Team:UofUppsala/Parts


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Parts


What would iGEM be without its BioBricks and parts? Here we list the parts we created with some background information.

Basic parts

Amplification

Biobrick number Name SnapGene File
BBa_K3425034 T7 RNA polymerase
BBa_K3425035 T7M5
BBa_K3425036 T7 lys
BBa_K3425037 TEV protease
BBa_K3425038 QB5'
BBa_K3425039 QB3'
BBa_K3425040 QB Replicase
BBa_K3425045 TEV cleavage site
BBa_K3425046 LAA degradation tag
BBa_K3425048 Juniper GFP without ATG

Agglutination

Biobrick number Name SnapGene File
BBa_K3425041 HSP
BBa_K3425042 pelB
BBa_K3425043 B-F10
BBa_K3425044 ehaA

DBDs

Biobrick number Name SnapGene File
BBa_K3425100 DBD-CaFF
BBa_K3425101 pCadBA

CAFF-I

Biobrick number Name SnapGene File
BBa_K3425200 CaFF-I

NarX/NarL

Biobrick number Name SnapGene File
BBa_K3425130 YeaR Regulatory Promoter
BBa_K3425132 GFP - E. coli codon optimized
BBa_K3425138 PydfJ115

Composite parts

Amplification

Biobrick number Name SnapGene File
BBa_K3425051 β-subunit of Qβ replicase with cleavable degradation tag
BBa_K3425052 Qβ5' recognition sequence of Qβ replicase with RBS
BBa_K3425053 Qβ3' recognition sequence of Qβ replicase with terminator
BBa_K3425054 Constitutive expression construct of Juniper GFP
BBa_K3425055 Constitutive expression construct of Juniper GFP
BBa_K3425056 T7 RNA polymerase regulated expression construct of Juniper GFP
BBa_K3425057 IPTG inducible expression construct of Juniper GFP/td>
BBa_K3425059 Juniper GFP template for amplification by Qβ replicase
BBa_K3425060 T7 RNA polymerase regulated Juniper GFP substrate for Qβ replicase/td>
BBa_K3425061 Constitutive expression construct of Qβ replicase
BBa_K3425062 β-subunit of Qβ replicase under control of inducible pCAD promoter
BBa_K3425063 IPTG inducible expression construct of T7 lysozyme
BBa_K3425064 T7 lysozyme under the control of T7 RNA polymerase/td>
BBa_K3425065 T7 lysozyme under the control of T7 RNA polymerase
BBa_K3425069 IPTG inducible expression construct of T7 RNA polymerase

Agglutination

Biobrick number Name SnapGene File
BBa_K3425070 Modular outer membrane display construct for nanobody specific to HSP biomarker
BBa_K3425073 HSP 16.3 with His-tag
BBa_K3425074 Expression construct of HSP 16.3 with His tag

DBDs

Biobrick number Name SnapGene File
BBa_K3425102 DBD-CaFF construct (pSB1C3)
BBa_K3425103 DBD-CaFF construct (pSB3K3)
BBa_K3425104 DBD-CaFF construct (pSB3C11-J23100)
BBa_K3425105 DBD-CaFF construct (pSB3C11-J23108)
BBa_K3425106 pCadBA testing construct (pSB3K3)
BBa_K3425107 pCadBA testing construct (pSB1C3)
BBa_K3425108 pCadBA testing construct (pSB1K01)
BBa_K3425109 DBD-CaFF translational unit (J23100)
BBa_K3425058 Juniper GFP inducible expression construct under pCAD promoter

CAFF-I

Biobrick number Name SnapGene File
BBa_K3425201 pSB1C3 with CaFF-I

NarX/NarL

Biobrick number Name SnapGene File
BBa_K3425131 pyeaR-GFP
BBa_K3425140 PydfJ115_GFP

TypeIIS

Biobrick number Name SnapGene File
BBa_K3425071 Multi-dummy of TU-DY cloned to pSB3C11
BBa_K3425072 Multi-dummy of MTU-DY cloned to pSB4K02

iGEM Type IIS part collection

Until very recently, the main assembly standard supported by the iGEM registry was the BioBrick standard RFC[10]. However, iGEM Type IIS RFC[1000] assembly was recently accepted as a registry standard, and multiple Type IIS backbones and parts were present in the 2019 DNA distribution kit. We strongly believe in the efficiency and power of this cloning method since it can assemble multiple parts in one reaction, and we wanted to contribute to the development of this standard so that it can be used comfortably by the iGEM community.

