Team:Evry/Freezer

From 2012.igem.org

(Difference between revisions)
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<tr style="background: #ccccff; text-align: center;">
<tr style="background: #ccccff; text-align: center;">
-
<td width="5%"> Number
+
<td width="5%"> Code
</td><td width="10%"> Name
</td><td width="10%"> Name
</td><td width="25%"> Sequence
</td><td width="25%"> Sequence
Line 411: Line 411:
<tr style="background: #ccccff; text-align: center;">
<tr style="background: #ccccff; text-align: center;">
-
<td width="10%"> Name
+
<td width="5%"> Code
-
</td><td width="100"> Sequence
+
</td><td width="5%"> Name
</td><td width="5%"> Length
</td><td width="5%"> Length
 +
</td><td width="5%"> Concentration (ng/ul)
</td><td width="5%"> Species of origin
</td><td width="5%"> Species of origin
</td><td width="5%"> Expression domain
</td><td width="5%"> Expression domain
</td><td width="5%"> Restriction sites (EcoRI, XbaI, SpeI, PtsI)
</td><td width="5%"> Restriction sites (EcoRI, XbaI, SpeI, PtsI)
</td><td width="5%"> Reference
</td><td width="5%"> Reference
 +
</td><td width="100"> Sequence
<tr style="background: #cccccc;">
<tr style="background: #cccccc;">
-
<td style="background: #ccffcc;">p2 Xlurp
+
<td style="background: #ccffcc;">Promo 1
-
</td><td width="100">tctgtgtctccaaagatgccccagtagctccccatcttattttcactgcacatgctttgtactgctgtcacttactgagctcagggacccactcacaatatacagtacacatagaaatttcacagtataaggctgattagtaattaatacagataattactacatggaattacagaaaccagtgcaattagcatcagaatttaataatcagccctgtagcatcagcttatatgacaggccaacctcattttctgcttgataatttgcaacaactcctacgcttagcttctcaacagctgctcagagcccactgagcatgtgagagtcacagacactttccaagatggtgaccccctgtgacaggtttgaagtcctggatcattgctgctattgacaagctgaaactttaggctggtgcaataagttcagtatataaaacattgcaattctagccataataatttttagggtttagttctcctttaaggtggggccttgctaaagatgacttatttgcacatttataacatctgtcttaaaatagcatcttataactcatatacaaaaaaggggttatattgctacctacatgtcactcagcattaaaaagttccatgttccatgtttaagaggacatatttacaccgtttcaaacagtgaaatattctcatcttgctcgttggcaagaatgaggttaagtttgcagtgttgaataaaagataaatagtgattagctaaatttttcggcaggtttcattgtgaaaaagatgcccataggtgaaaaaaactgtgcgcaacaaaaaaattaattgtcgcccttcgccttcaatgcatttactgaccttttggcgaaacgagtcacattcgcccatcactaaagataaattaagtagtacacttccttcagtcttaaagtgcatcaacccaatgcaagatcatattgagctagaaggggaaggaatgccatatcatggataacacagggagataaatgtatctcctgtttgtagtagaggtcctatccatcagtaaaatatgtagacctctgcttcctagtggtaattgttgtacgcctcaacatccgtttctcaactagcaaagctttctcaaagcttattaccacctggagattaagcacctagatctagttattttccttgtgattgtgatatacagtatatgggacatcctagttggtaatttgcactacaggggaacaaaatggcatattcaggaatatatcagaaagaaaagcacatgcaaatagtgacaaacaatgcactgttaaaaaacaaaaaaagagaggcaggcctgttactctaggaaattcacctttaagagggagggagatataatttactaatatcagttaatttatttgaaagagaaaaagtagtaaaccccccactctctttaatacagtataaaaaaatattaaacagattgtgaacacaattaatgtggtgttaattcatatttgtgaattggtgtacaaacattcattgattaatcaaagtgcaagttcttcaatagtgcaatcaatgtccatgattttaagattaattaattatcttaaaaaaattattcaattagttcaatttttaaaagctgtaccctgcttgtaatctaattaactaaattcatctaataagtggaattaccgagtgtaattacagaaggtagcaattatattgaattaacaatatcaaattgaattcattaatttaaagtgctagtgcaaatataattaattagaaggtgaattaattcaatgaaagttaaaaatgtatgtccaggaccaacatctgaaaaaatgtaggaaaaggaaaataatagaggtggagaggtcgcaaaaacttctataagaggtttttctatgccctgggactccacacaatggagaagacaaacactggggactgtggtctcaaattaaccgtatctatctcattgtggagaggctaagctgtactataaaaccacagatccttcagtgtcattgaaagtaaggagaatttaaaggaagagaaaatcaaatgcggcacttgaaaggctaacagcgctggatattaattaaaagaaccaagccaattctaaaaagggtttggtagagaacagatttgttaaaatcaatttgaaagttaggagactccgccaatgcactgtaataagaaaaagtatatcagtttgcaggaaaacggagaaggcaaaactgggtggctaaaagaagtgagaaaaagaaggactagaaatgaaggggtggaaacctgagtggtgtagtaaaaagcaacaataaataaaaaaacacaacatagtgaaaatattttattgccatctgtttattaaaatctgtctattctgcacctgcaattctaaaatcattgtattctgaacagctgatattgcattgctctgttggaacttgaatggcggccctgtggctatttaagcttaattctataaacaatattagaaactcgaaaatgaacacacagaatttaccagtagaggattaaagtttattttatgtaaatgaatacaaaattcttactgtttttggttgctattggccgtttagatattttttaagcaaaacattagcaaagccctcaaagttaataattcatcggttaggccgtcctttattgatccatcaatcaggattcagttttgtgctgtcaaaaacaatgtgcgtgggaacaactttatgtggagcagggactcttcatgctctggtcagaaaatgtacaggaacttggaacagagaagaagtataaagaaaataatagcatgcatcacaatgtttctggtgtgccaatgaatagctgtgtgttacaaaatctctgtcttgtataaagacaacaccagctacagtaagttaaattctaaacacatatttgtccaatacaggggcccaattaatactgcttgtgcttttacggcacaatcaatctttaagaaaacccctcaagagtttcctaactcacgctggttttcatgctgaaaaagaaagctttgtaagttacccgacctgctgcaggactttatcccagggtgtattaataggagtttttacaataatggctcatggtaatactctgattaaactcaatagctcattgcaatactttgataaagaactgtataaaaatataacacaaagaactgttctgctgttgaacccgatgtcgtatgttgagcccaatatagagtaaaaagacagctttataacgtataattgtgtataacaaagctttgtatgatagtatacaaggcagccttataatccataatcctgtataatgaagcgttgtgcaagagaaatcatgtattatctaatatgcttatgatttctgaagcccttatcagactcttctttattataaataggggcacaatatgctattcagctaccagctagctaagataagatctctgtttcggatgtgcttgtgggaaacaagatctgttattaaccaaatcattatgagttattgactaagcctctcaatgagcttactgtttgaaagcatatataaactgggggtcaccatttggtcagttgagatctcacttgtggggttgacgcgtgccgccacaggaccttccgtgctccgtttgagggattcaatctgctgactgttccaggatctcccagacttggttgcagattgtttgctttgctcttttactcattcggtaatgtatatcttcttcattctatgtatgtaataaacctttgcttatactagctggtgtgtttgaattacttccgagagactcatagaaccacaattaaaacacagggtaacaaagtctgggcacccctgagctgacatagggcatctattgtctttgtccacagtcctgatatttttttatcaagcagtaactatgggcatgcaatttgcagtggggaaaacaagaatttggattggtgctagaactgtgtgctgctgaacaacaatgaaaactgccatcttaattctgctgaactgacacagctattttcttttttaaatatgtctattagttaaggggttaaactttaaaatgatgtagagagtaatatcccgagacaatttgctattggtttttattttttttactatttatggtatttgagctatttcgctttttattcagcagctctccagtttgcaattctggcgatctggttattaggatccaatgggtcagtgacccctgtctgaaagctgaaaagcctcctaagaagtaggcaagtcatttaaaaaacgatataaaaaaataaaataatgaagaccaagtgaaaagttgcttagaagtggccaatctatatcatacttaaagggatattgtcatgggaaaaacatttttttcaaaatgaatcagttaatagtgctgctccatcagaattctgcactgaaatccatttctcaaaagagcaaacagatttttttatattcaattttgaaatttgacatggggctagacatattgtcaatttcccagctgccccaagtcatgtgacttgtgctctgataaactttaatcactctttactgctgtactgcaagttagagtgatatcaccccctccctttttccccacagcagccaaacaaaagaacaaagggaaggtaatcagatagcagctccttaacacaagataacagctgcctggttagatataagaacaacactcaatagtaaaaacccatgtcccactgagacacattcagttacattgagaaggaaaaacagcagcctgccagaaagcatttctctctaaagtgcaggcacaagtcacatgacttggggcagctgggaaattgacaaaatgtctagccccatgtcagatttcaaaattgaatataaaaaaatctgtttgctcttttgagaaatggatttcagtgcaaaattctgctggagaagcactattaactatttgaaaaaaacatgttttctgatgacagtatccctttaaagttgacttcatggtaaaccacccctttgaaaacattacagactgtcagaaaagtatgtataaatggggaaaatatcaggaccacctaatttgagatttggtaggtatgtcactgtttgttacttttttcggttagtgaccttgcaacatttcctgtaacagcttatttcagtgtttacccttgttgatcaagggttacacaaattcccagaaatataacgtgcctttagtgattgtgcaatccatatagataaaaggggatcttgctggcagagttcacagaagagctgacttagaaacccact
+
</td><td>p2 Xlurp
</td><td>5273bp
</td><td>5273bp
 +
</td><td>177,1
</td><td>Xenopus Laevis
</td><td>Xenopus Laevis
</td><td>Myeloid cells
</td><td>Myeloid cells
</td><td>/
</td><td>/
</td><td>Smith et al., 2002
</td><td>Smith et al., 2002
 +
</td><td>Sequence
 +
<tr style="background: #cccccc;">
 +
<td style="background: #ccffcc;">Promo 2
 +
</td><td>p2 CMV
 +
</td><td>1016bp
 +
</td><td>65,3
 +
</td><td>Cytomegalovirus
 +
</td><td>Widespread
 +
</td><td>/
 +
</td><td>Werdien et al., 2001
 +
</td><td>Sequence
 +
 +
 +
<tr style="background: #cccccc;">
 +
<td style="background: #ccffcc;">Promo 3
 +
</td><td>p2 Flk-1
 +
</td><td>2723bp
 +
</td><td>54
 +
</td><td>Xenopus Laevis
 +
</td><td>Vasculature
 +
</td><td>/
 +
</td><td>Doherty et al., 2007
 +
</td><td>Sequence
 +
 +
 +
<tr style="background: #cccccc;">
 +
<td style="background: #ccffcc;">Promo 4
 +
</td><td>p2 14xUAS E1b-ATG
 +
</td><td>454bp
 +
</td><td>93,7
 +
</td><td>
 +
</td><td>
 +
</td><td>Xba1
 +
</td><td>
 +
</td><td>Sequence
 +
 +
 +
<tr style="background: #cccccc;">
 +
<td style="background: #ccffcc;">Promo 5
 +
</td><td>p2 ROSA26
 +
</td><td>814bp
 +
</td><td>55,7
 +
</td><td>Mus musculus
 +
</td><td>Widespread
 +
</td><td>/
 +
</td><td>Gross et al., 2006
 +
</td><td>Sequence
 +
 +
 +
<tr style="background: #cccccc;">
 +
<td style="background: #ccffcc;">Promo 6
 +
</td><td>p2 foxi short
 +
</td><td>2020bp
 +
</td><td>105,9
 +
