Team:Marburg/Part Collection

P A R T   C O L L E C T I O N

We proudly present the Marburg Collection 2.0, consisting of the Marburg Collection from last year’s iGEM Team and the numerous additions that were made this year.

The Marburg Collection 1.0 is a Golden Gate based toolbox containing 123 parts, including promoters, reporters, fluorescence and epitope tags, oris, resistance cassettes and genome engineering tools. A specialty in this toolbox are the 36 connectors with which multigene constructs can be easily and flexibly cloned. They also allow the inversion of individual transcription units and function as insulators to avoid undesired crosstalk.
The Marburg Collection 2.0 adds placeholders to our arsenal, which allow the large scale assembly of plasmids for screenings and characterizations as well as another addition we called the Green Expansion.

This expansion features the world's first MoClo compatible shuttle vector for cyanobacteria (K3228069). This composite part contains the origins of replication ColE1 (for cloning) and the ori of panS (for maintenance in cyanobacteria) as well as a spectinomycin cassette. It can be used in the Marburg Collection like any other composite part of the type 7+8 (antibiotic cassette + origin of replication).
This expansion also contains all the parts needed for the genomic integration of one or multiple genes in cyanobacteria. It convinces with a striking flexibility and a very intuitive workflow for the de novo assembly of your plasmid of choice. It encompasses five different neutral integration sites to choose from: three conventional sites frequently used in the cyanobacterial community (NSI to NSIII) as well as our own rationally designed artificial neutral integration site options (a.N.S.o. 1 and 2). These sites show no transcriptional activity from neighboring regions and are therefore completely orthogonal. Additionally we offer 4 different antibiotic markers to use (chloramphenicol, gentamycin, spectinomycin and kanamycin). In theory with this setup up to 20 genes could be introduced into a cyanobacterial strain.
Thanks to the flexible design this expansion can also be used for the genomic modification of any chassis after the introduction of new species specific LVL 0 integration sites to our Marburg Collection 2.0. As the workflow to build new homologies is a bit more intricate compared to the one pot one step assembly of our other parts due to the internal BsmbI cutting site, we described the workflow for that in our design section.

The Green Expansion proves a valuable addition to our Marburg Collection 2.0 and to the iGEM Registry of Parts. It services users of our chassis and other cyanobacterial strains with a useful tool for genomic modifications but it also contributes a shell that can be used to modify any other model organism as well.

Parts of the Marburg Collection 2.0

Download all 5'Connectors

BioBrick Name
K3228000 aNSo1 integration up
K3228001 aNSo2 integration up
K3228020 NS1 up
K3228021 NS2 up
K3228022 NS3 up

Download all 3'Homology Connectors

BioBrick Name
K3228002 aNSo1 integration down
K3228003 aNSo2 integration down
K3228023 NS1 down
K3228024 NS2 down
K3228025 NS3 down

Download all Promotors

BioBrick Name
K3228053 T7 consensus Promotor

Download all Terminators

BioBrick Name
K3228004 shortTB0010
K3228005 shortTB0015
K3228006 shortTB1002
K3228007 shortTB1003
K3228008 shortTB1004
K3228009 shortTB1005
K3228010 shortTB1006
K3228011 shortTB1007
K3228012 shortTB1008
K3228013 shortTB1009
K3228014 shortTB1010
K3228015 shortDummy

Download all Antibiotic Resistances

BioBrick Name
K3228016 SpecRes_short
K3228017 CmlRes_short
K3228018 TetRes_short
K3228019 KanRes_short
K3228027 GenRes_short

Download all Shuttle Vectors

BioBrick Name
K3228026 oriT integration
K3228069 panS SpecRes LVL1
K3228089 panS KanRes LVL2

Download all Coding Sequences

BioBrick Name
K3228040 NanoLuc-K1159001
K3228041 NanoLuc (S.e.)
K3228042 TeLuc (S.e.)
K3228043 Antares2 (S.e.)
K3228044 eYFP-E0030
K3228045 sYFP2 (S.e.)
K3228046 pHlourin2 (S.e.)
K3228047 rxYFP (S.e.)
K3228048 sfGFP (S.e.)
K3228049 mTurquoise2 (S.e.)
K3228050 T7-Polymerase (S.e.)
K3228051 Limonene Synthase
K3228052 Farnesen Synthase
K3228100 cpf1

Download all Ribosome-Binding Sites

BioBrick Name
K3228054 riboJ B0034
K3228055 riboJ B0034 noScar
K3228056 SarJ B0034
K3228057 PlmJ B0034

Download all Placeholders

BioBrick Name
K3228060 PRO placeholder.
K3228061 RBS placeholder
K3228062 CDS placeholder
K3228063 TER placeholder

Download all Standard Measurement Vectors

BioBrick Name
K3228073 PRO S.e.
K3228074 RBS S.e.
K3228075 CDS S.e.
K3228090 Ter S.e.

Download the CRISPR-Kit

BioBrick Name
K3228101 crRNA-GFP dropout
K3228103 cpf1+crRNA-GFP dropout