Difference between revisions of "Part:BBa K3866038"
 
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===Usage and Biology===
 
===Usage and Biology===
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This TU includes the <i>sbm</i> gene placed under the control of the arabinosed-induced promoter <bbpart>BBa_K3866000</bbpart>. Initially, the level 1 cloning was performed using the trc promoter <bbpart>BBa_K3866001</bbpart>, though we weren't able to isolate a colony with the desired construct, as there is a possibility that the bacteria's metabolism went out of balance so no colonies survived to be chosen for screening. Instead we used the arabinose-indused promoter, which seemed to be working as expected.
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This TU includes the <i>sfGFP</i> gene placed under the control of the constitutive Anderson promoter J23115 <bbpart>BBa_J23115</bbpart>.
  
 
===Design Notes===
 
===Design Notes===
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The coding sequence was domesticated. We removed BsmBI and BsaI sites in order to be compatible with GoldenBraid and MoClo. The sequence is cloned in a PDGB3_α1 vector and has overhangs compatible for GoldenBraid cloning.
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The coding sequence was domesticated. We removed BsmBI and BsaI sites in order to be compatible with GoldenBraid and MoClo.
  
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[[Image:T--Thessaly--J23115-sfGFP-term.png|800px|thumb|none|<I><b>Figure 1.</b> The first level α module of the Propionate Production: α1:ParaBAD:RBS-smb-Double terminator </i>]]
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[[Image:T--Thessaly--J23115-sfGFP-term.png|800px|thumb|none|<I><b>Figure 1.</b> The level α module of the sfGFP Translational Unit: α1: J23115:RBS-sfGFP-Double terminator</i>]]
  
 
===Verification of Cloning===
 
===Verification of Cloning===
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[[File:T--Thessaly--gfpgel.png|800px|thumb|none|<i><b>Figure 2.</b>: (C=Cut, U=Uncut) Restriction digestion of α1-sbm with: EcoRI, Expected bands: 6345bp, 2947bp, 597bp, Positive result: C2</i>]]
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[[File:T--Thessaly--gfpgel.png|800px|thumb|none|<i><b>Figure 2.</b>: (C=Cut, U=Uncut) Restriction digestion of α-sfGFP TU (samples 3,4,5) with: HindIII, Expected bands: 2847bp, 801bp, 146bp, Positive result: C5</i>]]
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===Experimental Use and Experience===
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This part showed functionality at the following parts: <bbpart>BBa_K3866029</bbpart>, <bbpart>BBa_K3866031</bbpart>
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===Functional Parameters===
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<partinfo>BBa_K3866038 parameters</partinfo>
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===Sequence and Features===
 
===Sequence and Features===
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<partinfo>BBa_K3866026 SequenceAndFeatures</partinfo>
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<partinfo>BBa_K3866038 SequenceAndFeatures</partinfo>

Latest revision as of 12:13, 17 October 2021


J23115:sfGFP:Terminator

Usage and Biology

This TU includes the sfGFP gene placed under the control of the constitutive Anderson promoter J23115 BBa_J23115.

Design Notes

The coding sequence was domesticated. We removed BsmBI and BsaI sites in order to be compatible with GoldenBraid and MoClo.

Figure 1. The level α module of the sfGFP Translational Unit: α1: J23115:RBS-sfGFP-Double terminator

Verification of Cloning

Figure 2.: (C=Cut, U=Uncut) Restriction digestion of α-sfGFP TU (samples 3,4,5) with: HindIII, Expected bands: 2847bp, 801bp, 146bp, Positive result: C5

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 192
  • 1000
    COMPATIBLE WITH RFC[1000]