Our parts collection provides four types of Type IIS parts.

  • New backbones: Improved version of pEven Level 2 backbones, along with new pOdd backbones of medium (10-12) and low (~5) copy number for Level 3 assemblies.
  • Templates: Parts which have no biological function, but are instead easy to use blueprints to design Type IIS-compatible parts.
  • Dummy parts: Placeholders when your planned assembly has less than four parts, the amount needed for iGEM Type IIS reactions.
  • RBS: commonly used RBS from the registry which are tailored specifically for iGEM Type IIS assembly.

Below you can find a list of the parts which belong to this collection, as well as buttons which lead to the Registry page of this iGEM Type IIS Standard Collection and the trace files of the sequencing of all these parts.

Collection page Sequencing trace files

Backbones

Biobrick number Name SnapGene File
BBa_K3425001 pSB3C11
BBa_K3425002 pSB3C12
BBa_K3425003 pSB3C13
BBa_K3425004 pSB3C14
BBa_K3425005 pSB3K01
BBa_K3425006 pSB3K02
BBa_K3425007 pSB3K03
BBa_K3425008 pSB3K04
BBa_K3425009 pSB4K01
BBa_K3425010 pSB4K02
BBa_K3425011 pSB4K03
BBa_K3425012 pSB4K04
BBa_K3425013 pSB4A01
BBa_K3425014 pSB4A02
BBa_K3425015 pSB4A03
BBa_K3425016 pSB4A04

Templates

Biobrick number Name SnapGene File
BBa_K3425017 Type IIS standard promoter template
BBa_K3425018 Type IIS standard RBS template
BBa_K3425019 Type IIS standard CDS template
BBa_K3425020 Type IIS standard terminator template

Dummy parts

Biobrick number Name SnapGene File
BBa_K3425021 TU-DY
BBa_K3425022 MTU-DY
BBa_K3425023 pSB1K01-DY
BBa_K3425024 pSB1K02-DY
BBa_K3425025 pSB1K03-DY
BBa_K3425026 pSB1K04-DY
BBa_K3425027 pSB3C11-DY
BBa_K3425028 pSB3C12-DY
BBa_K3425029 pSB3C13-DY
BBa_K3425030 pSB3C14-DY

RBS

Biobrick number Name SnapGene File
BBa_K3425031 wRBS - B0032
BBa_K3425032 mRBS - B0034
BBa_K3425033 sRBS - B0030

