Team:UNIPV-Pavia/Freezer

From 2010.igem.org

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<font class="BBok">BBa_J23118><br/>_4C5</font
+
<font class="BBok">BBa_J23118><br/>_4C5</font>
<br/>
<br/>
(it was <partinfo>BBa_J23118</partinfo><br/>_4C5-1)
(it was <partinfo>BBa_J23118</partinfo><br/>_4C5-1)

Revision as of 13:52, 30 August 2010

FREEZER MANAGEMENT



Biobrick name Description Creation date Strain Vector Quality control Length Store

<partinfo>BBa_E2050</partinfo>

mOrange

08/06/2010

E. coli DH5-alpha

<partinfo>pSB2K3</partinfo>

Seq ok (MIT); orange pellet when contained in <partinfo>pSB2K3</partinfo>

744 bpBioBricks 2010

<partinfo>BBa_K165018</partinfo>

NLS-ADH1 terminator (Silver Standard 5' and 3')

08/06/2010

E. coli DH5-alpha

<partinfo>J63009</partinfo> (Amp)

Seq partially confirmed (MIT) because it lacks a T at the 3', but the ADH1 terminator (i.e. <partinfo>BBa_K165010</partinfo>) is ok. The correct ApE file is here.

253 bpBioBricks 2010

<partinfo>BBa_K165037</partinfo>

tef2 promoter

08/06/2010

E. coli DH5-alpha

<partinfo>pSB1AK3</partinfo>

Seq ok (MIT).

403 bpBioBricks 2010

<partinfo>BBa_P1004</partinfo>

Chloramphenicol resistence
cassette

08/06/2010

E. coli DH5-alpha

<partinfo>pSB1A1</partinfo>

Seq ok (MIT 2009); length ok.

769 bpBioBricks 2010

<partinfo>BBa_J61001</partinfo>

R6K origin

08/06/2010

E. coli DH5-alpha

<partinfo>pSB1A2</partinfo>

Seq ok (MIT).

406 bpBioBricks 2010

<partinfo>BBa_K081008</partinfo>

RBS-luxI

08/06/2010

E. coli DH5-alpha

<partinfo>pSB1A2</partinfo>

Seq ok (MIT); length ok.

664 bpBioBricks 2010

<partinfo>BBa_K125500</partinfo>

GFP fusion brick (Silver Standard 5', Knight Standard 3')

08/06/2010

E. coli DH5-alpha

<partinfo>pSB1A2</partinfo>

Seq partially confirmed (MIT) for unknown reasons: the alignment is fully correct. The ApE file with the correct prefix and suffix is here. Length ok.

718 bpBioBricks 2010

<partinfo>BBa_K208001</partinfo>
(stab-1)

phasin phaP
(Silver Standard 5' and 3')

24/06/2010

E. coli ???

<partinfo>pSB3K3</partinfo>

Seq ok (BMR); length ok.

579 bpBioBricks 2010

<partinfo>BBa_K208001</partinfo>
(stab-2)

phasin phaP
(Silver Standard 5' and 3')

24/06/2010

E. coli ???

<partinfo>pSB3K3</partinfo>

Seq ok (BMR); length ok.

579 bpBioBricks 2010

<partinfo>BBa_J72007</partinfo>

BamHI methyltransferase encoding CRIM plasmid

01/07/2010

E. coli ???

HC Cm resistance

No plasmid in the clone.
For more informations click here.

BioBricks 2010

<partinfo>BBa_J72008</partinfo>

phi80 integration helper plasmid pInt80-649

01/07/2010

E. coli ???

Plasmid (Amp 50 ug/ml resistance)

Length ok.

6335 bpBioBricks 2010

<partinfo>BBa_J72013</partinfo>

BglII methyltransferase encoding CRIM plasmid

01/07/2010

E. coli ???

HC Cm resistance

No plasmid in the clone.
For more informations click here.

