Team:UNIPV-Pavia/Freezer

From 2009.igem.org

EthanolPVanimation.gif

Biobrick/
Ligation Name
Description Creation Date Strain Vector Quality Control
J23100(J100)
replaced by "J23100 Jun4"
constitutive promoter May 25 TOP10 J61002(Amp) sequence ok (MIT)
J23101(J101)
replaced by "J23101 Jun4"
constitutive promoter May 25 TOP10 J61002(Amp) sequence ok (MIT)
J23102(J102) constitutive promoter May 25 TOP10 J61002(Amp) sequence ok (MIT)
J23103(J103) constitutive promoter May 25 TOP10 J61002(Amp) sequence ok (MIT)
J23104(J104) constitutive promoter May 25 TOP10 J61002(Amp) sequence ok (MIT)
J23105(J105) constitutive promoter May 25 TOP10 J61002(Amp) sequence ok (MIT)
J23106(J106) constitutive promoter May 25 TOP10 J61002(Amp) sequence ok (MIT)
J23110(J110) constitutive promoter May 25 TOP10 J61002(Amp) sequence ok (MIT)
J23112(J112) constitutive promoter May 25 TOP10 J61002(Amp) sequence ok (MIT)
J23113(J113) constitutive promoter May 25 TOP10 J61002(Amp) sequence ok (MIT)
J23114(J114) constitutive promoter May 25 TOP10 J61002(Amp) sequence ok (MIT)
J23116(J116) constitutive promoter May 25 TOP10 J61002(Amp) sequence ok (MIT)
J23117(J117) constitutive promoter May 25 TOP10 J61002(Amp) sequence ok (MIT)
J23118(J118)
replaced by "J23118 Jun4"
constitutive promoter May 25 TOP10 J61002(Amp) sequence ok (MIT)
E0240(GFP3) GFP protein generator
(RBS-GFP-TT)
May 25 TOP10 pSB1A2 length ok (PCR) sequence ok (MIT)
K131009(E2) colE2 operon:ceaB-ceiB-Tweak-celB-T (with RBS) May 28 TOP10 pSB1AK3 sequence BAD in prefix,suffix,celB (MIT,BMR)
K117000(celB) colE7 lysis gene May 28 TOP10 pSB1A2 sequence ok (MIT)
K124017(lysis21) Phage Lysis Cassette May 28 TOP10 pSB1AK3 sequence BAD (MIT)
B0015(TT) Double terminator May 28 TOP10 pSB1AK3 sequence ok (MIT)
I732017 (RBS-lacZ) RBS-lacZ May 28 TOP10 pSB1A2 long part (MIT)
B0030 RBS 0.6 Jun 4 TOP10 pSB1A2 sequence ok (MIT,BMR)
B0031 RBS 0.07 Jun 4 TOP10 pSB1A2 sequence ok (MIT)
B0032 RBS 0.3 Jun 4 TOP10 pSB1A2 sequence ok (MIT)
B0033 RBS 0.01 Jun 4 TOP10 pSB1A2 sequence ok (MIT)
B0034 RBS 1 Jun 4 TOP10 pSB1A2 sequence ok (MIT)
P0440(P04) RBS-tetR-TT Jun 4 TOP10 pSB1A2 length ok (PCR) sequence ok (MIT)
R0040(Ptet) Ptet Jun 4 TOP10 pSB1A2 sequence ok (MIT)
T9002 Ptet-RBS-luxR-TT-Plux-RBS-GFP-TT Jun 4 TOP10 pSB1A3 sequence BAD (MIT) - sequence ok (BMR)
J23100(J100 Jun4) constitutive promoter Jun 4 TOP10 J61002(Amp) sequence ok (MIT,BMR)
J23101(J101 Jun4) constitutive promoter Jun 4 TOP10 J61002(Amp) sequence ok (MIT)
J23118(J118 Jun4) constitutive promoter Jun 4 TOP10 J61002(Amp) sequence ok (MIT)
A1 (BBa_K173000) J23100(Jun 4)-E0240
(Pcon-RBS-GFP-TT)
Jun 11 TOP10 pSB1A2 length ok (PCR)
