Team:Tokyo Tech/Projects/PHAs/detail/index.htm

From 2012.igem.org

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PHA production detail</div>
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__NOTOC__
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<div class="whitebox">
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<div id="tokyotech" style=" font:bold ;left ; font-size: 15px; color: #000000; padding: 10px;">
<div id="tokyotech" style=" font:bold ;left ; font-size: 15px; color: #000000; padding: 10px;">
=Production trial of PHAs by past teams=
=Production trial of PHAs by past teams=
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[[https://2012.igem.org/Team:Tokyo_Tech/Projects/PHAs/index.htm#Construction_of_phaC1-A-B1_in_Biobrick_format Back to "Construction of phaC1-A-B1 in Biobrick format"]]
We introduce some attempts in the past iGEM to show how great our work is in iGEM.  
We introduce some attempts in the past iGEM to show how great our work is in iGEM.  
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| align="center" style="background:#f0f0f0;"|'''experience'''
| align="center" style="background:#f0f0f0;"|'''experience'''
|-
|-
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K338003 BBa_K338003]||PHA Synthase Composite, Part 1/2||planning||- |
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K338003 BBa_K338003]||PHA Synthase Composite, Part 1/2||Planning||
|-
|-
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K338004 BBa_K338004]||PHA Synthase Composite, Part 1/2||Available||None|
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K338004 BBa_K338004]||PHA Synthase Composite, Part 1/2||Available||None
|}
|}
They couldn’t prove that their engineered bacteria produced PHB according to the team wiki.  
They couldn’t prove that their engineered bacteria produced PHB according to the team wiki.  
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| align="center" style="background:#f0f0f0;"|'''experience'''
| align="center" style="background:#f0f0f0;"|'''experience'''
|-
|-
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K342001 BBa_K342001]||Pha C (poly-β-hydroxybutyrate polymerase)||available||None|
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K342001 BBa_K342001]||Pha C (poly-β-hydroxybutyrate polymerase)||Available||None
|}
|}
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They registered and submitted a part, which contains only phaC sequence, as worked. But the team wiki shows that the part doesn’t meet the standard which is required in iGEM because they couldn’t cut their part with Xba1.
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They registered and submitted a part, which contains only phaC sequence, as worked. But the team wiki shows that the part doesn’t meet the standard which is required in iGEM because they couldn’t cut their part with XbaI.
[https://2009.igem.org/Team:Duke iGEM09_Duke]
[https://2009.igem.org/Team:Duke iGEM09_Duke]
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| align="center" style="background:#f0f0f0;"|'''experience'''
| align="center" style="background:#f0f0f0;"|'''experience'''
|-
|-
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K282000 BBa_K282000]||phaAB||available|| |
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K282000 BBa_K282000]||phaAB||Available||None
|}
|}
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[https://2008.igem.org/Team:Utah State iGEM08_Utah State]
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[https://2008.igem.org/Team:Utah_State iGEM08_Utah State]
{| class="wikitable" cellpadding="4"
{| class="wikitable" cellpadding="4"
| align="center" style="background:#f0f0f0;"|'''Part number'''
| align="center" style="background:#f0f0f0;"|'''Part number'''
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| align="center" style="background:#f0f0f0;"|'''experience'''
| align="center" style="background:#f0f0f0;"|'''experience'''
|-
|-
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K089001 BBa_K089001]||phaA gene||planning|| |
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K089001 BBa_K089001]||phaA gene||Planning||
|-
|-
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K089002 BBa_K089002]||phaB gene||planning|| |
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K089002 BBa_K089002]||phaB gene||Planning||
|-
|-
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K089003 BBa_K089003]||phaC gene||planning|| |
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K089003 BBa_K089003]||phaC gene||Planning||
|}
|}
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[https://2008.igem.org/Team:Hawaii iGEM08_Hawaii]
[https://2008.igem.org/Team:Hawaii iGEM08_Hawaii]
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| align="center" style="background:#f0f0f0;"|'''experience'''
| align="center" style="background:#f0f0f0;"|'''experience'''
|-
|-
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K125801 BBa_K125801]||RBS-phaA||available|| |
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K125801 BBa_K125801]||RBS-phaA||Available||None
|-
|-
| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K125802 BBa_K125802]
| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K125802 BBa_K125802]
||RBS-phaB||Planning|||
||RBS-phaB||Planning|||
|-
|-
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K125803 BBa_K125803]||RBS-phaC||Available|| |
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K125803 BBa_K125803]||RBS-phaC||Available||None
|-
|-
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K125804 BBa_K125804]||RBS-phaE||Available|| |
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K125804 BBa_K125804]||RBS-phaE||Available||None
|-
|-
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K125501 BBa_K125501]||phaA BioPlastic polyhydroxybutyrate synthesis pathway (origin PCC6803 slr1994)||Available|| |
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K125501 BBa_K125501]||phaA BioPlastic polyhydroxybutyrate synthesis pathway (origin PCC6803 slr1994)||Available||None
|-
|-
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K125502 BBa_K125502]||phaB BioPlastic polyhydroxybutyrate synthesis pathway (origin PCC6803 slr1994)||Available|| |
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K125502 BBa_K125502]||phaB BioPlastic polyhydroxybutyrate synthesis pathway (origin PCC6803 slr1994)||Available||None
|-
|-
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K125503 BBa_K125503]||phaC BioPlastic polyhydroxybutyrate synthesis pathway (origin PCC6803 slr1994)||Available|| |
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K125503 BBa_K125503]||phaC BioPlastic polyhydroxybutyrate synthesis pathway (origin PCC6803 slr1994)||Available||None
|-
|-
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K125504 BBa_K125504]||phaE BioPlastic polyhydroxybutyrate synthesis pathway (origin PCC6803 slr1994)||Available|| |
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K125504 BBa_K125504]||phaE BioPlastic polyhydroxybutyrate synthesis pathway (origin PCC6803 slr1994)||Available||None
|}
|}
[https://2008.igem.org/Team:Virginia iGEM08_Virginia]
[https://2008.igem.org/Team:Virginia iGEM08_Virginia]
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| align="center" style="background:#f0f0f0;"|'''experience'''
| align="center" style="background:#f0f0f0;"|'''experience'''
|-
|-
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K156031 BBa_K156031]||RBS + phaA + double terminator||available|| |
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K156031 BBa_K156031]||RBS + phaA + double terminator||Available||None
|-
|-
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K156033 BBa_K156033]||RBS + phaB1 + double terminator||available|| |
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K156033 BBa_K156033]||RBS + phaB1 + double terminator||Available||None
|-
|-
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K156034 BBa_K156034]||RBS + phaC1 + double terminator||available|| |
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K156034 BBa_K156034]||RBS + phaC1 + double terminator||Available||None
|-
|-
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|[http://partsregistry.org/wiki/index.php?title=Part:BBa_K156012 BBa_K156012] ||phaA (acetyl-CoA acetyltransferase)||available|||
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|[http://partsregistry.org/wiki/index.php?title=Part:BBa_K156012 BBa_K156012] ||phaA (acetyl-CoA acetyltransferase)||Available||None
|-
|-
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K156013 BBa_K156013]||phaB1 (acetyacetyl-CoA reductase)||available|| |
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K156013 BBa_K156013]||phaB1 (acetyacetyl-CoA reductase)||Available||None
|-
|-
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K156014 BBa_K156014]||phaC1 (Poly(3-hydroxybutyrate) polymerase)||available|| |
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K156014 BBa_K156014]||phaC1 (Poly(3-hydroxybutyrate) polymerase)||Available||None
|-
|-
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K156021 BBa_K156021]||Promoter + RBS + phaA + double terminator||available|| |
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K156021 BBa_K156021]||Promoter + RBS + phaA + double terminator||Available||None
|-
|-
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K156018 BBa_K156018]||Promoter + RBS + phaA||planning|| |
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K156018 BBa_K156018]||Promoter + RBS + phaA||Planning|| |
|-
|-
-
| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K156019 BBa_K156019]||Promoter + RBS + phaB1||planning|| |
+
| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K156019 BBa_K156019]||Promoter + RBS + phaB1||Planning|| |
|-
|-
-
| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K156020 BBa_K156020]||Promoter + RBS + phaC1||planning|| |
+
| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K156020 BBa_K156020]||Promoter + RBS + phaC1||Planning|| |
|-
|-
| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K156022 BBa_K156022]
| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K156022 BBa_K156022]
-
||Promoter + RBS + phaB1 + double terminator||planning|| |
+
||Promoter + RBS + phaB1 + double terminator||Planning|| |
|-
|-
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K156023 BBa_K156023]||Promoter + RBS + phaC1 + double terminator||planning|| |
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| [http://partsregistry.org/wiki/index.php?title=Part:BBa_K156023 BBa_K156023]||Promoter + RBS + phaC1 + double terminator||Planning|| |
|}
|}
-
=Materials & Methods=
 