</td><td>Xenopus Tropicalis
 +
</td><td>Ionocytes
 +
</td><td>/
 +
</td><td>Unpublished results
 +
</td><td>Sequence
 +
 +
 +
<tr style="background: #cccccc;">
 +
<td style="background: #ccffcc;">Promo 7
 +
</td><td>p2 UASt
 +
</td><td>341bp
 +
</td><td>64,8
 +
</td><td>Saccharomyces cerevisiae
 +
</td><td>GAL4 inductible
 +
</td><td>/
 +
</td><td>
 +
</td><td>Sequence
 +
 +
<tr style="background: #cccccc;">
 +
<td style="background: #ccffcc;">Promo 8
 +
</td><td>p2 HB9
 +
</td><td>3142bp
 +
</td><td>48,1
 +
</td><td>Danio rerio
 +
</td><td>Motor neurons
 +
</td><td>PtsI
 +
</td><td>Flanagan-Street et al., 2005
 +
</td><td>Sequence
 +
 +
<tr style="background: #cccccc;">
 +
<td style="background: #ccffcc;">Promo 9
 +
</td><td>p2 pax6
 +
</td><td>bp
 +
</td><td>58,9
 +
</td><td>
 +
</td><td>
 +
</td><td>
 +
</td><td>
 +
</td><td>Sequence
 +
 +
<tr style="background: #cccccc;">
 +
<td style="background: #ccffcc;">Promo 10
 +
</td><td>p2 NBT
 +
</td><td>3481bp
 +
</td><td>168,7
 +
</td><td>Xenopus Laevis
 +
</td><td>Differentiated neurons
 +
</td><td>EcoRI
 +
</td><td>Huang et et al., 2007
 +
</td><td>Sequence
 +
 +
<tr style="background: #cccccc;">
 +
<td style="background: #ccffcc;">Promo 11
 +
</td><td>p2 LMO2 long
 +
</td><td>4730bp
 +
</td><td>170,1
 +
</td><td>
 +
</td><td>
 +
</td><td>
 +
</td><td>
 +
</td><td>Sequence
 +
 +
<tr style="background: #cccccc;">
 +
<td style="background: #ccffcc;">Promo 12
 +
</td><td>p2 pax3
 +
</td><td>2916bp
 +
</td><td>109,5
 +
</td><td>Xenopus Laevis
 +
</td><td>Neural ectoderm
 +
</td><td>
 +
</td><td>Unpublished results
 +
</td><td>Sequence
 +
 +
<tr style="background: #cccccc;">
 +
<td style="background: #ccffcc;">Promo 13
 +
</td><td>p2 vimentin
 +
</td><td>7205bp
 +
</td><td>187,4
 +
</td><td>Xenopus Tropicalis
 +
</td><td>Neural progenitor/glia
 +
</td><td>
 +
</td><td>Love et al., 2011
 +
</td><td>Sequence
 +
 +
<tr style="background: #cccccc;">
 +
<td style="background: #ccffcc;">Promo 14
 +
</td><td>p2 Brachyury
 +
</td><td>3856bp
 +
</td><td>103,6
 +
</td><td>
 +
</td><td>
 +
</td><td>
 +
</td><td>
 +
</td><td>Sequence
 +
 +
<tr style="background: #cccccc;">
 +
<td style="background: #ccffcc;">Promo 15
 +
</td><td>p2 CMG long
 +
</td><td>
 +
</td><td>92,4
 +
</td><td>
 +
</td><td>
 +
</td><td>
 +
</td><td>
 +
</td><td>Sequence
 +
 +
<tr style="background: #cccccc;">
 +
<td style="background: #ccffcc;">Promo 16
 +
</td><td>p2 LMO2 short
 +
</td><td>2007bp
 +
</td><td>88,3
 +
</td><td>
 +
</td><td>
 +
</td><td>
 +
</td><td>
 +
</td><td>Sequence
 +
 +
<tr style="background: #cccccc;">
 +
<td style="background: #ccffcc;">Promo 17
 +
</td><td>p2 Ef1a
 +
</td><td>528bp
 +
</td><td>128,9
 +
