View source for Part:pSB1C3

__NOTOC__
<partinfo>pSB1C3 short</partinfo>

pSB1C3 is a high copy number plasmid (RFC [10]) carrying chloramphenicol resistance.

The replication origin is a pUC19-derived pMB1 (copy number of 100-300 per cell).

pSB1C3 has terminators bracketing its MCS which are designed to prevent transcription from *inside* the MCS from reading out into the vector. The efficiency of these terminators is known to be < 100%. Ideally we would construct a future set of terminators for bracketing a MCS that were 100% efficient in terminating both into and out of the MCS region.

'''pSB1C3 is the designated Registry shipping plasmid backbone. All submissions for iGEM competitions must be submitted using pSB1C3.'''

Go to [https://parts.igem.org/DNA_Submission_Instructions this page] for more information on how to ship submissions to iGEM HQ.



===Sample Location===
The distribution kits contain many samples of pSB1C3 as it is the Registry's main backbone for shipping and maintaining samples. Our recommendation when cloning a part into pSB1C3 is that you can use either linearized plasmid backbone provided in the kits, or pSB1C3 with a reporter (BBa_J04450).

'''2017 Kit Plate 4 - Well 4B''' has a sample of BBa_J04450 in pSB1C3.



===Usage and Biology===

In Spring 2011, pSB1C3 was sequenced using several primers: Pre-R, Suff-F, VF2, VR, and internal primers for the origin and resistance. The reference sequence below matches the most recent sequencing verification of pSB1C3 (w/ BBa_J04450, located in SP 4000 Well 2A).

[http://2017.igem.org/Team:USP-Brazil Team USP-Brazil 2017] improved the function of part by showing that it can be used to sucessfully transform another chassis from the Enterobacteriaceae family, <i> Pantoea agglomerans </i><br>
(<small>--[[User:lubdub|lubdub]] 02:38, 30 October 2017 (UTC)</small>) 

===Improvement by Evry_Paris-Saclay 2017===
The [http://2017.igem.org/Team:Evry_Paris-Saclay Evry_Paris-Saclay 2017 team] made two improvements of the pSB1C3 backbone [[Part:BBa_K2448038|BBa_K2448038]] and [[Part:BBa_K2448036|BBa_K2448036]].
*[[Part:BBa_K2448038|BBa_K2448038]]: pSB1C3 is high copy number plasmid and the iGEM workhorse for gene expression. However, when it comes to the of use LacI regulated promoters, very frequently significant leakage is observe due to its important copy number, which leads to important regulation problems. This issue can be addressed either by including the LacI coding sequence into the construct or by using only LacI overexpressing cells. To facilitate further use of pSB1C3, we designed a pSB1C3 with a built-in LacI coding sequence under the control of a mutated version of its own natural promoter known as LacIq which leads to a 10-fold increase in LacI expression compared to the natural promoter. This improved backbone allows a fine regulation of LacI related parts.
*[[Part:BBa_K2448036|BBa_K2448036]]: the pSB1C3 backbone vector contains a BsmBI cloning site within the chloramphenicol resistance gene. Its presence prevents from using the Golden Gate assembly technique with this backbone. To circumvent this issue, we performed a site-directed mutagenesis (G1385C) and created the pSB1C3 BsaI free backbone.

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<span class='h3bb'>Sequence and Features</span>
<partinfo>pSB1C3 SequenceAndFeatures</partinfo>


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===Functional Parameters===
<partinfo>pSB1C3 parameters</partinfo>
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