Difference between revisions of "Part:BBa K1926003"
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<partinfo>BBa_K1926003 short</partinfo>
 
<partinfo>BBa_K1926003 short</partinfo>
  
 
This part is a cyclic promoter named CDK4 from human genome. It can lead to transcription of the downstream DNA sequence in every G1 phase[1].
 
This part is a cyclic promoter named CDK4 from human genome. It can lead to transcription of the downstream DNA sequence in every G1 phase[1].
  
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K1926003 SequenceAndFeatures</partinfo>
  
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1. Guo, Z.Y., et al., The elements of human cyclin D1 promoter and regulation involved. Clin Epigenetics, 2011. 2(2): p. 63-76.
 
  
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===Usage===
  
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===You may use this part to:===
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You may use this part to:
  
 
1) Express something in mammal cell lines particularly in G1 phases or once in every cell cycle by stable transfecting it into cell line;
 
1) Express something in mammal cell lines particularly in G1 phases or once in every cell cycle by stable transfecting it into cell line;
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===Source:===
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===Source===
  
 
The sequence was retrieved from Addgene. We got it from human genome through PCR using the following primers:
 
The sequence was retrieved from Addgene. We got it from human genome through PCR using the following primers:
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===Reference===
  
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1. Guo, Z.Y., et al., The elements of human cyclin D1 promoter and regulation involved. Clin Epigenetics, 2011. 2(2): p. 63-76.
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K1926003 SequenceAndFeatures</partinfo>
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Revision as of 08:55, 11 October 2016

A cyclic promoter of CDK4 from human genome

This part is a cyclic promoter named CDK4 from human genome. It can lead to transcription of the downstream DNA sequence in every G1 phase[1].

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 522
    Illegal NheI site found at 526
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 531
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 142
    Illegal AgeI site found at 173
  • 1000
    COMPATIBLE WITH RFC[1000]


Usage

You may use this part to:

1) Express something in mammal cell lines particularly in G1 phases or once in every cell cycle by stable transfecting it into cell line;

2) Use it as a human promoter by transient transfecting it into cells.


Source

The sequence was retrieved from Addgene. We got it from human genome through PCR using the following primers:

pCDK4-F: TGGTGGAGCGAAAAGGTGACAGCATC

pCDK4-R: GCGGAAACTGGGAGGGCGGGGCGAA

(Product: 457)


Promoter function confirmation

G1 promoter function confirmation by transient transfection using 293T cells. Photos taken 48 hours after transient transfection, 10x. pCDK4, pKi67 and pCCNE are our G1 promoters. pmPGK is the constitutive promoter of mouse PGK, it is a medium promoter, here used as a control.

Figure 1: in vivo testing pG1 promoter function in human 293 cell line.



Reference

1. Guo, Z.Y., et al., The elements of human cyclin D1 promoter and regulation involved. Clin Epigenetics, 2011. 2(2): p. 63-76.