iGEM 2013 — Team Tianjin

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# Team:Tianjin/Notebook

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* [Attributions
  &
  Contributions](https://2013.igem.org/Team%3ATianjin/Attributions)

* [June,2013](https://2013.igem.org/Team%3ATianjin/Notebook)
  + | Sun | Mon | Tue | Wed | Thu | Fri | Sat |
    | --- | --- | --- | --- | --- | --- | --- |
    |  |  |  |  |  |  | 1 |
    | 2 | 3 | 4 | 5 | 6 | [7](https://2013.igem.org/Team%3ATianjin/Notebook#anchor-n-01) | 8 |
    | 9 | 10 | 11 | 12 | 13 | 14 | 15 |
    | 16 | 17 | 18 | 19 | [20](https://2013.igem.org/Team%3ATianjin/Notebook#anchor-n-02) | 21 | 22 |
    | 23 | 24 | 25 | [26](https://2013.igem.org/Team%3ATianjin/Notebook#anchor-n-03) | 27 | 28 | 29 |
    | 30 |  |  |  |  |  |  |
* [July,2013](https://2013.igem.org/Team%3ATianjin/Notebook)
  + | Sun | Mon | Tue | Wed | Thu | Fri | Sat |
    | --- | --- | --- | --- | --- | --- | --- |
    |  | 1 | 2 | 3 | 4 | [5](https://2013.igem.org/Team%3ATianjin/Notebook#anchor-n-04) | 6 |
    | 7 | 8 | [9](https://2013.igem.org/Team%3ATianjin/Notebook#anchor-n-05) | 10 | 11 | 12 | 13 |
    | 14 | 15 | 16 | 17 | 18 | 19 | 20 |
    | 21 | [22](https://2013.igem.org/Team%3ATianjin/Notebook#anchor-n-06) | 23 | 24 | 25 | 26 | 27 |
    | 28 | 29 | 30 | 31 |  |  |  |
* [August,2013](https://2013.igem.org/Team%3ATianjin/Notebook)
  + | Sun | Mon | Tue | Wed | Thu | Fri | Sat |
    | --- | --- | --- | --- | --- | --- | --- |
    |  |  |  |  | [1](https://2013.igem.org/Team%3ATianjin/Notebook#anchor-n-07) | 2 | 3 |
    | 4 | 5 | 6 | 7 | 8 | 9 | [10](https://2013.igem.org/Team%3ATianjin/Notebook#anchor-n-08) |
    | 11 | 12 | [13](https://2013.igem.org/Team%3ATianjin/Notebook#anchor-n-09) | 14 | 15 | 16 | 17 |
    | [18](https://2013.igem.org/Team%3ATianjin/Notebook#anchor-n-10) | 19 | 20 | 21 | 22 | [23](https://2013.igem.org/Team%3ATianjin/Notebook#anchor-n-03) | 24 |
    | 25 | 26 | 27 | 28 | 29 | 30 | 31 |
* [Human Practice Notebook](https://2013.igem.org/wiki/index.php?title=Team:Tianjin/Notebook/HP)
* [![](https://static.igem.org/mediawiki/2013/7/75/IGEM-logo-blue.png)](https://2013.igem.org/Main_Page "Main Page")

|  |
| --- |
| Contents   * [1 June 7th, 2013](#June_7th.2C_2013) * [2 June 20th, 2013](#June_20th.2C_2013) * [3 June 26th, 2013](#June_26th.2C_2013) * [4 July 5th, 2013](#July_5th.2C_2013) * [5 July 9th, 2013](#July_9th.2C_2013) * [6 July 22nd, 2013](#July_22nd.2C_2013) * [7 August 1st, 2013](#August_1st.2C_2013) * [8 August 10th, 2013](#August_10th.2C_2013) * [9 August 13th, 2013](#August_13th.2C_2013) * [10 August 18th, 2013](#August_18th.2C_2013) * [11 August 23rd, 2013](#August_23rd.2C_2013) |

# June 7th, 2013

---

Synthesized J23100/J23103-AlkR-tt-PalkM (tt=B1006) directly through pEASY-BLUNT plasmids, respectively.