Primers

Name Description Sequence
RNAse_HZ1_Fw Extracting 2 regions of the chromosomal RNAse III gene. Needed for a knockout of the native RNAse III. Also used as a positive control for colony PCR ATTCGTGCAGTTATTACAGC
RNAse_HZ1_Rv ATCTCCTTCACTCATAGCCG
pBS1C_Linear_Fw Linearizing the integration vector pBS1C. Needed for Gibson Assembly of PydfJ115 + GFP construct TTTGACAGCTTATCATCGGC
pBS1C_Linear_Rv GTGTGTTTCCATGTGTCCAG
pBS2E_Linear_Fw Linearizing the integration vector pBS2E. Needed for Gibson Assembly of Rewired NarL + VHHNarX construct GCTATATTTGGAGTTGAGCCTC
pBS2E_Linear_Rv CTTAGCGAAAGATGCAGTCG
AmyE1_Fw Colony PCR (confirm transformation of pBS1C plasmid) GAAGCGTTCACAGTTTCGGG
AmyE1_Rv acatgagaattgacaccttgcc
AmyE2_Fw Colony PCR (confirm transformation of pBS1C plasmid) aacggcctcaacctactac
AmyE2_Rv GCGTAAGTCAGGATATCAACC
LacA1_Fw Colony PCR (confirm transformation of pBS2E plasmid) GCCGTCATCTTTATTATGCTGC
LacA1_Rv ctgttgtttgtcggtgaacg
LacA2_Fw Colony PCR (confirm transformation of pBS2E plasmid) atcagatagatgtcagacgcatgg
LacA2_Rw AGAATCCGCCCATATCGAGC
GAF22_Ext_Fw Isolating VHHNarX coding sequence from the Gibson Assembly Fragment n2. Needed for Gibson Assembly of the insert for the plasmid pBS2E. CAGGTACAGCTCGTGGAATC
GAF22_Ext_Rv TGGGTATCACCCTGGACATC
RewL_VHHX_Seq_1 Sequencing Rewired NarL + VHHNarX construct, present in pBS2E plasmid. gacgttcttgccattgctgc
RewL_VHHX_Seq_2 GAAATCCAATTTGAGCGGATG
RewL_VHHX_Seq_3 Sequencing Rewired NarL + VHHNarX construct, present in pBS2E plasmid. GACACAGCCGTTTACTACTG
RewL_VHHX_Seq_4 CATAGTCAAGCAAGCGAAGTC
PydfJ115_GFP_Seq_1 Sequencing PydfJ115 + GFP construct, present in pBS1C plasmid. AAAGGTCATTGTTGACGCGG
PydfJ115_GFP_Seq_2 tgacaccctcgttaatcg
ydfH_HZ_Fw Extracting 2 regions of the chromossomal ydfHI genes. Needed for a knockout of the native ydfHI two-component system. TATAGCCTCTATCGTGCTGC
ydfH_HZ_Rv ATGACAATCATGATCGGCGC
ydfI_HZ_Fw Extracting 2 regions of the chromossomal ydfHI genes. Needed for a knockout of the native ydfHI two-component system. GAACCGGATATCATTCTGATGG
ydfI_HZ_Rv CTGGACTTTACTGTTCGCTC
RNAse_HZ2_Fw Extracting 2 regions of the chromossomal RNAse III gene. Needed for a knockout of the native RNAse III. ATTTATCGGAGCCTTGTACC
RNAse_HZ2_Rv CGGCATACATACTGAAATCCC
pBS1C_Type2S_Fw Deleting a restriction site in pBS1C plasmid which is incompatible with Type2S assembly. Needed to create a PydfJ115 + lacZ construct. GCTCTTCTGCATTTGACAGCTTATCATCGGC
pBS1C_Type2S_Rv GCTCTTCACATGTGTGTTTCCATGTGTCCAG
FWD 1C3 Lin Linearization of pSB1C3 plasmid tgcagtccggcaaaaaag
RVS 1C3 Lin CGCGAATTCCAGAAATCATC
VF2 Sequencing/amplifying BioBrick parts tgccacctgacgtctaagaa
