Davidson Parts

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'''''Hin gene: Part BBa_J31000'''''
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[[Hin gene: Part BBa_J31000]]
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* Used Hin 100-1
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Primers for Hin gene the 72nd base was mutated from a "T" to a "C" in order to remove a naturally occuring SpeI site. The mutatation is in '''bold'''.
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'''Forward primer'''
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5’ gcattctagaatggctactattgggtatattcgggtgtcaacaattgaccaaaatatcgatttacagcgtaatgcgcttac'''C'''agtgcaaattg 3’
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'''Reverse primer'''
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5’ atgcctgcaggcggccgcactagtttaattcattcgtttttttatac  3’
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'''''Hin gene with LVA degredation tag'''''
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Will have the following primers synthesized so that the Hin gene has a LVA degredation tag.
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'''Forward Primer'''
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*Consists of a <font color='green'>BioBrick prefix</font color>, <font color='blue'>four base pairs from plasmid pSB1A2 [http://partsregistry.org/Part:pSB1A2]</font color>, and<font color='orange'> the first three base pairs of the Hin gene</font color>
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<font face="courier new"><font color='blue'>TCTG</font color><font color='green'>GAATTCGCGGCCGCATCTAGAG</font color><font color='orange'>ATG</font color></font face>
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'''''Reverse Primer'''''
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*Consists of the <font color='green'>Bio Brick suffix</font color>,<font color='red'> Stop codon</font color>, <font color='purple'>LVA tag</font color> and the ,<font color='orange'>last 20 base pairs of the Hin gene</font color> (without the stop codon)
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<font face="courier new"><font color='green'>CTGCAGGCGGCCGCTACTAGT</font color><font color='red'>ATT</font color><font color='purple'>AAGCTACTAAAGCGTAGTTTTCGTCGTTTGCAGC</font color><font color='orange'>ATTCATTCGTTTTTTTATAC</font color></font face>
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[[Primers for Hin gene with LVA degredation tag]]
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*Used HinLVA-2
[[Recombinational Enahancer: Part BBa_J3101]]
[[Recombinational Enahancer: Part BBa_J3101]]
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* used RE-11
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'''''Recombinational Enhancer: Part BBa_J3101'''''
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[[HixC]]
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*Used Hix C-2
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These are the oligos that we ordered to make the recombinational enhancer.
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AATTCGCGGCCGCTTCTAGATTCGGGTGTCAACAATTGACCAAAATAT
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CGATTTACAGCGTAATGCGCTTTCTAGTGCAAATTGTGACCGC
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ATTTTACTAGTTGCGGCCGCCTGCA
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GGCGGCCGCAACTAGTAAAATGCGGTCACAATTTGCACTAGAA
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[[Resistance Genes]]
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*Tet Back- used TB1
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AGCGCATTACGCTGTAAATCGATATTTTGGTCAATTGTTGACACCCGAATCTAGAAGCGGCCGCG
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[[Construction Intermediates]]

Latest revision as of 16:11, 31 July 2006

Hin gene: Part BBa_J31000

  • Used Hin 100-1

Primers for Hin gene with LVA degredation tag

  • Used HinLVA-2

Recombinational Enahancer: Part BBa_J3101

  • used RE-11

HixC

  • Used Hix C-2

Resistance Genes

  • Tet Back- used TB1

Construction Intermediates

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