BioBricks 2010

<partinfo>BBa_K105012</partinfo>

10 aa flexible protein domain linker

06/08/2010

E. coli DH5-alpha

<partinfo>pSB1A2</partinfo>

-

30 bp BioBricks 2010

<partinfo>BBa_J13002</partinfo>

p(tetR)+RBS34

16/08/2010

E. coli TOP10

<partinfo>pSB1A2</partinfo>

-

74 bp BioBricks 2010

<partinfo>BBa_I13522</partinfo>

Untagged GFP behind a constitutive promoter

16/08/2010

E. coli TOP10

<partinfo>pSB1A2</partinfo>

-

937 bp BioBricks 2010
Ligation name Description Creation date Strain Vector Quality control Length Store

I0
(it was I0-2)

<partinfo>BBa_K125500</partinfo> (E-S)
+
<partinfo>BBa_B0015</partinfo> (E-X)

11/06/2010

E. coli DH5-alpha

<partinfo>pSB1AK3</partinfo>

Seq ok (BMR); length ok.

853 bpLigations 2010

I1
(it was I1-2)

<partinfo>BBa_E2050</partinfo> (E-S)
+
<partinfo>BBa_K165018</partinfo> (E-X)

11/06/2010

E. coli DH5-alpha

<partinfo>J63009</partinfo> (Amp)

Seq ok (BMR), with an additional barcode (25bp) after the stop codon of mOrange; length ok.

1029 bp Ligations 2010

I2
(it was I2-1)

<partinfo>BBa_P1004</partinfo> (E-S)
+
<partinfo>BBa_B0015</partinfo> (E-X)

11/06/2010

E. coli DH5-alpha

<partinfo>pSB1AK3</partinfo>

Seq ok (BMR); Cm resistance ok; length ok.

906 bpLigations 2010

I3
(it was I3-1)

<partinfo>BBa_K081008</partinfo> (E-S)
+
<partinfo>BBa_B0015</partinfo> (E-X)

11/06/2010

E. coli DH5-alpha

<partinfo>pSB1AK3</partinfo>

Seq TWO PLASMIDS IN THIS CLONE!(BMR), two overlapped traces; length ok.

801 bpLigations 2010

I4
(it was I4-2)

<partinfo>BBa_K165037</partinfo> (S-P)
+
I1 (X-P)

18/06/2010

E. coli DH5-alpha

<partinfo>pSB1AK3</partinfo>

Seq ok (BMR); length ok.

1439 bp Ligations 2010

I5
(it was I5-1)

I2 (E-S)
+
<partinfo>BBa_J61001</partinfo> (E-X)

18/06/2010

E. coli TOP10

<partinfo>pSB1A2</partinfo>

Seq ok (BMR); Cm resistance ok; length ok.

1320 bpLigations 2010

I6
(it was I6-2)

I3 (E-S)
+
<partinfo>BBa_T9002</partinfo> (E-X)

18/06/2010

E. coli DH5-alpha

<partinfo>pSB1A3</partinfo>

Seq LONG PART(BMR), but the ligation worked and the sequenced portions are ok; length ok.

2754 bp Ligations 2010

I7
(it was I7-3)

<partinfo>BBa_J23118</partinfo> (E-S)
+
I6 (X-P)

25/06/2010

E. coli TOP10

<partinfo>BBa_J61002</partinfo>
(Amp)

Length ok.

2795 bp Ligations 2010

I8
(it was I8-5)

<partinfo>BBa_J23110</partinfo> (E-S)
+
I6 (X-P)

25/06/2010

E. coli TOP10

<partinfo>BBa_J61002</partinfo>
(Amp)

Length ok.

2795 bp Ligations 2010

I9
(it was I9-1)

<partinfo>BBa_J23111</partinfo> (E-S)
+
I6 (X-P)

25/06/2010

E. coli TOP10

<partinfo>BBa_J61002</partinfo>
(Amp)

Length ok.

2795 bp Ligations 2010

I10
(it was I10-1)

<partinfo>BBa_J23116</partinfo> (E-S)
+
I6 (X-P)

25/06/2010

E. coli TOP10

<partinfo>BBa_J61002</partinfo>
(Amp)

Length ok.

2795 bp Ligations 2010

I12
(it was I12-2)

<partinfo>BBa_J23105</partinfo> (S-P)
+
I6 (X-P)

16/07/2010

E. coli TOP10

<partinfo>BBa_J61002</partinfo>
(Amp)

Seq ok (BMR), length ok.