VR attaccgcctttgagtgagc
Primer_seq_PyeaR_GFP_2x_acVHH_NarX_GB1_GB2_FOR Colony PCR for confirmation of insertion of gibson fragment 1 and 2 CAgcgtgcaattagcagacc
Primer_seq_PyeaR_GFP_2x_acVHH_NarX_GB2_GB1_REV Colony PCR for confirmation of insertion of gibson fragment 2 and 1 GAATCCATTTGCAGATACACGGT
"Primer_seq_PyeaR_GFP_2x_acVHH_NarX_GB2_FOR" Colony PCR for confirmation of insertion of gibson fragment 2 GGTTGGTCGTCAGGCATTACG
"Primer_seq_PyeaR_GFP_2x_acVHH_NarX_GB2_REV" CCTGGTTGTCATTCTGCGCTAC
Primer_seq_PyeaR_GFP_2x_acVHH_NarX_GB2_GB3_FOR Colony PCR for confirmation of insertion of gibson fragment 2 and 3 GTCAAGCAAGCGAAGTCG
Primer_seq_PyeaR_GFP_2x_acVHH_NarX_GB3_GB2_REV Colony PCR for confirmation of insertion of gibson fragment 3 and 2 CATGCAAGACAGAGACTGC
Primer_seq_PyeaR_GFP_2x_acVHH_NarX_GB1_REV Colony PCR for confirmation of insertion of gibson fragment 1 gtaatggttgtccggtaacagg
Primer_seq_PyeaR_GFP_2x_acVHH_NarX_GB3_FOR Colony PCR for confirmation of insertion ofgibson fragment 3 AGCAGCTGATCGTTATGGAAG
Primer_amp_genomic_NarX_DH5alpha_E_coli_FOR Extract NarX sequence from DH5alpha genome ATGCTTAAACGTTGTCTCTC
Primer_amp_genomic_NarX_DH5alpha_E_coli_REV TTACTCATGGGTATCTCCTTG
Primer_amp_PyeaR_GFP_2x_acVHH_NarX_GB1_FOR Amplification primer for Gibson Assembly Fragment 1 tccttagctttcgctaagg
Primer_amp_PyeaR_GFP_2x_acVHH_NarX_GB3_REV Amplification primer for Gibson Assembly Fragment 3 CACCTTGCCCTTTTTTGC
Primer_amp_PyeaR_GFP_2x_acVHH_NarX_GB1_REV Amplification primer for Gibson Assembly Fragment 1 actgtacctaggactgagc
Primer_amp_PyeaR_GFP_2x_acVHH_NarX_GB2_FOR Amplification primer for Gibson Assembly Fragment 2 tctctacttagttgacggc
Primer_amp_PyeaR_GFP_2x_acVHH_NarX_GB2_REV Amplification primer for Gibson Assembly Fragment 2 CACGCCATAGAGTAGATGG
Primer_amp_PyeaR_GFP_2x_acVHH_NarX_GB3_FOR Amplification primer for Gibson Assembly Fragment 3 GTACCATCTACTCTATGGCG
LacZ_T2-FW cloning AATGGTCTCTAATGatgaccatgattacggattcactgg
LacZ_T2-RV cloning aatGGTCTCTAAGCttatttttgacaccagaccaactgg
RBS_B0034_T2 cloning AatggtctcTtactaaagaggagaaaAatgTGagaccAat
RBS_B0034_Rev_comp._T2 cloning atTggtctCAcatTtttctcctctttagtaAgagaccatT
J23100_T2 cloning AATGGTCTCTGGAGttgacggctagctcagtcctaggtacagtgctagcTACTTGAGACCAAT
J23100_Rev_comp_T2 cloning ATTGGTCTCAAGTAgctagcactgtacctaggactgagctagccgtcaaCTCCAGAGACCATT
pBAD24_backbone_amplification-fw amplification aaagcttggctgttttggcgg
pBAD24_backbone_amplification-rv amplification tcatgaattcctcctgctagcc
lacZ_into_pBAD24_gibson-fw: cloning tgggctagcaggaggaattcatgaccatgattacggattcactg
lacZ_pBAD24_gibson-rv cloning gccaaaacagccaagcttttatttttgacaccagaccaactgg
Caff-1_or_pSB1C3_amplification-fw amplification TTTCTGGAATTCGCGGCCGCTTCtagag
