Team:Leiden/Parts

iGEM Leiden | 2019

S.P.L.A.S.H.

Suckerin Polymer Layer to Achieve Sustainable Health

Parts

This year, the iGEM Leiden team decided to make customizable hydrogels from suckerin, a squid-derived protein. Using genetic engineering, suckerin was produced within E. coli and S. cerevisiae. The proteins and constructs for the production and functionality of the hydrogel were assembled in the biobrick standard (RFC25). We have been able to create 7 biobricks of suckerin proteins in several organisms. Furthermore, we have created composite parts of the suckerins under control of the PLac expression cassette. This allowed expression of the suckerin proteins after induction with IPTG. Eventually, all biobricks were used to produce the suckerin proteins. An overview of all submitted biobricks synthesized within this project is given below. Further, this page displays the primers used during this project. Visit our experiments page to see what we did with these parts, and our results page to see what came out of it.

Parts

Name

Type

Description

Designer

Length

Favorite

BBa_K3041017

Composite

PLac-suckerin-12

Jo-Anne Verschoor

2343

BBa_K3041002

Coding

Suckerin-12

Jo-Anne Verschoor

697

BBa_K3041015

Composite

PLac-suckerin-8

Jo-Anne Verschoor

2124

BBa_K3041016

Composite

PLac-suckerin-9

Jo-Anne Verschoor

2214

BBa_K3041000

Coding

Suckerin-8

Jo-Anne Verschoor

480

BBa_K3041001

Coding

Suckerin-9

Melanie Ofman

568

BBa_K3041003

Coding

Suckerin-19

Jo-Anne Verschoor

1200

BBa_K3041004

Coding

Suckerin-8 for yeast

Jo-Anne Verschoor

480

BBa_K3041005

Coding

Suckerin-9 for yeast

Melanie Ofman

567

BBa_K3041006

Coding

Suckerin-12 for yeast

Melanie Ofman

696

BBa_K3041007

Coding

Suckerin M1

Jo-Anne Verschoor

42

BBa_K3041008

Coding

Suckerin M2

Melanie Ofman

72

BBa_K3041009

Composite

M1-M1 module combination

Jo-Anne Verschoor

42

BBa_K3041010

Composite

M1-M2 module combination

Melanie Ofman

122

BBa_K3041011

Composite

M2-M2 module combination

Jo-Anne Verschoor

72

BBa_K3041012

Other

V8 linker

Jo-Anne Verschoor

39

BBa_K3041013

Other

SplB Linker

Jo-Anne Verschoor

30

BBa_K3041014

Plasmid

ipET28

Jo-Anne Verschoor

5220

Primers

Name

Sequence

fwpsing


GAATTCGCGGCCGCTTCTAGAGTCATAAAAAATTTATTTGCTTTGCAAGGGGGG


rvpsing


TACTAGTAGCGGCCGCTGCAGGGATCCGTGATGGTGATGGT


fwSRT19


GAATTCGCGGCCGCTTCTAGAGATTTTGCCAGCGGCAACATC


rvSTR19


TACTAGTAGCGGCCGCTGCAGTTAGTGGAGGAGACCATATC


fwpT7lac


TACTAGTAGCGGCCGCTGCAGCATGGTATATCTCCTTCTTA


rvT7lac


GAATTCGCGGCCGCTTCTAGAGATTTTGCCAGCGGCAACATC


Module 1 [82 bp]

Biobrick_RFC25-Pre-Suf_-A'G'TG_Suckerin iGEM_M1_BamHI_fw


GAATTCGCGGCCGCTTCTAGAGTGGCCGGCCCGGCGGCAACCGCTGT
ATCTCATACCACTCACCATGCACCAGGATCCATGC


Biobrick_RFC25-Pre-Suf_-A'G'TG_Suckerin iGEM_M1_BamHI_rv


GCATGGATCCTGGTGCATGGTGAGTGGTATGAGATACAGCGGTTGC
CGCCGGGCCGGCCACTCTAGAAGCGGCCGCGAATTC


Module 2 [112 bp]

Biobrick_RFC25-Pre-Suf_-A'G'TG_Suckerin iGEM_M2_BamHI_fw


GATCGGATCCGGTTACGGAGGACTTTACGGTGGCTACGGCCTTGGTGGCTTG
TATGGTGGATATGGGCTGGGCTATGGCTATACCGGTTAATACTAGTAGCGGCCGCTGCAG


Biobrick_RFC25-Pre-Suf_-A'G'TG_Suckerin iGEM_M2_BamHI_rv


CTGCAGCGGCCGCTACTAGTATTAACCGGTATAGCCATAGCCCAGCCCATATC
CACCATACAAGCCACCAAGGCCGTAGCCACCGTAAAGTCCTCCGTAACCGGATCCGATC


suck12_NdeI_fw


5'- CAACTGCATATGTACCACGCGAACCACGTC


suck1_NdeI_fw


5'-CAACTGCATATGGCCCTGACGTACGG


suck6_NdeI_fw


5'- CAACTGCATATGTTCCCCGGCTTCATGG


suck8_NdeI_fw


5'- CAACTGCATATGATCTCGCACGGCAGC


suck-strep_rv


5'- CTGCAGCGGCCGCTACTAG


fvaprabiobrick 


5'- GAATTCGCGGCCGCTTCTAGAGCTGACGCCGTTGGATACACC


rvaprabiobrick


5'- TAGTGGATCCCTGCAGCGGCCGCTACTAGTAACTCCACCGCTGATGACATC


Standard Biobrick primers [5’->3’]

Prefix_RFC10-25_Fw 


GAATTCGCGGCCGCTTCTAGA


Suffix_RFC10_Rv


CTGCAGCGGCCGCTACTAGTA


Suck12_Prefix_A'G'TG_FW 


GAATTCGCGGCCGCTTCTAGAGTGGTTCTGCCTG


Suck12_Suffix_RFC10-to-RFC25_AgeI_Rv


CTGCAGCGGCCGCTACTAGTAATTAACCGGTGTGTAAGAAC


Suck1_Prefix-A'G'TG_FW


GAATTCGCGGCCGCTTCTAGAGTGGCATTAACCTATGGCG


Suck1_Suffix_RFC10-RFC25_AgeI_Rv


CTGCAGCGGCCGCTACTAGTAATTAACCGGTATATCCCCCGAAACCAAACCCCC


Suck6_Prefix-A'G'TG_FW


GAATTCGCGGCCGCTTCTAGAGTGTTCCCCGGTTTC


Suck6_Suffix_RFC10-to-RFC25_AgeI_Rv


CTGCAGCGGCCGCTACTAGTAATTAACCGGTGTAATGGCCGAATC


Suck8_Prefix-A'G'TG_FW


GAATTCGCGGCCGCTTCTAGAGTGATTAGTCACGGCAG


Suck8_Suffix_RFC10-to-RFC25_AgeI_Rv


CTGCAGCGGCCGCTACTAGTAATTAACCGGTTTGAAATCCGTAGGGC


Suck9_Prefix-A'G'TG_FW


GAATTCGCGGCCGCTTCTAGAGTGTACCATGCAAAT


Suck9_Suffix_RFC10-to-RFC25_AgeI_Rv


CTGCAGCGGCCGCTACTAGTAATTAACCGGTATTCACCATCTTAATCTC