Team:Pittsburgh/Parts
Parts
Throughout our research process, we designed and synthesized many recombinant protein constructs. These constructs generally consist of several different domains such as split-intein halves and model exteins as well as standard protein purification and solubility tags.
Our parts are organized into basic and composite parts following iGEM protocols. However, in practice we synthesized full length gene fragments and cloned them into our expression vectors.
Additionally, all of the protein-coding parts are codon-optimized for expression in E. coli.
All parts
Here we have listed parts that were constructed to test our systems. We have entered all the constructs that we have designed for our Nested Intein and Split-Linker system as composite parts consisting fo a series of Basic Parts. The sequences of each intein terminus and extein used was uploaded to the iGEM Part Registry as Basic Parts. All of our construct are meant to be cloned in expression vectors; however we were limited to certain expression vectors based on what vectors DNA synthesis companies have available. We showed preference to expressing our proteins in pTEV5 plasmid backbone.
| Construct | Classification | Part Number | Type |
| pVS00028 | N1 | BBa_K3308007 | composite |
| pVS00029 | N2 | BBa_K3308008 | composite |
| pVS00030 | C1 | BBa_K3308009 | composite |
| pVS00031 | C2 | BBa_K3308010 | composite |
| pVS00043 | NSP | BBa_K3308011 | composite |
| pVS00033 | CSP | BBa_K3308012 | composite |
| pVS00034 | MSP | BBa_K3308013 | composite |
| pVS00020 | N1 | BBa_K3308000 | composite |
| pVS00021 | N2 | BBa_K3308001 | composite |
| pVS00022 | C1 | BBa_K3308002 | composite |
| pVS00023 | C2 | BBa_K3308003 | composite |
| pVS00042 | NSP | BBa_K3308004 | composite |
| pVS00025 | CSP | BBa_K3308005 | composite |
| pVS00026 | MSP | BBa_K3308006 | composite |
| pVS000036 | N1 | BBa_K3308115 | composite |
| pVS000037 | N2 | BBa_K3308116 | composite |
| pVS000038 | C1 | BBa_K3308117 | composite |
| pVS000039 | C2 | BBa_K3308118 | composite |
| pVS000040 | NSP | BBa_K3308119 | composite |
| pVS000041 | CSP | BBa_K3308120 | composite |
| pHG000044 | linker-AB | BBa_K3308027 | composite |
| pHG000045 | linker-AC | BBa_K3308028 | composite |
| pHG000047 | linker-BA | BBa_K3308029 | composite |
| pHG000052 | linker-CB | BBa_K3308030 | composite |
| pHG000059 | N-VMA-A | BBa_K3308031 | composite |
| pHG000060 | N-VMA-B | BBa_K3308032 | composite |
| pHG000063 | C-VMA-A | BBa_K3308033 | composite |
| pHG000064 | C-VMA-B | BBa_K3308034 | composite |
| pHG000067 | cis-A | BBa_K3308035 | composite |
| pHG000068 | cis-AB | BBa_K3308036 | composite |
| All pET-28 composite parts | RBS_pET-28 | BBa_K3308042 | basic |
| All pET-28 composite parts | pET-28 6x His Tag | BBa_K3308046 | basic |
| All pET-28 composite parts | pET-28 Thrombin Site | BBa_K3308080 | basic |
| All pTEV5 composite parts | RBS_ pTEV5 | BBa_K3308062 | basic |
| All pTEV5 composite parts | pTEV5 6x His Tag | BBa_K3308091 | basic |
| All pTEV5 composite parts | pTEV5 Tev Protease Recognition Site | BBa_K3308092 | basic |
| All pKLD66 composite parts | RBS_pKLD66 | BBa_K3308094 | basic |
| All pKLD66 composite parts | pKLD66 Tev Protease Recognition Site | BBa_K3308089 | basic |