[![gene](/wiki/images/3/35/Tju-notebook-1.1.jpg)](/File%3ATju-notebook-1.1.jpg "gene")

[![gel 1](/wiki/images/7/7f/Tju-gel-1.1.1.jpg)](/File%3ATju-gel-1.1.1.jpg "gel 1")

**Figure 1.** AlkR-1 ~600bp cut from AlkR-1-pEASY-BLUNT

[![gel 2](/wiki/images/c/c1/Tju-gel-1.1.2.jpg)](/File%3ATju-gel-1.1.2.jpg "gel 2")

**Figure 2.** AlkR-2 ~600bp cut from AlkR-2-pEASY-BLUNT

[![gel 3](/wiki/images/9/9b/Tju-gel-1.1.3.jpg)](/File%3ATju-gel-1.1.3.jpg "gel 3")

**Figure 3.** J23100/J23103-AlkR-tt-RFP ~2000bp cut from J23100/J23103-AlkR-tt-RFP-pEASY-BLUNT

# June 20th, 2013

---

Synthesized J23100/J23103-AlkR-tt-PalkM-RFP/tetA and linked them with pSB1C3, respectively.

[![gene](/wiki/images/b/b3/Tju-notebook-1.2.1.jpg)](/File%3ATju-notebook-1.2.1.jpg "gene")

[![gene](/wiki/images/e/eb/Tju-notebook-1.2.2.jpg)](/File%3ATju-notebook-1.2.2.jpg "gene")

[![gene](/wiki/images/f/f8/Tju-notebook-1.2.3.jpg)](/File%3ATju-notebook-1.2.3.jpg "gene")

[![gene](/wiki/images/8/8c/Tju-notebook-1.2.4.jpg)](/File%3ATju-notebook-1.2.4.jpg "gene")

[![gel 4](/wiki/images/2/2b/Tju-gel-1.2.1.jpg)](/File%3ATju-gel-1.2.1.jpg "gel 4")

**Figure 4.** J23100/J23103-AlkR-tt-PalkM-RFP ~2000bp cut from J23100/J23103-AlkR-tt-PalkM-RFP-pSB1C3

[![gel 5](/wiki/images/6/6d/Tju-gel-1.2.2.jpg)](/File%3ATju-gel-1.2.2.jpg "gel 5")

**Figure 5.** J23100/J23103-AlkR-tt-PalkM-tetA ~2400bp cut from J23100/J23103-AlkR-tt-PalkM-tetA-pSB1C3

# June 26th, 2013

---

Synthesized J23100/J23114-NPDC-AAR with BamHI, SacI and KpnI recognition sites inserted through overlap PCR and linked them with pSB3K3, respectively.

[![gene](/wiki/images/e/eb/Tju-notebook-1.3.jpg)](/File%3ATju-notebook-1.3.jpg "gene")

[![gel 6](/wiki/images/8/8d/Tju-gel-1.3.jpg)](/File%3ATju-gel-1.3.jpg "gel 6")

**Figure 6.** J23100/J23114-NPDC-AAR ~2000bp cut from J23100/J23114-NPDC-AAR-pSB3K3

# July 5th, 2013

---

Constructed J23100/J23114-NPDC-AAR-pSB1K3, respectively.

[![gene](/wiki/images/e/ec/Tju-notebook-1.4.jpg)](/File%3ATju-notebook-1.4.jpg "gene")

[![gel 7](/wiki/images/7/76/Tju-gel-1.4.jpg)](/File%3ATju-gel-1.4.jpg "gel 7")

**Figure7.** J23100/J23114-NPDC-AAR ~2000bp cut from J23100/J23114-NPDC-AAR-pSB1K3

# July 9th, 2013

---

Translated E. coli BL21 with plasmids constructed.

# July 22nd, 2013

---

Acquired YbhGFSR through PCR and linked it with pSB3C5.

[![gene 8](/wiki/images/a/a0/Tju-notebook-1.6.jpg)](/File%3ATju-notebook-1.6.jpg "gene 8")

[![gel 8](/wiki/images/4/48/Tju-gel-1.6.jpg)](/File%3ATju-gel-1.6.jpg "gel 8")

**Figure 8.** YbhGFSR ~5000bp cut from YbhGFSR- pSB3C5

# August 1st, 2013

---

Constructed J23100-RBS（B0030）-YbhGFSR-pSB1C3 （BBa\_K081005）.