2795 bp Ligations 2010

I14
(it was I14-1)

<partinfo>BBa_J23106</partinfo> (S-P)
+
I3 (X-P)

16/07/2010

E. coli DH5-alpha

<partinfo>BBa_J61002</partinfo>
(Amp)

Seq ok (BMR), length ok.

842 bp Ligations 2010

RING

I5 (X-S) on itself:
Cm resistance (<partinfo>BBa_P1004</partinfo>) + TT (<partinfo>BBa_B0015</partinfo>) + R6K ori (<partinfo>BBa_J61001</partinfo>)

16/07/2010

E. coli DH5-alpha

Plasmid (Cm resistance, conditional thermosensitive ORI; works in PIR strains)

Tested here.

1328 bp

Ligations 2010

I15
(it was I15-1)

<partinfo>BBa_J23110</partinfo> (S-P)
+
I3 (X-P)

23/07/2010

E. coli DH5-alpha

<partinfo>BBa_J61002</partinfo>
(Amp)

Length ok.

842 bp Ligations 2010

I16
(it was I16-1)

<partinfo>BBa_J23116</partinfo> (S-P)
+
I3 (X-P)

16/07/2010

E. coli DH5-alpha

<partinfo>BBa_J61002</partinfo>
(Amp)

Seq ok (BMR), length ok.

842 bp Ligations 2010

I17
(it was I17-1)

<partinfo>BBa_J23101</partinfo> (S-P)
+
I3 (X-P)

16/07/2010

E. coli DH5-alpha

<partinfo>BBa_J61002</partinfo>
(Amp)

Seq ok (BMR), length ok.

842 bp Ligations 2010

I18
(it was I18-1)

<partinfo>BBa_J23105</partinfo> (S-P)
+
I3 (X-P)

16/07/2010

E. coli DH5-alpha

<partinfo>BBa_J61002</partinfo>
(Amp)

Seq ok (BMR), length ok.

842 bp Ligations 2010

I19
(it was I19-1)

<partinfo>BBa_J23106</partinfo> (S-P)
+
I3 (X-P)

16/07/2010

E. coli DH5-alpha

<partinfo>BBa_J61002</partinfo>
(Amp)

Seq ok (BMR), length ok.

842 bp Ligations 2010

I20
(it was I20M-1)

Pha-10S-1 (X-S) (infos here)
+
<partinfo>pSB1AK3</partinfo> (X-S)

13/08/2010

E. coli DH5-alpha

<partinfo>pSB1AK3</partinfo>

Length ok.

576 bp

Ligations 2010

I21
(it was I21-4)

Pha-SS-1 (X-S) (infos here)
+
<partinfo>pSB1A3</partinfo> (X-S)

10/08/2010

E. coli DH5-alpha

<partinfo>pSB1A3</partinfo>

Length ok.

576 bp

Ligations 2010

I22
(it was I22-1)

<partinfo>BBa_K081009</partinfo> (E-S)
+
<partinfo>BBa_B0015</partinfo> (E-X)

10/08/2010

E. coli DH5-alpha

<partinfo>pSB1AK3</partinfo>

Length ok.

825 bp

Ligations 2010

I23
(it was I23-1)

<partinfo>BBa_R0062</partinfo> (S-P)
+
I3 (X-P)

10/08/2010

E. coli DH5-alpha

<partinfo>pSB1A2</partinfo>

Length ok.

725 bp

Ligations 2010

I24
(it was I24-1)

<partinfo>BBa_R0062</partinfo> (S-P)
+
I22 (X-P)

13/08/2010

E. coli DH5-alpha

<partinfo>pSB1A2</partinfo>

Length ok.

886 bp

Ligations 2010

I26
(it was I26-2)

<partinfo>BBa_K105012</partinfo> (S-P)
+
I21-B (X-P)

13/08/2010

E. coli DH5-alpha

<partinfo>pSB1A2</partinfo>

Length ok.

612 bp

Ligations 2010

I29
(it was ??)

<partinfo>BBa_T9002</partinfo> (S-P)
+
I23 (X-P)

E. coli TOP10

<partinfo>pSB1A3</partinfo>

725 bp

Ligations 2010

I30
(it was ??)