Caff-I_or_psB1C3_amplification-rv amplification TACTAGTAGCGGCCGCTGCAGTCCGGC
Caff-I_into_psB1C3-Gibson-fw cloning CGCTAAGGATGATTTCTGGAATTCGCGGCCGCT
Caff-I_into_psB1C3-Gibson-rv cloning CCTTGCCCTTTTTTGCCGGACTGCAGCGGCCGC
pSB1C3_Caff-I_linearization-rv cloning tttctcctctttTTATTACAGAGCAGCAGCATGAGAA
pSB1C3_Caff-I_linearization-fw cloning TACTAGTAGCGGCCGCTGCA
amil-CP_amplifying-fw amplification ATGCTGCTGCTCTGTAATAAaaagaggagaaaatgagtgt
amilCP_amplifying-rv amplification CTTGCCCTTTTTTGCCGGAC
Caff-I-sequencing-2-rv sequencing GTTATTACGCTGAACCAGATCG
AmilCP_FS_Fw cloning TAGCGCTCTTCGTCGAATGatgagtgtgatcgctaaacaaatgac
AmilCP_FS_Rv cloning TTACGCTCTTCTCTCGAAGCttattaggcgaccacaggtttg
Caff-I-sequencing-fw sequencing CGTTGGTGTAAACTATACTCCGATTCC
CaFFI_sequencing-rv sequencing TACCCGTACCATGACGGTAATCG
lacZ_sequencing-fw sequencing ccacggccaccgatattatttgcc
lacZ_sequencing-rv sequencing gatggaccatttcggcacagcc
Caff-I-sequencing-2-rv sequencing GTTATTACGCTGAACCAGATCG
T7pol_G_Fw cgtataatgtgtggaattgtgagcg
T7pol_G_Rv gcattgcatattgatgctctcgg
T7polFS-Fw genomic extraction TAGCGCTCTTCGTCGAATGaacacgattaacatcgctaagaacg
T7polFS-Rv genomic extraction TTACGCTCTTCTCTCGAAGCttacgcgaacgcgaagtccg
T7lysFS-Fw genomic extraction TAGCGCTCTTCGTCGAATGGCTCGTGTACAGTTTAAACAACG
T7lysFS-Rv genomic extraction TTACGCTCTTCTCTCGAAGCTTATCCACGGTCAGAAGTGACCAG
ConstProm-J23100_FS-Fw cloning TAGCGCTCTTCGTCGGGAGttgacggctagctcagtcctaggtacagtgc
ConstProm-J23100_FS-Rv cloning TTACGCTCTTCTCTCGAGTAgctagcactgtacctaggactgagctagcc
LacUV5_operator_FS-Fw cloning TAGCGCTCTTCGTCGGGAGTTTACACTTTATGCTTCCGGCTCGTATAATGtgtggaattg
LacUV5_operator_FS-Rv cloning TTACGCTCTTCTCTCGAGTAttgttatccgctcacaattccacaCATTATACGAGCCGGA
sRBS-B0030_FS-Fw cloning TAGCGCTCTTCGTCGTACTattaaagaggagaaaTAGTAAATGCGAGAGAAGAGCGTAA
sRBS-B0030_FS-Rv cloning TTACGCTCTTCTCTCGCATTTACTAtttctcctctttaatAGTACGACGAAGAGCGCTA
T7M5_promoter_FS-Fw cloning TAGCGCTCTTCGTCGGGAGGTGCACGACTCACTATAGGTACTCGAGAGAAGAGCGTAA
T7M5_promoter_FS-Rv cloning TTACGCTCTTCTCTCGAGTACCTATAGTGAGTCGTGCACCTCCCGACGAAGAGCGCTA
wRBS-B0032_FS-Fw cloning TAGCGCTCTTCGTCGTACTtcacacaggaaagTACTAAATGCGAGAGAAGAGCGTAA
wRBS-B0032_FS-Rv cloning TTACGCTCTTCTCTCGCATTTAGTActttcctgtgtgaAGTACGACGAAGAGCGCTA
Seq_L1_L0_F2 sequencing GTGGCGCTTTCTCATAGCTC
Seq_mRFP1_F1 sequencing AATGCGTCTGAAACTGAAAGACGG
Seq_L0_F3 sequencing GCTCGATATCAAATTACGCCC
Seq_mRFP1_R1 sequencing CGTTCCCATTTGAAACCTTCCG
Seq_L2_L0_F3 sequencing ATATCCCAATGGCATCGTAAAG
Seq_L0_R2 sequencing TTATTGGTGAGAATCCAAGCCTC
Seq_L1_F3 sequencing AAACTCACCGAGGCAGTTCC
Seq_L1_R2 sequencing TTGCTCCAGGCCGCGATTAAATTC
Seq_L1_R3 sequencing GAGCATTACGCTGACTTGACG