[![gene](/wiki/images/e/e6/Tju-notebook-1.7.jpg)](/File%3ATju-notebook-1.7.jpg "gene")

[![gel 9](/wiki/images/7/78/Tju-gel-1.7.jpg)](/File%3ATju-gel-1.7.jpg "gel 9")

**Figure 9.** J23100-RBS（B0030）-YbhGFSR ~5000bp cut from J23100-RBS（B0030）-YbhGFSR-pSB1C3（BBa\_K081005）

# August 10th, 2013

---

Constructed J23100-RBS（B0030）-YbhGFSR-pSB3C5.

[![gene 10](/wiki/images/0/00/Tju-notebook-1.8.jpg)](/File%3ATju-notebook-1.8.jpg "gene 10")

[![gel 10](/wiki/images/d/d1/Tju-gel-1.8.jpg)](/File%3ATju-gel-1.8.jpg "gel 10")

**Figure 10.** J23100-RBS（B0030）-YbhGFSR ~5000bp cut from J23100-RBS（B0030）-YbhGFSR-pSB3C5

# August 13th, 2013

---

Constructed J23100/J23103-AlkR-tt-pSB1C3 and PalkM-RFP/tetA-tt-pSB1C3 (tt=B0015), respectively.

[![gene](/wiki/images/7/71/Tju-notebook-2.1.1.jpg)](/File%3ATju-notebook-2.1.1.jpg "gene")

[![gene](/wiki/images/4/45/Tju-notebook-2.1.2.jpg)](/File%3ATju-notebook-2.1.2.jpg "gene")

[![gene](/wiki/images/b/b9/Tju-notebook-2.1.3.jpg)](/File%3ATju-notebook-2.1.3.jpg "gene")

[![gel 11](/wiki/images/8/8c/Tju-gel-2.1.1.jpg)](/File%3ATju-gel-2.1.1.jpg "gel 11")

**Figure 11.** J23100/J23103-AlkR-tt ~1200bp cut from J23100/J23103-AlkR-tt-pSB1C3

[![gel 12](/wiki/images/4/45/Tju-gel-2.1.2.jpg)](/File%3ATju-gel-2.1.2.jpg "gel 12")

**Figure 12.** PalkM-RFP-tt ~1000bp cut from PalkM-RFP-tt-pSB1C3

[![gel 13](/wiki/images/a/ad/Tju-gel-2.1.3.jpg)](/File%3ATju-gel-2.1.3.jpg "gel 13")

**Figure 13.** PalkM-tetA-tt ~1500bp cut from PalkM-tetA-tt-pSB1C3

# August 18th, 2013

---

Constructed PalkM-RFP/tetA-tt-J23100/J23103-ALKR-tt-pSB1C3, respectively.

[![gene](/wiki/images/3/30/Tju-notebook-2.2.1.jpg)](/File%3ATju-notebook-2.2.1.jpg "gene")

[![gene](/wiki/images/8/82/Tju-notebook-2.2.2.jpg)](/File%3ATju-notebook-2.2.2.jpg "gene")

[![gel 14](/wiki/images/b/b4/Tju-gel-2.2.1.jpg)](/File%3ATju-gel-2.2.1.jpg "gel 14")

**Figure 14.** PalkM-RFP -tt- J23100/J23103-ALKR-tt ~2300bp cut from PalkM-RFP -tt- J23100/J23103-ALKR-tt-pSB1C3

[![gel 15](/wiki/images/1/1e/Tju-gel-2.2.2.jpg)](/File%3ATju-gel-2.2.2.jpg "gel 15")

**Figure 15.** PalkM-tetA -tt- J23100/J23103-ALKR-tt ~2800bp cut from PalkM-tetA-tt- J23100/J23103-ALKR-tt-pSB1C3

# August 23rd, 2013

---

Constructed PalkM-RFP/tetA-tt-J23100/J23103-ALKR-tt-pSB4C5, respectively.

[![gene](/wiki/images/f/fd/Tju-notebook-2.3.1.jpg)](/File%3ATju-notebook-2.3.1.jpg "gene")

[![gene](/wiki/images/3/3c/Tju-notebook-2.3.2.jpg)](/File%3ATju-notebook-2.3.2.jpg "gene")

[![gel 16](/wiki/images/f/fd/Tju-gel-2.3.jpg)](/File%3ATju-gel-2.3.jpg "gel 16")

**Figure 16.** PalkM-RFP -tt- J23100/J23103-ALKR-tt ~2300bp cut from PalkM-RFP -tt- J23100/J23103-ALKR-tt-pSB4C5

E-mail：austinamens@gmail.com | Address:Building No.20, No.92 Weijin Road, Tianjin, China | Zip-code：300072

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