Difference between revisions of "Part:BBa K5034201"

(→‎Experimental Characterization and results)
(→‎Sequence and Features)
Line 9: Line 9:
  
 
===Sequence and Features===
 
===Sequence and Features===
 +
This backbone consists of several parts, including promoters, terminators, resistance genes, and prefix and suffix we added.
 +
 +
<html>
 +
<div align="center">
 +
<img style="width:50%;height:auto;" src="https://static.igem.wiki/teams/5034/engineering/pbbr1mcs-terminator.png">
 +
<p>
 +
Figure 1: Plasmid diagram of backbone
 +
</p>
 +
</div>
 +
</html>
 +
 
<partinfo>BBa_K5034201 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K5034201 SequenceAndFeatures</partinfo>
  

Revision as of 07:22, 2 October 2024


pBBR1MCS-terminator plasmid

Description

The pBBR1MCS plasmid is a synthetic plasmid backbone used in various genetic engineering applications in microbial systems. We modified the original pBBR1MCS and added a double terminator rrnBT1-T7Te(BBa_B0015) downstream the Biobrick suffix.

Sequence and Features

This backbone consists of several parts, including promoters, terminators, resistance genes, and prefix and suffix we added.

Figure 1: Plasmid diagram of backbone


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 4981
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 2834
    Illegal NotI site found at 4987
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 4981
    Illegal BglII site found at 3580
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 4981
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 4981
    Plasmid lacks a suffix.
    Illegal XbaI site found at 4996
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NgoMIV site found at 562
    Illegal NgoMIV site found at 4244
    Illegal NgoMIV site found at 4527
    Illegal AgeI site found at 402
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.

Experimental Characterization and results

We performed double enzyme digestion on the plasmid and obtained a fragment of approximately 5000bp(Fig.1)

Figure 1: Fragment obtained by double enzyme digestion of plasmid backbone

Applications

1. It can be used in microbial fuel cells to study and enhance the extracellular electron transfer capabilities such as in S. oneidensis.

2. It can be used as a backbone for CRISPR-based genome editing systems, enabling precise genetic modifications.

Reference

1. Kovach ME, Elzer PH, Hill DS, Robertson GT, Farris MA, Roop RM 2nd, Peterson KM. Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes. Gene. 1995 Dec 1;166(1):175-6. doi: 10.1016/0378-1119(95)00584-1. PMID: 8529885.