| All pKLD66 composite parts | Maltose binding protein (malE) | BBa_K3308090 | basic |
| BBa_K3308000 | gpd-GGGIG | BBa_K3308043 | basic |
| BBa_K3308000 | [IMPDH-1 N (1-55)] | BBa_K3308044 | basic |
| BBa_K3308000 | [Npu-PCC73102 DnaE N]+stop | BBa_K3308045 | basic |
| BBa_K3308001 | [IMPDH-1 N (56-101)] + stop | BBa_K3308047 | basic |
| BBa_K3308001 | [Npu-PCC73102 DnaE C]- | BBa_K3308048 | basic |
| BBa_K3308002 | [IMPDH-1 C (1-35)]- | BBa_K3308051 | basic |
| BBa_K3308002 | [Tvo VMA C48 N] + stop | BBa_K3308052 | basic |
| BBa_K3308003 | [Tvo VMA C48 C] | BBa_K3308053 | basic |
| BBa_K3308003 | -[IMPDH-1 C (36-41)] | BBa_K3308054 | basic |
| BBa_K3308003 | -SICST-GB1+ stop | BBa_K3308055 | basic |
| BBa_K3308004 | gpD-GIGGG | BBa_K3308056 | basic |
| BBa_K3308004 | [IMPDH-1 N (1-55)]-[IMPDH-1 N (56-101) + stop | BBa_K3308057 | basic |
| BBa_K3308005 | [IMPDH-1 C (1-35)]-[IMPDH-1 C (36-41)] | BBa_K3308058 | basic |
| BBa_K3308005 | -SICST-GB1 + stop | BBa_K3308059 | basic |
| BBa_K3308006 | gpD-GIGGG-SICST-GB1+ stop | BBa_K3308060 | basic |
| BBa_K3308007, BBa_K3308081 | GB1-GTNPC- | BBa_K3308065 | basic |
| BBa_K3308007, BBa_K3308081 | [NrdJ-1 N (1-4)]- | BBa_K3308066 | basic |
| BBa_K3308007, BBa_K3308081 | [gp41-1 N] + stop | BBa_K3308067 | basic |
| BBa_K3308008, BBa_K3308082 | [gp41-1 C]- | BBa_K3308068 | basic |
| BBa_K3308008, BBa_K3308082 | [NrdJ-1 N (5-104)] + stop | BBa_K3308069 | basic |
| BBa_K3308009, BBa_K3308083 | [NrdJ-1 C (1-4)]- | BBa_K3308070 | basic |
| BBa_K3308009, BBa_K3308083 | [Tvo VMA C48 N] + stop | BBa_K3308071 | basic |
| BBa_K3308010, BBa_K3308084 | [Tvo VMA C48 C]- | BBa_K3308072 | basic |
| BBa_K3308010, BBa_K3308084 | [NrdJ-1 C (5-41)]- | BBa_K3308073 | basic |
| BBa_K3308010, BBa_K3308084 | SEIVL-gpD + stop | BBa_K3308074 | basic |
| BBa_K3308011, BBa_K3308085 | [NrdJ-1 C (1-4)]-[NrdJ-1 C (5-41)]- | BBa_K3308075 | basic |
| BBa_K3308011, BBa_K3308085 | SEIVL-gpD+ stop | BBa_K3308076 | basic |
| BBa_K3308012, BBa_K3308086 | GB1-GTNPC-SEIVL-gpD + stop | BBa_K3308077 | basic |
| BBa_K3308012, BBa_K3308086 | GB1-GTNPC- | BBa_K3308078 | basic |
| BBa_K3308013, BBa_K3308087 | [NrdJ-1 N (1-4)]-[NrdJ-1 N (5-104)] + stop | BBa_K3308079 | basic |
| BBa_K3308115, BBa_K3308119 | Lumio- | BBa_K3308121 | basic |
| BBa_K3308115, BBa_K3308119 | aey | BBa_K3308122 | basic |
| BBa_K3308115 | [Npu-PCC73102 DnaE N (1-28)] | BBa_K3308123 | basic |
| BBa_K3308115 | [Tvo VMA C48 N]+ stop | BBa_K3308124 | basic |
| BBa_K3308116 | [Tvo VMA C48 C] | BBa_K3308125 | basic |
| BBa_K3308116 | [Npu-PCC73102 DnaE N (29-102)] | BBa_K3308126 | basic |
| BBa_K3308117 | [Npu-PCC73102 DnaE C (1-19)] | BBa_K3308127 | basic |
| BBa_K3308117 | [NrdJ-1 N]+ stop | BBa_K3308128 | basic |
| BBa_K3308118 | [NrdJ-1 C]- | BBa_K3308129 | basic |
| BBa_K3308118 | [Npu-PCC73102 DnaE C (20-36)] | BBa_K3308130 | basic |
| BBa_K3308118, BBa_K3308120 | CFN- | BBa_K3308131 | basic |
| BBa_K3308118, BBa_K3308120 | GB1 + stop | BBa_K3308132 | basic |
| BBa_K3308119 | [Npu-PCC73102 DnaE N (1-28)]-[Npu-PCC73102 DnaE N (29-102)] + stop | BBa_K3308133 | basic |
| BBa_K3308120 | [Npu-PCC73102 DnaE C (1-19)]-[Npu-PCC73102 DnaE C (20-36)] | BBa_K3308134 | basic |
