Team:Paris/Freezer

From 2009.igem.org

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|width=15%| Products in use

Revision as of 08:02, 4 August 2009

Contents

Freezer


  • 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 Matrix Length Comments Product Left ? []ng/µL Date

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

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  ???  ??? XbaI PstI Size  ???  ???  ??? 08/04
D2  ???  ??? XbaI PstI Size  ???  ???  ??? 08/04
D3  ???  ??? XbaI PstI Size  ???  ???  ??? 08/04
D4  ???  ??? XbaI PstI Size  ???  ???  ??? 08/04
D5  ???  ??? 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
S1 DH5α DH5α E. Coli strain for chimically competent 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

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