A2 (BBa_K173001) J23101(Jun 4)-E0240
(Pcon-RBS-GFP-TT)
Jun 11 TOP10 pSB1A2 length ok (PCR)
A3-1 (BBa_K173004) I732017-B0015
(RBS-lacZ-TT)
Jun 12 TOP10 pSB1AK3 length ok (digestion PstI) sequence ok -long part- (BMR)
A4 (BBa_K173005) P0440-R0040
(RBS-tetR-TT-Ptet)
Jun 11 TOP10 pSB1A2 length ok (PCR) sequence ok (BMR)
A5 (BBa_K173006) A4-A3
(RBS-tetR-TT-Ptet-RBS-lacZ-TT)
Jun 19 TOP10 pSB1AK3 sequence ok -long part- (BMR)
A6 (BBa_K173007) J23100(Jun 4)-A4
(Pcon-RBS-tetR-TT-Ptet)
Jun 19 TOP10 pSB1A2 length ok (PCR) sequence ok (BMR)
BOL1 J23118-B0034-C0012-B0015
(Pcon-RBS-lacI-TT)
Jun 23 TOP10 pSB1A2 sequence ok (BMR)
A7 (BBa_K173002) J23118(Jun 4)-E0240
(Pcon-RBS-GFP-TT)
Jun 25 TOP10 pSB1A2 length ok (PCR)
R0011(Plac) Plac Jun 25 TOP10 pSB1A2 sequence ok (MIT)
K112808(ENTERO) holin-lysozyme-T-Pconweak-antiholin-T (with RBSs) Jun 25 TOP10 pSB1A2 sequence almost ok -long part- (MIT): it has several gaps in the scars, a silent point mutation in holin gene and a wrong annotated terminator at the end of the brick (that is actually K112710, not B0010).
A8 A4-E0240
(RBS-tetR-TT-Ptet-RBS-GFP-TT)
Jun 29 TOP10 pSB1A2 NON PURE PLASMID! (digestion E-P) but works
A9 A6-E0240
(Pcon-RBS-tetR-TT-Ptet-RBS-GFP-TT)
Jun 29 TOP10 pSB1A2 NON PURE PLASMID! (digestion E-P) but works
A10
thrown away
K117000-B0015
(celB-TT)
Jun 30 TOP10 pSB1AK3 sequence BAD (BMR)
A8pg (BBa_K173008) A4-E0240
(RBS-tetR-TT-Ptet-RBS-GFP-TT)
Jul 3 TOP10 pSB1A2 length ok (digestion E-P)
A9pg (BBa_K173009) A6-E0240
(Pcon-RBS-tetR-TT-Ptet-RBS-GFP-TT)
Jul 3 TOP10 pSB1A2 length ok (digestion E-P)
A11-1
thrown away and replaced
BOL1-R0011
(Pcon-RBS-lacI-TT-Plac)
Jul 10 TOP10 pSB1A2 length ok (PCR)
sequence MUTATED
A11-3
thrown away and replaced
BOL1-R0011
(Pcon-RBS-lacI-TT-Plac)
Jul 10 TOP10 pSB1A2 length ok (PCR)
A11-6
thrown away and replaced
BOL1-R0011
(Pcon-RBS-lacI-TT-Plac)
Jul 10 TOP10 pSB1A2 length ok (PCR)
F2620
thrown away
Ptet-RBS-luxR-TT-Plux
Jul 10 TOP10 pSB1A2 sequence BAD (MIT, BMR)
A12-2 (BBa_K173011) J23118(Jun 4)-A4
Jul 16 TOP10 pSB1A2 length ok (PCR)
sequence ok (BMR)
A12-3
it is a backup of A12-2
J23118(Jun 4)-A4
Jul 16 TOP10 pSB1A2 length ok (PCR)
sequence ok (BMR)
A14-4
thrown away
A11-E0240
Jul 16 TOP10 pSB1A2 NON PURE PLASMID! (digestion) and the A11 sample used for ligation was MUTATED (BMR)
A14pg
thrown away
A11-E0240
Jul 16 TOP10 pSB1A2 NON PURE PLASMID! (digestion) and the A11 sample used for ligation was MUTATED (BMR)
K116001 (PnhaA) PnhaA-RBS-GFP-TT
Jul 29 NEB 10-beta pSB1A2 sequence WRONG! it is actually K116002!!!
K116002 (PnhaA+GFP) PnhaA-RBS-GFP-TT
Jul 29 NEB 10-beta pSB1A2 sequence WRONG! it is actually J33204!!!