<partinfo>BBa_T9002</partinfo> (S-P)
+
I24 (X-P)

E. coli TOP10

<partinfo>pSB1A3</partinfo>

886 bp

Ligations 2010

I31
(it was I31-1)

I20 (E-S)
+
<partinfo>BBa_K105012</partinfo> (E-X)

23/08/2010

E. coli DH5-alpha

<partinfo>pSB1A2</partinfo>

Length ok.

614 bp

Ligations 2010

I32
(it was I32-2)

I20 (E-S)
+
I21 (E-X)

23/08/2010

E. coli DH5-alpha

<partinfo>pSB1A3</partinfo>

Length ok.

1158 bp

Ligations 2010

I33
(it was I33-1)

I20 (E-S)
+
I0 (E-X)

23/08/2010

E. coli DH5-alpha

<partinfo>pSB1AK3</partinfo>

Length ok.

1437 bp

Ligations 2010

I34
(it was I34-1)

I21 (E-S)
+
I0 (E-X)

23/08/2010

E. coli DH5-alpha

<partinfo>pSB1AK3</partinfo>

Length ok.

1437 bp

Ligations 2010

I35
(it was I35-1)

I21 (E-S)
+
<partinfo>BBa_K105012</partinfo> (E-X)

23/08/2010

E. coli DH5-alpha

<partinfo>pSB1A2</partinfo>

Length ok.

614 bp

Ligations 2010

I36
(it was I36-7)

I21 (E-S)
+
I21 (E-X)

24/08/2010

E. coli DH5-alpha

<partinfo>pSB1A3</partinfo>

Length ok.

1158 bp

Ligations 2010

I37
(it was I37-1)

INTEIN (E-S)
+
I0 (E-X)

23/08/2010

E. coli DH5-alpha

<partinfo>pSB1AK3</partinfo>

Length ok.

1366 bp

Ligations 2010

I38
(it was I38-3)

<partinfo>BBa_F2620</partinfo> (E-S)
+
<partinfo>BBa_B0034</partinfo> (E-X)

23/08/2010

E. coli DH5-alpha

<partinfo>pSB1A2</partinfo>

Length ok.

1079 bp

Ligations 2010

I39
(it was I39-5)

<partinfo>BBa_J13002</partinfo> (S-P)
+
I20 (X-P)

24/08/2010

E. coli DH5-alpha

<partinfo>pSB1AK3</partinfo>

Length ok.

652 bp

Ligations 2010

I40
(it was I40-1)

I26 (E-S)
+
INTEIN (E-X)

25/08/2010

E. coli DH5-alpha

pMA (HC Amp resistance)

Length ok.

1125 bp

Ligations 2010

I41
(it was I41-1)

I26 (E-S)
+
I0 (E-X)

23/08/2010

E. coli DH5-alpha

<partinfo>pSB1AK3</partinfo>

Length ok.

1471 bp

Ligations 2010

I42
(it was ??)

CREAM (E-P)
+
<partinfo>BBa_K173001</partinfo> (E-P)

E. coli BW23474

CREAM

3068 bp

Ligations 2010

I43
(it was ??)

CREAM (E-P)
+
<partinfo>BBa_J23101</partinfo> (E-P)

E. coli BW23474

CREAM

3026 bp

Ligations 2010

I44
(it was ??)

CREAM (E-P)
+
<partinfo>BBa_I52002</partinfo> (E-P)

E. coli DB3.1

CREAM

3239 bp

Ligations 2010

I45
(it was ??)

CREAM (HindIII) on itself

E. coli BW23474

CREAM

2149 bp

Ligations 2010

I47
(it was ??)

<partinfo>BBa_J13002</partinfo> (S-P) + I33 (X-P)

E. coli DH5-alpha

<partinfo>pSB1A2</partinfo>

1517 bp

Ligations 2010

I48
(it was ??)

I39 (S-P) + I34 (X-P)

E. coli DH5-alpha

<partinfo>pSB1AK3</partinfo>

1233 bp

Ligations 2010

I49
(it was ??)

