Difference between revisions of "Part:BBa K1898450:Design"

 
 
Line 7: Line 7:
  
 
===Design Notes===
 
===Design Notes===
−
We made sure that there was no Ecor1, Xbal, Spel1, and Pst1 cutting sites in the DNA. Primers were designed to remove the stop codon from cryab and were synthesized by Tri-I. Our assembly method BioBrick RFC[10] shifted the open reading frame and created a stop codon in between cryab and 10x histidine tag. The sequence was sent to Missiontech to add two base pairs in between the two genes.
 
  
 +
We removed the stop codon in CRYAB when designing the construct.
  
 +
The following primers are designed and used to move the DNA synthesized by IDT into iGEM Biobrick:
  
 +
forward: 5' ATATgAATTCgCggCCg 3' (17)
  
−
===Source===
+
reverse: 5' ATATCTgCAgCggCC 3' (15)
  
−
the cDNA of cryab was ordered from OriGene. The promoter and rbs was from iGEM distribution kit (BBa_K880005) and His-Term was from an intermediate part we constructed (BBa_K1898500).  
+
Our assembly method BioBrick RFC[10] shifted the open reading frame and created a stop codon in between CRYAB and 10x histidine tag. This is showed in this part's main page under sequencing. The sequence was sent to mutagenesis to add two base pairs in between the two genes.
 +
 
 +
 
 +
===Source===
  
 +
CRYAB was synthesized by IDT
  
−
===References===
+
Primers were ordered from Tri-I Biotech and mutagenesis was done by MissionBiotech.

Latest revision as of 03:56, 19 October 2016


Strong promoter + Strong RBS + CRYAB + 10x Histidine tag + Double terminator


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
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 87
    Illegal BamHI site found at 429
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 514
    Illegal SapI.rc site found at 374


Design Notes

We removed the stop codon in CRYAB when designing the construct.

The following primers are designed and used to move the DNA synthesized by IDT into iGEM Biobrick:

forward: 5' ATATgAATTCgCggCCg 3' (17)

reverse: 5' ATATCTgCAgCggCC 3' (15)

Our assembly method BioBrick RFC[10] shifted the open reading frame and created a stop codon in between CRYAB and 10x histidine tag. This is showed in this part's main page under sequencing. The sequence was sent to mutagenesis to add two base pairs in between the two genes.


Source

CRYAB was synthesized by IDT

Primers were ordered from Tri-I Biotech and mutagenesis was done by MissionBiotech.