Difference between revisions of "Part:BBa J31006:Design"

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===Design Notes===
 
===Design Notes===
  
It was cloned into PsB1A2 plasmids.
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BBa J31006 was cloned into vector pSB1A2. The Biobricks on this part are not wild type, but the cut sites are still viable.  
 
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===Biobricks===
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The Biobricks on this part are not wild type, but the cut sites are still viable.  
+
  
 
{| width="800px" cellspacing="5"
 
{| width="800px" cellspacing="5"
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|- valign="top"
 
|- valign="top"
 
| style="width:180px" | '''BBa_J31001 Cloning Sites'''  
 
| style="width:180px" | '''BBa_J31001 Cloning Sites'''  
| style="background:lightgrey" |<font face="courier">5'--GAATTC GCGGCCGC T TCTAGA <font color='blue'>*</font> --Hin coding-- <font color='purple'>*</font> ACTAGT <font color='darkgreen'>T</font> GCGGCCG<font color='magenta'>C</font>CTGCAG--<br>3'--CTTAAG CGCCGGCG <font color='red'>*</font> AGATCT <font color='blue'>*</font> -------------- <font color='purple'>*</font> TGATCA <font color='darkgreen'>A</font> CGCCGGC<font color='magenta'>G</font>GACGTC--</font><br><br>
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| style="background:lightgrey" |<font face="courier">5'--GAATTC GCGGCCGC T TCTAGA <font color='blue'>*</font> --Tet coding-- <font color='purple'>*</font> ACTAGT A GCGGCCG CTGCAG--<br>3'--CTTAAG CGCCGGCG A AGATCT <font color='blue'>*</font> -------------- <font color='purple'>*</font> TGATCA T CGCCGGC GACGTC--</font><br><br>
 
'''Prefix'''<br>There is <font color='blue'>no G spacer (*)</font> between the XbaI and the Hin coding region.<br>
 
'''Prefix'''<br>There is <font color='blue'>no G spacer (*)</font> between the XbaI and the Hin coding region.<br>
'''Suffix'''<br>There is <font color='purple'>no T spacer (*)</font> between the Hin coding region and the SpeI site.<br>The A spacer between the SpeI and the NotI has changed to a <font color='darkgreen'>T</font>.<br>There is an extra <font color='magenta'>C</font> between the NotI site and the PstI site
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'''Suffix'''<br>There is <font color='purple'>no T spacer (*)</font> between the Hin coding region and the SpeI site.
 
|}
 
|}
 
<font size='4'>5'</font size><font color='blue'>GAATTC</font color><font color='pink'> GCGGCCGC</font color><font color='gold'> T</font color> <font color='orange'> TCTAGA</font color><font color='red'> -</font color> <u>aatccagctccaccgggccgaggtacgtggcgctgcgttgcgcccctccgtctgttccatatcccgccgcggatgttaggtacggttgggcaaggtacac
 
gagcggctccgccgtatttagcggcactgctagtcgccaggttactagcttcaatccgaccattctcggcgctcgctaggaacttcgacagggactacca
 
gcagtagatggacggacctgtcgtaccggacgttgcgcccgtagggctacggcggccttcgctcttcttagtattaccccttccggtaggtcggagcgca
 
gcgcttgcggtcgttctgcatcgggtcgcgcagccggcggtacggccgctattaccggacgaagagcggctttgcaaaccaccgccccggtcactgcttc
 
cgaactcgctcccgcacgttctaaggcttatggcgttcgctgtccggctagtagcagcgcgaggtcgctttcgccaggagcggcttttactgggtctcgc
 
gacggccgtggacaggatgctcaacgtactatttcttctgtcagtattcacgccgctgctatcagtacggggcgcgggtggccttcctcgactgacccaa
 
cttccgagagttcccgtagccagctgcgagagggaatacgctgaggacgtaatccttcgtcgggtcatcatccaactccggcaactcgtggcggcggcgt
 
tccttaccacgtacgttcctctaccgcgggttgtcagggggccggtgccccggacggtggtatgggtgcggctttgttcgcgagtactcgggcttcaccg
 
ctcgggctagaaggggtagccactacagccgctatatccgcggtcgttggcgtggacaccgcggccactacggccggtgctacgcaggccgcatctccta
 
ggtgtcctgcccacaccagcggtactagcgcatcagctatcaccgaggttcatcgcttcgctcgtcctgacccgccgccggtttcgccagcctgtcacga
 
ggctcttgcccacgcgtatctttaacgtagttgcgtatatcgcgatcgtcgtgcggtatcactgaccgctacgacagccttacctgctatagggcgttct
 
ccgggccgtcatggccgtattggttcggatacggatgtcgtaggtcccactgccacggctcctactgctactcgcgtaacaatctaaagta</u><font color='purple'> -</font color><font color='cyan'> ACTAGT</font color><font color='gold'> A</font color> <font color='magenta'>GCGGCCG</font color><font color='lime green'> CTGCAG</font color>
 
 
<font color='blue'> EcoRI </font color> <font color='pink'> NotI</font color><font color='gold'> Spacer</font color><font color ='orange'> XbaI</font color> <font color='red'> Missing G spacer</font color><u> Coding Region</u> <font color='purple'> Missing T spacer</font color><font color='cyan'> SpeI</font color> <font color='gold'> Spacer</font color> <font color='magenta'> NotI</font color><font color='lime green'> PstI</font color>
 
 
We compared our BioBricks with those from Tom Knight's paper,<u> Idempotent Vector Design for Standard Assembly of Biobricks</u>. As seen below
 
 
[[Image:BioBricks_from_paper.png]]
 
  
 
===Source===
 
===Source===
  
PsB1AT3
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The tetracycline resistance coding region was PCR amplified from <partinfo>pSB1AT3</partinfo>.
  
 
===References===
 
===References===
[https://dspace.mit.edu/handle/1721.1/21168|  Knight, Tom. Idempotent Vector Design for Standard Assembly of Biobricks]
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* [https://dspace.mit.edu/handle/1721.1/21168|  Knight, Tom. Idempotent Vector Design for Standard Assembly of Biobricks]

Revision as of 17:14, 29 October 2006

tetracycline resistance protein TetA(C) (backwards) [cf. BBa_J31007]


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1043
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 897
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 343
    Illegal NgoMIV site found at 503
    Illegal NgoMIV site found at 871
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

BBa J31006 was cloned into vector pSB1A2. The Biobricks on this part are not wild type, but the cut sites are still viable.

Standard BioBrick Cloning Sites (Knight) 5'--GAATTC GCGGCCGC T TCTAGA G ----insert---- T ACTAGT A GCGGCCG CTGCAG--
3'--CTTAAG CGCCGGCG A AGATCT C -------------- A TGATCA T CGCCGGC GACGTC--
BBa_J31001 Cloning Sites 5'--GAATTC GCGGCCGC T TCTAGA * --Tet coding-- * ACTAGT A GCGGCCG CTGCAG--
3'--CTTAAG CGCCGGCG A AGATCT * -------------- * TGATCA T CGCCGGC GACGTC--


Prefix
There is no G spacer (*) between the XbaI and the Hin coding region.
Suffix
There is no T spacer (*) between the Hin coding region and the SpeI site.

Source

The tetracycline resistance coding region was PCR amplified from pSB1AT3.

References