I39 (S-P) + I41 (X-P)

E. coli DH5-alpha

<partinfo>pSB1AK3</partinfo>

2129 bp

Ligations 2010

INTEIN

Intein

13/08/2010

E. coli TOP10

pMA
(HC Amp resistance)

Mr.Gene DNA synthesis

507 bp

Ligations 2010

CRIM
MR.G

CREAM fragment

13/08/2010

E. coli TOP10

pMK-RQ
(HC Kan resistance)

Mr.Gene DNA synthesis

829 bp

Ligations 2010

CREAM

CRIM
MR.G (A-A)
+
I5 (S-X)

21/08/2010

E. coli BW23474

CREAM

Length ok.

2149 bp

Ligations 2010

I7-4C5
(it was I7-4C5-2)

I7 (E-P)
+
<partinfo>pSB4C5</partinfo>(E-P)

16/07/2010

E. coli TOP10

<partinfo>pSB4C5</partinfo>

Seq ok (BMR), length ok.

2795 bp Ligations 2010

I8-4C5
(it was I8-4C5-2)

I8 (E-P)
+
<partinfo>pSB4C5</partinfo> (E-P)

16/07/2010

E. coli TOP10

<partinfo>pSB4C5</partinfo>

Seq ok (BMR), length ok.

2795 bp Ligations 2010

I10-4C5
(it was I10-4C5-1)

I10 (E-P)
+
<partinfo>pSB4C5</partinfo> (E-P)

16/07/2010

E. coli TOP10

<partinfo>pSB4C5</partinfo>

Length ok.

2795 bp

Ligations 2010

I12-4C5
(it was I12-4C5-1)

I12 (E-P)
+
<partinfo>pSB4C5</partinfo> (E-P)

25/07/2010

E. coli TOP10

<partinfo>pSB4C5</partinfo>

Length ok.

2795 bp

Ligations 2010

I14-4C5
(it was I14-4C5-2)

I14 (E-P)
+
<partinfo>pSB4C5</partinfo> (E-P)

23/07/2010

E. coli DH5-alpha

<partinfo>pSB4C5</partinfo>

Length ok.

842 bp

Ligations 2010

I15-4C5
(it was I15-4C5-2)

I15 (E-P)
+
<partinfo>pSB4C5</partinfo> (E-P)

13/08/2010

E. coli TOP10

<partinfo>pSB4C5</partinfo>

Length ok.

842 bp

Ligations 2010

I16-4C5
(it was I16-4C5-1)

I16 (E-P)
+
<partinfo>pSB4C5</partinfo> (E-P)

23/07/2010

E. coli DH5-alpha

<partinfo>pSB4C5</partinfo>

Length ok.

842 bp

Ligations 2010

I17-4C5
(it was I17-4C5-1)

I17 (E-P)
+
<partinfo>pSB4C5</partinfo> (E-P)

25/07/2010

E. coli DH5-alpha

<partinfo>pSB4C5</partinfo>

Length ok.

842 bp

Ligations 2010

I18-4C5
(it was I18-4C5-1)

I18 (E-P)
+
<partinfo>pSB4C5</partinfo> (E-P)

25/07/2010

E. coli DH5-alpha

<partinfo>pSB4C5</partinfo>

Length ok.

842 bp

Ligations 2010

I19-4C5
(it was I19-4C5-1)

I19 (E-P)
+
<partinfo>pSB4C5</partinfo> (E-P)

25/07/2010

E. coli DH5-alpha

<partinfo>pSB4C5</partinfo>

Length ok.

842 bp

Ligations 2010

I27
(it was ??)

I23 (E-P)
+
<partinfo>pSB4C5</partinfo> (E-P)

??

E. coli TOP10

<partinfo>pSB4C5</partinfo>

725 bp

Ligations 2010

I28
(it was ??)

I24 (E-P)
+
<partinfo>pSB4C5</partinfo> (E-P)

??

E. coli TOP10

<partinfo>pSB4C5</partinfo>

886 bp

Ligations 2010

BBa_J23100
_4C5

(it was <partinfo>BBa_J23100</partinfo>
_4C5-1)

<partinfo>BBa_J23100</partinfo>in <partinfo>pSB4C5</partinfo>.

27/08/2010

E. coli TOP10

<partinfo>pSB4C5</partinfo>

Length ok.

