Team:Leiden/Parts

iGEM Leiden | 2019

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

Suckerin Polymer Layer to Achieve Sustainable Health

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Description Demonstrate Model Parts Results

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