Seq_L2_F1 sequencing TTATTGCTGATAAATCTGGAGCCG
Seq_L2_F2 sequencing ATCTTTATAGTCCTGTCGGGTTTC
Seq_L2_R2 sequencing ACAAACGGCATGATGAACCTGG
Seq_L2_R3 sequencing CGTGGCTTTGTTGAATAAATCG
Seq_L2_R4 cloning ATCCGGTAACTATCGTCTTGAGTC
TU_dummy_Fw cloning TAGCGGTCTCGGGAGAACACGCTCGAGACCGTAA
TU_dummy_Rev cloning TTACGGTCTCGAGCGTGTTCTCCCGAGACCGCTA
MTU_dummy_Fw cloning TAGCGCTCTTCAATGAACAGGTCGAAGAGCGTAA
MTU_dummy_Rev mutagenesis TTACGCTCTTCGACCTGTTCATTGAAGAGCGCTA
Mut_L2_Rev mutagenesis TCACCGGCTCCAGATTTATC
Mut_L2_Fw fusion PCR GCGTGGGTCTGGCGGTATCAT
Qbeta3'_w_SapI_Fw fusion PCR TAGCgctcttcttcgGCTTggcctgctagacgggc
Qbeta3'_w_SapI_rv fusion PCR TTACGCTCTTCTgcctcaAAGCccccgggtg
Ter_w_SapI_Fw fusion PCR TAGCGCTCTTCgggcatcaaataaaacgaaaggctc
Ter_w_SapI_Rv to get pCAD into T IIS Lv 0 TTACGCTCTTCTCTCaAGCGtataaacgcagaaag
pCAD_T2S_FSL0_Fw to get pCAD into T IIS Lv 0 TAGCGCTCTTCGTCGGGAGTTGTAAACATTAAATGTTTATCTTTTCATGATATCAAC
pCAD_T2S_FSL0_Rev T IIS Bacillus RBS TTACGCTCTTCTCTCGAGTAAGTAGGCGATAAGATTACTCACG
BS_mRBS_R1-8_Fw T IIS Bacillus RBS TAGCGCTCTTCGTCGTACTGCTCTTAAAGGAGATTTTAGAATGCGAGAGAAGAGCGTAA
BS_mRBS_R1-8_Rv TTACGCTCTTCTCTCGCATTCTAAAATCTCCTTTAAGAGCAGTACGACGAAGAGCGCTA
BS_pPS11_T2S_L0_Fw T IIS Bacillus promoter TAGCGCTCTTCGTCGGGAGactgcgtcaatacacgttgacactcttttgagaatatgtg
BS_pPS11_T2S_L0_Rv T IIS Bacillus promoter TTACGCTCTTCTCTCGAGTActgataatatcacatattctcaaaagagtgtcaacgtg
AmilCP_FS_Fw cloning (agglutination) TAGCGCTCTTCGTCGAATGatgagtgtgatcgctaaacaaatgac
AmilCP_FS_Rv cloning (agglutination) TTACGCTCTTCTCTCGAAGCttattaggcgaccacaggtttg
J23108_Fw cloning TAGCGCTCTTCGTCGGGAGctgacagctagctcagtcctaggtataatg
J23108_Rv cloning TTACGCTCTTCTCTCGAGTAgctagcattatacctaggactgagctagct
pelB_SP_FS_Fw cloning (agglutination) TAGCGCTCTTCGTCGAATGAAACGTTTCGCACTTAGCTTATTGGCAGGGCTGGTAG
pelB_SP_FS_Rv cloning (agglutination) TTACGCTCTTCTCTCGCGCTGACGCTTGCAGGGCTACCAGCCCTGCCAATAAGC
Seq_QBeta_fw2 sequencing gcggtgccacaacaacgaatag
Seq_QBeta_rv2 sequencing gtggcgtatgtagaaaggtgtg
Seq_QBeta_fw1 sequencing tgcagctcgttgattgaagtt
Seq_QBeta_rv1 sequencing cgcgaattacgccaccaatg
Seq_JGFP_fw sequencing ccacactgcctgtacatctg
Seq_JGFP_rv sequencing gcgctcttgcacataaccttc
J23104_F cloning TAGCGCTCTTCGTCGGGAGttgacagctagctcagtcctaggtattgtg
J23104_R cloning TTACGCTCTTCTCTCGAGTAgctagcacaatacctaggactgagctagct
J23105_F cloning TAGCGCTCTTCGTCGGGAGtttacggctagctcagtcctaggtactatg
J23105_R cloning TTACGCTCTTCTCTCGAGTAgctagcatagtacctaggactgagctagcc
J23113_F cloning TAGCGCTCTTCGTCGGGAGctgatggctagctcagtcctagggattatgc
J23113_R cloning TAGCGCTCTTCGTCGGGAGctgatggctagctcagtcctagggattatgc
J23114_F cloning TAGCGCTCTTCGTCGGGAGtttatggctagctcagtcctaggtacaatg
J23114_R cloning TTACGCTCTTCTCTCGAGTAgctagcattgtacctaggactgagctagcc