K112405 (Pcad) cadAB promoter
Jul 29 NEB 10-beta K112951(Amp+Cm) sequence ok (BMR)
P0412 RBS-lacI-TT
Jul 29 DH5alpha pSB1A2 sequence ok (BMR)
I746902 GFPmut3+his tag generator under Pbad
Jul 29 NEB 10-beta pSB1AK3 sequence ok -long part- (BMR)
I746903 GFPmut3+his tag generator under PT7
Jul 29 NEB 10-beta pSB1AK3 sequence almost ok (BMR): the RBS is B0030 and not B0034 as documented, but the function is the same
K101017
to throw away!
MioC promoter
Jul 29 NEB 10-beta pSB1A2 sequence BAD (BMR)
F2620MIT1 Ptet-RBS-luxR-TT-Plux
Jul 29 D1210(lacIq) pSB1A2 sequence ok (BMR)
F2620MIT2 Ptet-RBS-luxR-TT-Plux
Jul 29 D1210(lacIq) pSB1A2 sequence ok (BMR)
MRGENE1 (BBa_K173016) pdc Jul 17 TOP10 pMK-RQ
(Kan)
antibiotic ok
length ok (digestion X-P)
sequence ok (Mr Gene)
MRGENE1-2 pdc Jul 17 TOP10 pMK-RQ
(Kan)
antibiotic ok
sequence ok (Mr Gene)
MRGENE1-3 pdc Jul 17 TOP10 pMK-RQ
(Kan)
antibiotic ok
sequence ok (Mr Gene)
MRGENE2 (BBa_K173017) adhB Jul 17 TOP10 pMA
(Amp)
antibiotic ok
length ok (digestion X-P)
sequence ok (Mr Gene)
MRGENE2-2 adhB Jul 17 TOP10 pMA
(Amp)
antibiotic ok
sequence ok (Mr Gene)
MRGENE2-3 adhB Jul 17 TOP10 pMA
(Amp)
antibiotic ok
sequence ok (Mr Gene)
B1-13 (BBa_K173013) B0030-MRGENE1
(RBS-pdc)
Jul 28 TOP10 pSB1A2 length ok (digestion E-P
, digestion S-P and digestion E-S)
sequence ok (BMR)
B2-5 (BBa_K173014) B0030-MRGENE2
(RBS-adhB)
Jul 23 TOP10 pSB1A2 length ok (PCR
and digestion S-P)
sequence ok (BMR)
B3-5 (BBa_K173018) B1-B0015
(RBS-pdc-TT)
Jul 31 TOP10 pSB1AK3 length ok+extra band (digestion XhoI-P
and digestion X-P)
sequence ok (BMR)
B4-2 (BBa_K173019) B2-B0015
(RBS-adhB-TT)
Jul 31 TOP10 pSB1AK3 length ok (digestion XhoI-P)
sequence ok (BMR)
B5-3 B1-B4
(RBS-pdc-RBS-adhB-TT)
Aug 12 TOP10 pSB1AK3 length ok+2 extra bands (digestion E-P)
sequence ok (BMR)
B6-3 (BBa_K173020) B2-B3
(RBS-adhB-RBS-pdc-TT)
Aug 12 TOP10 pSB1AK3 length ok+2 extra bands (digestion E-P)
sequence ok (BMR)
A11-2
thrown away
BOL1-R0011
(Pcon-RBS-lacI-TT-Plac)
Aug 13 TOP10 pSB1A2 length ok (digestion S-P) sequence MUTATED (BMR)
A11-3 (BBa_K173010) BOL1-R0011
(Pcon-RBS-lacI-TT-Plac)
Aug 13 TOP10 pSB1A2 length ok (digestion S-P) sequence ok (BMR)
A15-1
it is a backup copy of A15-3
F2620MIT1-K112808
(Ptet-RBS-luxR-TT-Plux-RBS-holin-RBS-lysozyme-T-Pconweak-RBS-antiholin-T)
Aug 14 TOP10 pSB1A2 length ok (digestion) sequence ok (BMR)
A15-2
thrown away
F2620MIT1-K112808
(Ptet-RBS-luxR-TT-Plux-RBS-holin-RBS-lysozyme-T-Pconweak-RBS-antiholin-T)
Aug 14 TOP10 pSB1A2 length ok (digestion) sequence WRONG (BMR)
A15-3 (BBa_K173015) F2620MIT1-K112808
(Ptet-RBS-luxR-TT-Plux-RBS-holin-RBS-lysozyme-T-Pconweak-RBS-antiholin-T)
Aug 14 TOP10 pSB1A2 length ok (digestion) sequence ok (BMR)
B8-5
thrown away
A4-B6
(RBS-tetR-TT-Ptet-RBS-adhB-RBS-pdc-TT)
Aug 21 TOP10 pSB1AK3 length ??? (digestion E-P) sequence WRONG (BMR)
A14-3
to throw away!