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==Construction of pha-C1-A-B1 in Biobrick format==
+
[[https://2012.igem.org/Team:Tokyo_Tech/Projects/PHAs/index.htm#Construction_of_phaC1-A-B1_in_Biobrick_format Back to "Construction of phaC1-A-B1 in Biobrick format"]]
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[[File:tokyotech PHA biobrick.png|250px|thumb|right|fig3,construction of phaC1-A-B1]]
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To construct a part that meets Biobrick format, we have modified the phaC1-A-B1 operon not to contain forbidden restriction enzyme sites. First, we cloned the wild type gene phaC1-A-B1 from R.eutropha H16 by using PCR and inserted the gene into pSB1C3. However, wild type phaC1-A-B1 gene sequence contains one NotI and three PstI recognition sites that are not allowed in Biobrick format. To get phaC1-A-B1 sequence without these recognition sites, we ordered the chemically synthesized DNA from IDT/MBL. In this chemically synthesized DNA, coding is optimized for E.coli. We used restriction enzyme XbaI (on pSB1C3) and BsrGI (on phaC1-A-B1) to insert sequence. That is to say, we got Poly[(R)-3-hydroxybutyrate] synthesizing gene in Biobrick format ([http://partsregistry.org/wiki/index.php?title=Part:BBa_K934001 BBa_K934001]).
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Fig3:construction of phaC1-A-B1
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Latest revision as of 09:23, 22 October 2012