Seq_pSB4A0# sequencing (new plasmid) acattgtcgatctgttcatggtg
Seq_Rep101 sequencing (new plasmid) caccataagcattttccctactgatg
GIBSON_3K#2_Fw_4A#2_Rv_4K#1_Fw gibson (new plasmid) ggctgcggcgagcggtatc
GIBSON_3K#2_Rv gibson (new plasmid) cctcatcagtgccaacatagtaagccagtatac
GIBSON_3K#1_Rv_4A#1_Fw gibson (new plasmid) aatgtcatgataataatggtttcttagacgtcag
GIBSON_3K#1_Fw gibson (new plasmid) gcggagtgtatactggcttactatg
GIBSON_4A#1_Rv_4K#1_Rv gibson (new plasmid) caaatggatctcgtaaccgaacttg
GIBSON_4A#2_Fw_4K#2_Fw gibson (new plasmid) ttatctggttgttctcaagttcggttacgagatcc
GIBSON_4K#2_Rv gibson (new plasmid) aatgtcatgataataatggtttcttagacgtcaggtggc
FWD CR Adding Gibson-related overhangs to C-Read construct (not ordered) AAGGATGATTTCTGGAATTCGCG
REV CR AGGACTGAGCTAGCCGTCAATTATTATTTGTACAGTTCGTCCATACCG
FWD LIN 1C3-DBD Linearization of pSB1C3 + DBD-CaFF with GFP overhang ACGAACTGTACAAATAATAATTGACGGCTAGCTCAGTCCTAG/td>
REV LIN 1C3 CGCGAATTCCAGAAATCATC
FWD seqDBD-CaFF pSB1C3 + DBD-CaFF sequencing CACTCTCTGAACATCCAGAACAC
REV seqDBD-CaFF CGGACCACCACGTTTAGATT
FWD seqCRead pSB1C3 + C-Read sequencing CTGTCTAAAGACCCGAACGA
REV seqCRead CCGCACGGGTTTTGTAGTTA
FWD LIN 3K3-mRFP1 Linearizing primers of pSB3K3 ATGGCTTCCTCCGAAGACGTT
REV LIN 3K3-mRFP1 GGCCGCGAATTCCAGAAAT
REV seqpCadBA Sequencing primers of plasmid pSB3K3 with pCadBA GATAACGTCTTCGGAGGAAGC
A lin DBD PSB1C3 FWD Linearizing primers for pSB1C3-DBD-Caff tggaaacgtatcgctaataaATGCAGCAGCCGGTT
B lin DBD PSB1C3 FWD Linearizing primers for pSB1C3-DBD-Caff tggaaacgtatcgctaataaATGCAACAGCCGGTTGTTC
lin DBD PSB1C3 REV Linearizing primers for pSB1C3-DBD-Caff aacagtgcaccagacgacatCTAGTATTTCTCCTCTTTCTCTAGTAGC
JGFP FWD OH Adding JGFP overhang primers (to DBD-Caff) GAGAAAGAGGAGAAATACTAGatgtcgtctggtgcac
JGFP REV OH ACGAACAACCGGCTGCTGCATttattagcgatacgtttccag
JGFP Isol FWD Isolating (and linerazing) primers for JGFP in plasmid pSB1C00+BBa_J97005 atgtcgtctggtgcactg
JGFP Isol REV ttattagcgatacgtttccagg
DBD-Caff Ampli FW Amplification primers for DBD-Caff TCCTTAGCTTTCGCTAAGG
DBD-Caff Ampli REV CACCTTGCCCTTTTTTGC
FWD AMPL PCAD Amplification primers for pCadBA TCCTTAGCTTTCGCTAAGG
REV AMPL PCAD GGACTCTTTGATAACGTCTTCG
F-DBDTypeII Mutagenic primers for adapting DBD-CaFF to the TypeIIS assembly system TAGCGCTCTTCGTCGA/ATGCAGCAGCCGGTTG
R-DBDTypeII TTACGCTCTTCTCTCGAAGC/CAGCGGCCGCTACTAGTA
REV seq_pCad2 Sequencing primer of pCadBA 2 GTAACGGTAACAACACCAC
F-mutProm Mutagenic primers for promoter J23100 (target J23117) GNANAGTGCTAGCTACTAGAGAA
R-mutProm CTAGGACTGAGCTAGCNGTCAA
F-mutProm2 Mutagenic primers for promoter J23100 (target J23117) GGNattatgctagcTACTAGAGAAAG
R-mutProm2 TAGGACTGAGCTAGCCNTCAN