A11(-2 Aug13)-E0240
(Pcon-RBS-lacI-TT-Plac-RBS-GFP-TT)
Aug 21 TOP10 pSB1A2 length ok (PCR) sequence of A11-2(Aug13) was MUTATED (BMR)
A16-4 (BBa_K173026) A12-E0240
(Pcon-RBS-tetR-TT-Ptet-RBS-GFP-TT)
Aug 21 TOP10 pSB1A2 length ok (PCR) sequence ok (BMR)
A17-4
to throw away!
R0011-E0240
(Plac-RBS-GFP-TT)
Aug 28 TOP10 pSB1A2 length ok (PCR) sequence WRONG! ligation failed.
A18
thrown away
A15(-2)-pSB3K3
(Ptet-RBS-luxR-TT-Plux-RBS-holin-RBS-lysozyme-T-Pconweak-RBS-antiholin-T)
Aug 28 TOP10 pSB3K3 not screened, its parent plasmid (A15-2) was wrong
K116002 (PnhaA+GFP) PnhaA-RBS-GFP-TT
Sep 4 TOP10 pSB1A2 sequence almost ok (BMR): inconsistent prefix (but restriction sites are good) and additional "c" at the end of promoter
B5new2-3 (BBa_K173003) B1-B4 (RBS-pdc-RBS-adhB-TT) Sep 1 TOP10 pSB1AK3 length ok (digestion E-P), sequence ok -long part- (BMR)
B9-2 (BBa_K173021) F2620-B5new2(-3) (Ptet-RBS-luxR-TT-Plux-RBS-pdc-RBS-adhB-TT) Sep 4 TOP10 pSB1AK3 length ok (digestion E-P), sequence ok -long part- (BMR)
B10-2 (BBa_K173022) F2620-B6 (Ptet-RBS-luxR-TT-Plux-RBS-adhB-RBS-pdc-TT) Sep 4 TOP10 pSB1AK3 length ok + 2 extra bands (digestion E-P), sequence ok -long part- (BMR)
B9new-1 to 5 F2620-B5new2(-3) (Ptet-RBS-luxR-TT-Plux-RBS-pdc-RBS-adhB-TT) Oct 1 TOP10 pSB1AK3 length ok (digestion E-P)
A19 F2620 Oct 1 TOP10 pSB4C5 length ok (digestion E-P)
A21 A19-B5new2(-3) Oct 8 TOP10 pSB4C5 length ok (digestion E-P)
A14S-3 (BBa_K173012) J23118-RBS-lacI-TT-Plac-RBS-GFP-TT (A11(-3)-E0240) ? TOP10 pSB1A2 length ok (digestion E-P)
A17new (BBa_K173025) Plac-RBS-GFP-TT (R0011-E0240) Sep 4 TOP10 pSB1A2 length ok (PCR), sequence ok (BMR)
A20 F2620-A8 (Ptet-RBS-luxR-TT-Plux-RBS-tetR-TT-Ptet-RBS-GFP-TT) Oct 8 TOP10 pSB1A2 length ok (digestion E-P)
I52002 (4C5) plasmid ? TOP10 pSB4C5 length ok (digestion E-P)
I714891 (3K3) plasmid Aug 21 TOP10 pSB3K3 length ok (digestion E-P)
B14 (BBa_K173023) F2620-B3 (Ptet-RBS-luxR-TT-Plux-RBS-pdc-TT) Oct 16 TOP10 pSB1A2 to screen
B15 (BBa_K173024) F2620-B4 (Ptet-RBS-luxR-TT-Plux-RBS-adhB-TT) Oct 16 TOP10 pSB1A2 to screen