Difference between revisions of "Part:BBa K2259093"
(→‎Usage with SynORI (Framework for multi-plasmid systems))
Line 30: Line 30:
 
SynORI is a framework for multi-plasmid systems created by ''Vilnius-Lithuania 2017'' which enables quick and easy workflow with multiple plasmids, while also allowing to freely pick and modulate copy number for every unique plasmid group! Read more about [http://2017.igem.org/Team:Vilnius-Lithuania SynORI here]!
 
SynORI is a framework for multi-plasmid systems created by ''Vilnius-Lithuania 2017'' which enables quick and easy workflow with multiple plasmids, while also allowing to freely pick and modulate copy number for every unique plasmid group! Read more about [http://2017.igem.org/Team:Vilnius-Lithuania SynORI here]!
  
−
===Regulative RNA II molecule in SynORI===
+
===This part in SynORI===
−
RNA II gene is foundational and central biobrick of SynORI system, and by far the only one that is mandatory for framework to run.
+
Engineering an improved, functional base vector 2.0 was crucial for SynORI framework, because building a synthetic origin of replication required an empty biobrick site and no origin of replication in backbone. One can then replace the pUC origin of replication to SynORI system parts. Once the modular SynORI system is built, it can be transfered to another plasmid location and biobricks are then free to use for other projects required.  
−
The two main functions of RNA II is as follows:
+
−
# Initiating plasmid replication
+
−
# Interacting with RNA I of specific plasmid group [[#Specific RNA II versions in multi-plasmid systems|(See below)]]
+
−
 
+
−
 
+
−
=== RNA II and RNA I in the engineering of unique plasmid  groups for multi-plasmid system===
+
−
 
+
−
RNA II molecule interacts with inhibitory RNA I molecule with three secondary structure RNA stem loops. In order to create plasmid groups with independent copy number control, one group's RNA II molecule must interact only with the same group's RNA I molecule.
+
−
 
+
−
<b>For example</b> if there are two plasmid groups in a cell - A and B - RNA II of A group
+
−
would only interact with RNA I A, and not RNA I B.
+
−
 
+
−
[[Image:RnainteractionIII.png|center|500px|thumb|<b>Figure 1. </b> RNA I AND II group interaction example]]
+
−
 
+
−
See the [https://parts.igem.org/Part:BBa_K2259000:Design Design] section or [http://2017.igem.org/Team:Vilnius-Lithuania Vilnius-Lithuania 2017 team wiki] for more insight about our synthetic origin of replication (SynORI).
+
  
 +
See the [http://2017.igem.org/Team:Vilnius-Lithuania Vilnius-Lithuania 2017 team wiki] for more insight about our synthetic origin of replication (SynORI).
  
 
=Characterization of RNA II (Vilnius-Lithuania 2017)=
 
=Characterization of RNA II (Vilnius-Lithuania 2017)=

Revision as of 08:59, 31 October 2017


SynORI base vector 2.0 insert

This composite part is designed to replace the base vector's Part:BBa_I51020 insert which is no longer available because of the ccdB part containing construct withdrawal.

Vector backbone which does not contain this insert can be [found here.]

These combined sequences result in the new Base vector 2.0, which instead of ccdB poison uses blue white screening to select for correct colonies when building a custom plasmid.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 462
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]



Introduction

Usage with SynORI (Framework for multi-plasmid systems)

About SynORI

Aboutsynoritry1.png

SynORI is a framework for multi-plasmid systems created by Vilnius-Lithuania 2017 which enables quick and easy workflow with multiple plasmids, while also allowing to freely pick and modulate copy number for every unique plasmid group! Read more about [http://2017.igem.org/Team:Vilnius-Lithuania SynORI here]!

This part in SynORI

Engineering an improved, functional base vector 2.0 was crucial for SynORI framework, because building a synthetic origin of replication required an empty biobrick site and no origin of replication in backbone. One can then replace the pUC origin of replication to SynORI system parts. Once the modular SynORI system is built, it can be transfered to another plasmid location and biobricks are then free to use for other projects required.

See the [http://2017.igem.org/Team:Vilnius-Lithuania Vilnius-Lithuania 2017 team wiki] for more insight about our synthetic origin of replication (SynORI).

Characterization of RNA II (Vilnius-Lithuania 2017)

RNA I inactivation in wild type replicon

References