bar




Production trial of PHAs by past teams

[Back to "Construction of phaC1-A-B1 in Biobrick format"]

We introduce some attempts in the past iGEM to show how great our work is in iGEM.

iGEM10_Caltech

Part number Description States experience
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K338003 BBa_K338003]PHA Synthase Composite, Part 1/2Planning
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K338004 BBa_K338004]PHA Synthase Composite, Part 1/2AvailableNone

They couldn’t prove that their engineered bacteria produced PHB according to the team wiki.


iGEM10_INSA-Lyon

Part number Description States experience
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K342001 BBa_K342001]Pha C (poly-β-hydroxybutyrate polymerase)AvailableNone

They registered and submitted a part, which contains only phaC sequence, as worked. But the team wiki shows that the part doesn’t meet the standard which is required in iGEM because they couldn’t cut their part with XbaI.

iGEM09_Duke

Part number Description States experience
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K282000 BBa_K282000]phaABAvailableNone

iGEM08_Utah State

Part number Description States experience
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K089001 BBa_K089001]phaA genePlanning
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K089002 BBa_K089002]phaB genePlanning
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K089003 BBa_K089003]phaC genePlanning

iGEM08_Hawaii

Part number Description States experience
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K125801 BBa_K125801]RBS-phaAAvailableNone
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K125802 BBa_K125802] RBS-phaBPlanning
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K125803 BBa_K125803]RBS-phaCAvailableNone
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K125804 BBa_K125804]RBS-phaEAvailableNone
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K125501 BBa_K125501]phaA BioPlastic polyhydroxybutyrate synthesis pathway (origin PCC6803 slr1994)AvailableNone
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K125502 BBa_K125502]phaB BioPlastic polyhydroxybutyrate synthesis pathway (origin PCC6803 slr1994)AvailableNone
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K125503 BBa_K125503]phaC BioPlastic polyhydroxybutyrate synthesis pathway (origin PCC6803 slr1994)AvailableNone
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K125504 BBa_K125504]phaE BioPlastic polyhydroxybutyrate synthesis pathway (origin PCC6803 slr1994)AvailableNone

iGEM08_Virginia

Part number Description States experience
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K156031 BBa_K156031]RBS + phaA + double terminatorAvailableNone
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K156033 BBa_K156033]RBS + phaB1 + double terminatorAvailableNone
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K156034 BBa_K156034]RBS + phaC1 + double terminatorAvailableNone
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K156012 BBa_K156012] phaA (acetyl-CoA acetyltransferase)AvailableNone
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K156013 BBa_K156013]phaB1 (acetyacetyl-CoA reductase)AvailableNone
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K156014 BBa_K156014]phaC1 (Poly(3-hydroxybutyrate) polymerase)AvailableNone
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K156021 BBa_K156021]Promoter + RBS + phaA + double terminatorAvailableNone
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K156018 BBa_K156018]Promoter + RBS + phaAPlanning
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K156019 BBa_K156019]Promoter + RBS + phaB1Planning
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K156020 BBa_K156020]Promoter + RBS + phaC1Planning
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K156022 BBa_K156022] Promoter + RBS + phaB1 + double terminatorPlanning
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K156023 BBa_K156023]Promoter + RBS + phaC1 + double terminatorPlanning


[Back to "Construction of phaC1-A-B1 in Biobrick format"]
Retrieved from "http://2012.igem.org/Team:Tokyo_Tech/Projects/PHAs/detail/index.htm"