Disclaimer

Please note that 3 of our basic parts (DBD-Caff, VHH-NarX, and Rewired NarL) and all Qβ parts are covered by patents mentioned in the following papers:

Name Reference
DBD-Caff Chang, H.-J., Mayonove, P., Zavala, A., De Visch, A., Minard, P., Cohen-Gonsaud, M., and Bonnet, J. (2018) A Modular Receptor Platform To Expand the Sensing Repertoire of Bacteria. ACS Synth. Biol. 7, 166–175
VHH-NarX Mazé, A., and Benenson, Y. (2020) Artificial signaling in mammalian cells enabled by prokaryotic two-component system. Nat Chem Biol. 16, 179–187
Rewired NarL Schmidl, S. R., Ekness, F., Sofjan, K., Daeffler, K. N.-M., Brink, K. R., Landry, B. P., Gerhardt, K. P., Dyulgyarov, N., Sheth, R. U., and Tabor, J. J. (2019) Rewiring bacterial two-component systems by modular DNA-binding domain swapping. Nat Chem Biol. 15, 690–698
Qβ Yao, Y., Zhang, W., Zhang, M., Jin, S., Guo, Y., Zu, Y., Ren, K., Wang, K., Chen, G., Lou, C., and Wu, Q. (2019) A Direct RNA-to-RNA Replication System for Enhanced Gene Expression in Bacteria. ACS Synth. Biol. 8, 1067–1078

Out of these, iGEM Uppsala 2020 uploaded only the DBD-Caff and the Qβ related parts into the Registry, as their sequences are the only ones provided in the supplementary information of their respective papers