Team:Paris/Freezer
From 2009.igem.org
Contents |
Freezer
- Primers
- PCR products
- Plasmids: MiniPrep Products
- Digestion Products
- Unexpected digestion results
- Strains (Glycerol stock)
- Color code for sticker
- Vector
- Insert
- PCR product
- Plasmid
- Kanamycin
- Ampicilin
- Chloranphenicol
Primers
[O1] Rev P1 | |||||
---|---|---|---|---|---|
Description | tolR | ||||
Sequence (5' > 3') | TCTGCTGCAGCGGCCGCTACTAGTATTAGTAAGGCACATCTTTTGCGCCACCG | ||||
Designer | D. Bikard | ||||
Worked |
[O2] Fwd P2 | |||||
---|---|---|---|---|---|
Description | tolR (DO NOT USE ERROR) | ||||
Sequence (5' > 3') | TCTGGAATTCGCGGCCGCTTCTAGATGAAATCAACATTGTACCGTTGCTGGACGTACTG | ||||
Designer | D. Bikard | ||||
Worked |
[O3] Rev P3 | |||||
---|---|---|---|---|---|
Description | tolA | ||||
Sequence (5' > 3') | TCTGCTGCAGCGGCCGCTACTAGTATTATTACGGTTTGAAGTCCAATGGCG | ||||
Designer | D. Bikard | ||||
Worked |
[O4] Rev P4 | |||||
---|---|---|---|---|---|
Description | tolA (ERROR: Not designed for N-term fusion) | ||||
Sequence (5' > 3') | TCTGGAATTCGCGGCCGCTTCTAGAATGGGTAATACTAAAAACAATGGCGC | ||||
Designer | D. Bikard | ||||
Worked |
[O5] Rev P5 | |||||
---|---|---|---|---|---|
Description | tatABC | ||||
Sequence (5' > 3') | TCTGCTGCAGCGGCCGCTACTAGTATTATTATTCTTCAGTTTTTTCGCTTTCTGC | ||||
Designer | D. Bikard | ||||
Worked |
[06] Fwd P6 | |||||
---|---|---|---|---|---|
Description | tatABC (without RBS) | ||||
Sequence (5' > 3') | TCTGGAATTCGCGGCCGCTTCTAGAATGGGTGGTATCAGTATTTGG | ||||
Designer | D. Bikard | ||||
Worked |
[O7] Rev P7 | |||||
---|---|---|---|---|---|
Description | clyA fusion | ||||
Sequence (5' > 3') | TTTCTGCAGCGGCCGCTACTAGTGACTTCAGGTACCTCAAAGAGTGTC | ||||
Designer | D. Bikard | ||||
Worked |
[O8] Fwd P8 | |||||
---|---|---|---|---|---|
Description | clyA fusion (ERROR: No poly-Gly linker) | ||||
Sequence (5' > 3') | TTTGAATTCGCGGCCGCTTCTAGAATGACTGAAATCGTTGCAGATAAAA | ||||
Designer | D. Bikard | ||||
Worked |
[O9] clyA_Rw | |||||
---|---|---|---|---|---|
Description | clyA | ||||
Sequence (5' > 3') | GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTAGACTTCAGGTACCTCAAAGAG | ||||
Designer | G. Cambray | ||||
Not Tested |
[O10] clyA_fusion_polyG_Rw | |||||
---|---|---|---|---|---|
Description | clyA fusion (with 5-Gly linker) | ||||
Sequence (5' > 3') | GTTTCTTCCTGCAGCGGCCGCTACTAGTGCCGCCGCCGCCGCCGACTTCAGGTACCTCAAAGAG | ||||
Designer | G. Cambray | ||||
Not Tested |
[O11] ompA_signal_Fw | |||||
---|---|---|---|---|---|
Description | ompA signal for export to periplasm - from expression vector RBS as in pIN-III-ompA-2 vector - keeps vector original RBS | ||||
Sequence (5' > 3') | GTTTCTTCGAATTCGCGGCCGCTTCTAGAGAACGAGGGCAAAAAATGAAAAAG | ||||
Designer | G. Cambray | ||||
Not Tested |
[O12] ompA_signal_fusion_Rw | |||||
---|---|---|---|---|---|
Description | ompA signal for export to periplasm - from expression vector pIN-III-ompA-2 - original RBS from the vector | ||||