| BBa_K3308027, BBa_K3308028 | gp41-1-C | BBa_K3308095 | basic |
| BBa_K3308027, BBa_K3308028 | gp41-1-CF | BBa_K3308096 | basic |
| All split linker composite parts | GS Linker | BBa_K3308097 | basic |
| BBa_K3308027, BBa_K3308030, BBa_K3308032 | gp41-8-NF | BBa_K3308098 | basic |
| BBa_K3308027, BBa_K3308030, BBa_K3308032 | gp41-8-N + stop | BBa_K3308099 | basic |
| BBa_K3308028 | NrdJ-1 NF | BBa_K3308100 | basic |
| BBa_K3308028 | NrdJ-1 N + stop | BBa_K3308101 | basic |
| BBa_K3308029, BBa_K3308034 | gp41-8 C | BBa_K3308102 | basic |
| BBa_K3308029, BBa_K3308034 | gp41-8 CF | BBa_K3308103 | basic |
| BBa_K3308029, BBa_K3308031 | gp41-1 NF | BBa_K3308104 | basic |
| BBa_K3308029, BBa_K3308031 | gp41-1 N + stop | BBa_K3308105 | basic |
| BBa_K3308030 | NrdJ-1 C | BBa_K3308106 | basic |
| BBa_K3308030 | NrdJ-1 CF | BBa_K3308107 | basic |
| BBa_K3308031, BBa_K33080312, BBa_K3308035, BBa_K3308036 | Sce-VMA NF | BBa_K3308108 | basic |
| BBa_K3308031, BBa_K33080312, BBa_K3308035, BBa_K3308036 | Sce-VMA N | BBa_K3308109 | basic |
| BBa_K3308033, BBa_K3308034, BBa_K3308035, BBa_K3308036 | Sce-VMA C | BBa_K3308110 | basic |
| BBa_K3308033, BBa_K3308034, BBa_K3308035, BBa_K3308036 | Sce-VMA CF | BBa_K3308111 | basic |
| BBa_K3308033, BBa_K3308034, BBa_K3308035, BBa_K3308036 | GB1 + stop | BBa_K3308112 | basic |
| BBa_K3308035 | linker a - GS- ANF-ACF- GS | BBa_K3308113 | basic |
| BBa_K3308036 | linker ab - GS-ANF-ACF-GS-BNF-BCF-GS | BBa_K3308114 | basic |
Composite parts
Nested inteins set 1:
| Construct | Classification | Part Number | Type |
| pVS00028 | N1 | BBa_K3308007 | composite |
| pVS00029 | N2 | BBa_K3308008 | composite |
| pVS00030 | C1 | BBa_K3308009 | composite |
| pVS00031 | C2 | BBa_K3308010 | composite |
| pVS00043 | NSP | BBa_K3308011 | composite |
| pVS00033 | CSP | BBa_K3308012 | composite |
| pVS00034 | MSP | BBa_K3308013 | composite |
Nested inteins set 2:
| Construct | Classification | Part Number | Type |
| pVS00020 | N1 | BBa_K3308000 | composite |
| pVS00021 | N2 | BBa_K3308001 | composite |
| pVS00022 | C1 | BBa_K3308002 | composite |
| pVS00023 | C2 | BBa_K3308003 | composite |
| pVS00042 | NSP | BBa_K3308004 | composite |
| pVS00025 | CSP | BBa_K3308005 | composite |
| pVS00026 | MSP | BBa_K3308006 | composite |
Nested inteins set 3:
| Construct | Classification | Part Number | Type |
| pVS000036 | N1 | BBa_K3308115 | composite |
| pVS000037 | N2 | BBa_K3308116 | composite |
| pVS000038 | C1 | BBa_K3308117 | composite |
| pVS000039 | C2 | BBa_K3308118 | composite |
| pVS000040 | NSP | BBa_K3308119 | composite |
| pVS000041 | CSP | BBa_K3308120 | composite |
Split linker constructs:
| Construct | Classification | Part Number | Type |
| pHG000044 | linker-AB | BBa_K3308027 | composite |
| pHG000045 | linker-AC | BBa_K3308028 | composite |
| pHG000047 | linker-BA | BBa_K3308029 | composite |
| pHG000052 | linker-CB | BBa_K3308030 | composite |
| pHG000059 | N-VMA-A | BBa_K3308031 | composite |
| pHG000060 | N-VMA-B | BBa_K3308032 | composite |
| pHG000063 | C-VMA-A | BBa_K3308033 | composite |
| pHG000064 | C-VMA-B | BBa_K3308034 | composite |
| pHG000067 | cis-A | BBa_K3308035 | composite |
| pHG000068 | cis-AB | BBa_K3308036 | composite |
Basic parts
Basic parts: