+<partinfo>pSB1A3 AddReview 1</partinfo>
+<I>siegeljb</I>
+|width='60%' valign='top'|
+"***WARNING***" The RFP version of this part is actually in pSB1A2 and NOT pSB1A3.  This finding has been personally sequenced confirmed in addition to being confirmed in the 2009 QC (look at the VR sequence of the RFP version of this part from the 2009 distribution: [https://parts.igem.org/cgi/sequencing/get_electrophorogram.cgi?id=7377 QC09_P667_W41590_VR.ab1]).  This means that the built in terminator is NOT there.  The ccdB version appears to actually be pSB1A3 which does contain the terminator.
+Look [https://parts.igem.org/cgi/sequencing/one_blast.cgi?id=3915 ]here.
+|};
+{|width='80%' style='border:1px solid gray'
+|-
+|width='10%'|
+<partinfo>pSB1A3 AddReview 1</partinfo>
+<I>Marc.mtk</I>
+|width='60%' valign='top'|
+"***WARNING***" [http://2009.igem.org/Team:SDU-Denmark We] can confirm the findings of siegeljb, The contents of well 1K in this years distribution plates(supposedly BBa_J04450 in pSB1A3) is in fact <partinfo>BBa_J04450</partinfo> in <partinfo>pSB1A2</partinfo>.!!! I did a write up of the data supporting this conclusion [http://2009.igem.org/Team:SDU-Denmark/Parts/backbone_troubles here].
+If you still want to use this backbone, get it with p1010 (ccdB) insert instead, that looks like the real deal :)
+|};
+{|width='80%' style='border:1px solid gray'
+|-
+|width='10%'|
+<partinfo>pSB1A3 AddReview 3</partinfo>
+<I>UWash_iGEM2011</I>
+|width='60%' valign='top'|
+Our team utilized this vector to test its Gibson efficiency compared to pGA1C3.Using 1 ng of this vector with 1 ng of our desired insert, we experienced low efficiency among six plates. The average Gibson Assembly efficiency was calculated be be ~11%. This was therefore not used, as there are better yielding vectors now available.
+For more information about the efficiency of pSB and a comparison of pSB  vs pGA vectors, please click [http://2011.igem.org/Team:Washington/Magnetosomes/GibsonResults here!]
 <!-- DON'T DELETE --><partinfo>pSB1A3 EndReviews</partinfo>