Sequence (5' > 3') | GTTTCTTCCTGCAGCGGCCGCTACTAGTGGCCTGCGCTACGGTAGCGAAAC | ||||
Designer | G. Cambray | ||||
Not Tested |
[O13] tolR_II_Fusion_Fw | |||||
---|---|---|---|---|---|
Description | For fusion of tolR domain II (standard 23) | ||||
Sequence (5' > 3') | GTTTCTTCGAATTCGCGGCCGCTTCTAGAGAGGTCGATCTGCCAGACGCTAC | ||||
Designer | G. Cambray | ||||
Not Tested |
[O14] tatABC_RBS_Fw | |||||
---|---|---|---|---|---|
Description | Amplification from tatA - contains RBS | ||||
Sequence (5' > 3') | TTGAATTCGCGGCCGCTTCTAGAAGGAGGTATATAATGATGGGTGGTATCAGTATTTGG | ||||
Designer | G. Cambray | ||||
Not Tested |
[O15] TE3_fusion_Fw | |||||
---|---|---|---|---|---|
Description | Fusion of N-term domain of colicin E3 (standard 23) | ||||
Sequence (5' > 3') | GTTTCTTCGAATTCGCGGCCGCTTCTAGAAGCGGTGGCGATGGACGCGGCCATAAC | ||||
Designer | G. Cambray | ||||
Not Tested |
[O16] TE3_Rw | |||||
---|---|---|---|---|---|
Description | Amplification of N-term domain of colicin E3 (standard 23) | ||||
Sequence (5' > 3') | GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTAGCGTTCATAATTTCGCTCAG | ||||
Designer | G. Cambray | ||||
Not Tested |
[O17] Tg3p_Rw | |||||
---|---|---|---|---|---|
Description | Amplification of N-term domain of M13's g3p | ||||
Sequence (5' > 3') | GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTAACACTGAGTTTCGTCACCAGTAC | ||||
Designer | G. Cambray | ||||
Not Tested |
[O18] Tg3p_fusion_Fw | |||||
---|---|---|---|---|---|
Description | Fusion of N-term domain of M13's g3p (standard 23) | ||||
Sequence (5' > 3') | GTTTCTTCGAATTCGCGGCCGCTTCTAGAAAAAAATTATTATTCGCAATTCCTTTAG | ||||
Designer | G. Cambray | ||||
Not Tested |
[O19] g3p_F | |||||
---|---|---|---|---|---|
Description | g3p | ||||
Sequence (5' > 3') | GTTTCTTCGAATTCGCGGCCGCTTCTAGAGCTCCGCTGAAACTGTTGAAAGTTG | ||||
Designer | G. Cambray | ||||
Not Tested |
[O20] g3p_R | |||||
---|---|---|---|---|---|
Description | g3p | ||||
Sequence (5' > 3') | GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTAAGACTCCTTATTACGCAG | ||||
Designer | G. Cambray | ||||
Not Tested |
[O21] Ompa_linker_F | |||||
---|---|---|---|---|---|
Description | For standardisation of OmpA linker (BBaK103006) | ||||
Sequence (5' > 3') | TTGAATTCGCGGCCGCTTCTAGAAGGAGGTATATAATGAAAGCTACTAAACTGGTACTGG | ||||
Designer | G. Cambray | ||||
Not Tested |
[O22] Ompa_linker_R | |||||
---|---|---|---|---|---|
Description | For standardisation of OmpA linker (BBaK103006) | ||||
Sequence (5' > 3') | GTTTCTTCCTGCAGCGGCCGCTACTAGTAGAGCTCCCTCCTCCAGAACC | ||||
Designer | G. Cambray | ||||
Not Tested |
[O23] OmpAsignal-TE3_seq_Fw | |||||
---|---|---|---|---|---|
Description | ??? | ||||
Sequence (5' > 3') | CACAAATAGCGAAAGATGAC | ||||
Designer | G. Cambray | ||||
Not Tested |
[O24] OmpAsignal-TE3_seq_Rw | |||||
---|---|---|---|---|---|
Description | ??? | ||||
Sequence (5' > 3') | ATCATCTGCGGGTAATGATG | ||||
Designer | G. Cambray | ||||
Not Tested |
[O25] VAMP2_Seq_Fw | |||||
---|---|---|---|---|---|
Description | ??? | ||||
Sequence (5' > 3') | ACTCACTATAGGGAGACCACA | ||||
Designer | G. Cambray | ||||
Not Tested |
[O26] VAMP2_Seq_Rv | |||||
---|---|---|---|---|---|
Description | ??? | ||||
Sequence (5' > 3') | GGGCTTTGTTAGCAGCCGGAT | ||||
Designer | G. Cambray | ||||
Not Tested |
[O27] ClyA_seq_Fw | |||||
---|---|---|---|---|---|
Description | ??? | ||||
Sequence (5' > 3') | GCAGCTATTTCCAGTCACAG | ||||
Designer | G. Cambray | ||||
Not Tested |
[O28] ClyA_seq_Rw | |||||
---|---|---|---|---|---|
Description | ??? | ||||
Sequence (5' > 3') | CGCCCGCAGCAATAGAATAG | ||||
Designer | G. Cambray | ||||
Not Tested |
[O29] PFecA_Fw | |||||
---|---|---|---|---|---|
Description | ??? | ||||
Sequence (5' > 3') | GTTTCTTCGAATTCGCGGCCGCTTCTAGAGTACGCGGTACTGGATAAACATTTC | ||||
Designer | G. Cambray | ||||
Not Tested |
[O30] PFecA_Rv | |||||
---|---|---|---|---|---|
Description | ??? | ||||
Sequence (5' > 3') | GTTTCTTCCTGCAGCGGCCGCTACTAGTATTTTTGTTGTGTTCAGCTATGAGT | ||||
Designer | G. Cambray | ||||
Not Tested |
PCR products
PCR products | ||||||||||
Number | Name | Oligo F | Oligo R | Tm | Matrix | Length | Comments | Quantity | []ng/µL | Date |
A1 | g3p_std | [O19] g3p_F | [O20] g3p_R | Tm | Matrix | Length | For standardisation of g3p and adding SacI site | ? | ? ng/µL | ? |
A2 | OmpA_L_std | [O21] Ompa_linker_F | [O22] Ompa_linker_R | Tm | Matrix | Length | For standardisation of OmpA linker (BBaK103006) | ? | ? ng/µL | ? |
http://parts.mit.edu/r/parts/images/partbypart/icon_terminator.png
http://parts.mit.edu/images/partbypart/icon_regulatory.png
http://parts.mit.edu/r/parts/images/partbypart/icon_rbs.png
http://parts.mit.edu/r/parts/images/partbypart/icon_reporter.png
http://parts.mit.edu/registry/images/7/7b/Part_icon_cds.png
http://parts.mit.edu/registry/images/9/9f/Part_icon_dna.png
http://parts.mit.edu/registry/images/2/2e/Part_icon_generator.png
http://parts.mit.edu/registry/images/a/ab/Part_icon_plasmid.png
Plasmids: MiniPrep Products
Plasmids: MiniPrep Products | ||||
Number | Name | Description | Products in use | Date |
P1 | pSB2K3 | General plasmid for cloning step after PCR | Products in use | 08/03 |
P2 | Bba-J61002 | Plasmid backbone, it allows us to change promoter RFP as promoter | Products in use | 08/03 |
P3 | Bba-J32015 | PelB leader sequence | Products in use | 08/03 |
P4 | Bba-B0014 | Double terminator bidirectionnal (B0012-B0011) | Products in use | 08/03 |
P5 | Bba-I14032 | pLac | Products in use | 08/03 |
P6 | BBa_K103006 | OmpA linker (464bp) | Products in use | 08/03 |
P7 | Bba-R0040 | TetR repressible promoter | Products in use | 08/03 |
P8 | Bba-I0500 | pBad/araC | Products in use | 08/03 |
P9 | ΔN-Syn1A | ??? | Products in use | 08/03 |
P10 | His6-SNAP25 | ??? | Products in use | 08/03 |
P11 | VAMP2-HiB6 | ??? | Products in use | 08/03 |
P12 | g3p | g3p in pHEN2 plasmid | Products in use | 08/04 |
Digestion Products
Red = Vector
Yellow = Insert
Digestion Products | |||||||||
---|---|---|---|---|---|---|---|---|---|
Number | Product Name | Matrix Name | Enz. 1 | Enz. 2 | Size | Description | Product Left | []ng/µL | Date |
D1 | P3 | BBa-J32015 | XbaI | PstI | Size | ??? | ??? | ??? | 08/04 |
D2 | P4 | BBa-B0014 | XbaI | PstI | Size | ??? | ??? | ??? | 08/04 |
D3 | P5 | BBa-I14032 | XbaI | PstI | Size | ??? | ??? | ??? | 08/04 |
D4 | P7 | Bba-R0040 | XbaI | PstI | Size | ??? | ??? | ??? | 08/04 |
D5 | P8 | BBa-10500 | XbaI | PstI | Size | ??? | ??? | ??? | 08/04 |
D6 | ??? | ??? | XbaI | PstI | Size | ??? | ??? | ??? | 08/04 |
D7 | ??? | ??? | XbaI | PstI | Size | ??? | ??? | ??? | 08/04 |
Unexpected digestion results
Digestion Products | ||||||
---|---|---|---|---|---|---|
Number | Matrix Name | Enz. 1 | Enz. 2 | Expected Size | Description | Date |
Strains (Glycerol stock)
Strains glycerol stock | ||||
---|---|---|---|---|
Number | R | Name | Description | Date |
07/20 | ||||
S2 | K | K12 MG 1655 ΔrecA::Kan | Standard E.Coli strain, Kan resistance | 07/20 |
S3 | K | W3110 Δnlp1 | From Daniel Rodriguez, Kan resistance | 07/23 |
TOP10 | TOP10 | Competent bacteria | 07/27 | |
S4 | TB28 | No kanamycin resistance | 07/29 | |
S5 | K | MG4 | Strain needs to be grown on media with [NaCl]>=0.5%, preferably 1%NaCl | 07/29 |
S6 | DH5α | DH5α E. Coli strain for chimically competent (Pasteur) | 07/30 | |
S7 | A | DH5α(g3p) | DH5α E. Coli strain with g3p in plasmid, Amp resistance | 07/30 |
S8 | K | DH5α(pSB2K3) | DH5α E. Coli strain with pSB2K3 plasmid, Kan resistance | 08/03 |
S9 | A | DH5α(BBa_J61002) | DH5α E. Coli strain with BBa_J61002 in plasmid, Amp resistance | 08/03 |
S10 | K | DH5α(BBa_J32015) | DH5α E. Coli strain with BBa_J32015 in plasmid, Kan resistance | 08/03 |
S11 | K | DH5α(BBa_B0014) | DH5α E. Coli strain with BBa_B0014 in plasmid, Kan resistance | 08/03 |
S12 | K | DH5α(BBa_I14032) | DH5α E. Coli strain with BBa_I14032 in plasmid, Kan resistance | 08/03 |
S13 | K | DH5α(BBa_K103006) | DH5α E. Coli strain with BBa_K103006 in plasmid, Kan resistance | 08/03 |
S14 | A | DH5α(BBa_R0040) | DH5α E. Coli strain with BBa_R0040 in plasmid, Amp resistance | 08/03 |
S15 | K | DH5α(BBa_10500) | DH5α E. Coli strain with BBa_10500 in plasmid, Kan resistance | 08/03 |
S16 | K | DH5α(ΔN-Syn1A) | DH5α E. Coli strain with ΔN-Syn1A in plasmid, Kan resistance | 08/03 |
S17 | A | DH5α(His6-SNAP25) | DH5α E. Coli strain with His6-SNAP25 in plasmid, Amp resistance | 08/03 |
S18 | A | DH5α(Vamp2-His6) | DH5α E. Coli strain with Vamp2-His6 in plasmid, Amp resistance | 08/03 |
S19 | A | DH5α(puc19) | DH5α E. Coli strain with puc19 plasmid, Amp resistance | 07/30 |