35 bp

Ligations 2010

BBa_J23101
_4C5

(it was <partinfo>BBa_J23101</partinfo>
_4C5-1)

<partinfo>BBa_J23101</partinfo> in <partinfo>pSB4C5</partinfo>.

27/08/2010

E. coli TOP10

<partinfo>pSB4C5</partinfo>

Length ok.

35 bp

Ligations 2010

BBa_J23105
_4C5

(it was <partinfo>BBa_J23105</partinfo>
_4C5-1)

<partinfo>BBa_J23105</partinfo> in <partinfo>pSB4C5</partinfo>.

27/08/2010

E. coli TOP10

<partinfo>pSB4C5</partinfo>

Length ok.

35 bp

Ligations 2010

BBa_J23106
_4C5

(it was <partinfo>BBa_J23106</partinfo>
_4C5-1)

<partinfo>BBa_J23106</partinfo> in <partinfo>pSB4C5</partinfo>.

27/08/2010

E. coli TOP10

<partinfo>pSB4C5</partinfo>

Length ok.

35 bp

Ligations 2010

BBa_J23110
_4C5

(it was <partinfo>BBa_J23110</partinfo>
_4C5-1)

<partinfo>BBa_J23110</partinfo> in <partinfo>pSB4C5</partinfo>.

27/08/2010

E. coli TOP10

<partinfo>pSB4C5</partinfo>

Length ok.

35 bp

Ligations 2010

BBa_J23114
_4C5

(it was <partinfo>BBa_J23114</partinfo>
_4C5-1)

<partinfo>BBa_J23114</partinfo> in <partinfo>pSB4C5</partinfo>.

27/08/2010

E. coli TOP10

<partinfo>pSB4C5</partinfo>

Length ok.

35 bp

Ligations 2010

BBa_J23116
_4C5

(it was <partinfo>BBa_J23116</partinfo>
_4C5-1)

<partinfo>BBa_J23116</partinfo> in <partinfo>pSB4C5</partinfo>.

30/08/2010

E. coli TOP10

<partinfo>pSB4C5</partinfo>

Length ok.

35 bp

Ligations 2010

BBa_J23118>
_4C5

(it was <partinfo>BBa_J23118</partinfo>
_4C5-1)

<partinfo>BBa_J23118</partinfo> in <partinfo>pSB4C5</partinfo>.

30/08/2010

E. coli TOP10

<partinfo>pSB4C5</partinfo>

Length ok.

35 bp

Ligations 2010
Strain name Description Arrival date Infos Resistance Quality control Notes Store

MG1655(Seq)
(MG)

E. coli wild type strain fully sequenced

28/06/2010

link

- Genome fully sequenced Strains 2010

MC1061
(MC)

E. coli strain

28/06/2010

link

- Strain used by JC Anderson et al. for integration Strains 2010

BW25141
(BW1)

E. coli pir+ strain

28/06/2010

link

- Actually replicates the conditional replication origin R6K LC pir Strains 2010

BW25142
(BW2)

E. coli pir116 strain

28/06/2010

link

- Actually replicates the conditional replication origin R6K HC pir Strains 2010

BW23473

E. coli pir+ strain

19/07/2010

link

- LC pir Strains 2010

BW23474

E. coli pir116 strain

28/06/2010

link

- Actually replicates the conditional replication origin R6K HC pir Strains 2010

BW5328/pAH123

E. coli strain

28/06/2010

link

Amp 50 ug/ml Plasmid length ok (after digestion with SpeI).

Contains pAH123 HELPER plasmid carrying IntPhi80, low copy Amp, ori-ts 30°C

Strains 2010

BT340

E. coli strain

28/06/2010

link

Amp 100 ug/ml and Cm 34 ug/ml Plasmid relative molecular weight ok (after gel run of the undigested plasmid)

Contains pCP20 helper with yeast FRT recombinase, Amp, Cm, ori-ts 30°C

Strains 2010

DSM15372

E. coli DMS15372 strain

08/07/2010

link

Amp 100 ug/ml Length ok (after digestion with EcoRI)

Harbours the pBHR68 plasmid (contains an IPTG-inducible phaCAB operon)

Strains 2010