</td><td>
 +
</td><td>
 +
</td><td>
 +
</td><td>
 +
</td><td>Sequence
 +
 +
<tr style="background: #cccccc;">
 +
<td style="background: #ccffcc;">Promo 18
 +
</td><td>p2 Hsp70
 +
</td><td>487bp
 +
</td><td>62,5
 +
</td><td>Xenopus Laevis
 +
</td><td>Heat shock inductible
 +
</td><td>PrsI
 +
</td><td>Beck et al., 2003
 +
</td><td>Sequence
 +
 +
<tr style="background: #cccccc;">
 +
<td style="background: #ccffcc;">Promo 19
 +
</td><td>pCAR (eGFP)
 +
</td><td>bp
 +
</td><td>2900
 +
</td><td>
 +
</td><td>
 +
</td><td>
 +
</td><td>
 +
</td><td>Sequence
</table>
</table>

Revision as of 14:35, 6 August 2012


Freezer


Primers


Code Name Sequence Length Tm  %GC Comments Date
P1 Tir1-BB-FW cgcttctagATGACGTACTTCCCGGAGG 19 57 58 13th july
P2 Tir1-BB-Rv ctactagtaTTATTATAGGATTTTAACAAAATTTGGTGC 24 52 29 13th july
P3 SdMTir1-BB-Fw AACTGAACTCACTACTGTACTTCTGTCACCAAATGACTAATGCTGCTCTAGTTACTGTCGCCAAG 65 72 43 13th july
P4 SdMTir1-BB-Rv TTGGACAGCCCAAGGATACTGCAACAAGTCCCTCCTCTGTCACAGCAGAATAACCAGCTACG 62 76 52 13th july
P5 Tir1 SdM v02-Fw tgcTgtgacagaggagggac 20 60 60 25th july
P6 Tir1 SdM v02-Fw ctagAgcagcattagtcatttgg 23 55 43 25th july
P7 pCS2+BB-Fw cgctgcagGAACTATAGTGAGTCGTATTACG 23 51 39 13th july
P8 pCS2+BB-Rv tgcatGAATTCGAATCGATGGGATCC 21 54 48 13th july
P9 GFP-AID-BB-Fw gaattcgcggccgcttctagagATGGCCACAACCATGGTG 40 74 58 20th july
P10 GFP-AID-BB-Rv tactagtagcggccgctgcagTTATTAAACCTTACGTTTCTTTTTAGGGAC 51 70 43 20th july
P11 p2 Hsp70-Fw ACTGTCGACCCCGTTTAGCAGGAAATAGCC 30 67 53 Heat Shock 20th july
P12 p2 Hsp70-Rv ACTAAGCTTATTTGCGCTCCTTACAGTTTGC 31 63 42 Heat Shock 20th july
P13 p2 Flk-1-Fw ACTGTCGACCTGGACTCTGGGCTAGGG 27 68 63 Vasculature 20th july
P14 p2 Flk-1-Rv TCAAAGCTTGTTGACGTTTAGTCCAGGACAG 31 64 45 Vasculature 20th july
P15 p2 ROSA-Fw ACTGTCGACTAGATGAAGGAGAGCC 25 61 52 Ubiquitous 20th july
P16 p2 ROSA-Rv GATAAGCTTGGATCCCCGCAAACGCAC 27 66 56 Ubiquitous 20th july
P17 p2 CarA-Fw ACTGTCGACGTCCCATACCAGTACAATGCAAT 32 66 47 muscles 20th july
P18 p2 CarA-Rv AGTAAGCTTGTGCTGTGATTGAATTGGCTG 30 63 43 muscles 20th july
P19 IaaH-BB-Fw gaattcgcggccgcttctagATGCGCGAAATGATTACGC 19 55 47 23th july
P20 IaaH-BB-Rv ctgcagcggccgctactagtaTTATTAGCCTTTTAACACTTTTTCG 25 52 28 23th july
P21 IaaM-BB-Fw gaattcgcggccgcttctagatgtttggaccggatttccc 20 56 50 23th july
P22 IaaM-BB-Rv ctgcagcggccgctactagtattattagtcccccagcgc 39 55/73 56/59 23th july
P23 AID-Fw cgcttctagGGAGCTGGTGCAGGCGCT 18 64 72 25th july
P24 Sp6 ATTTAGGTGACACTATA 17 41 27 for BB detection when sequenced NP
P25 T3 AATTAACCCTCACTAAAGGG 20 50 40 for BB detection when sequenced NP
P26 primer_sequencing_pCS2+_Fw CAAGTGTAGCGGTCACGCTG 20 59 60 13th july
P27 primer_sequencing_pCS2+_Rv GCGTAATCATGGTCATAGCTG 21 52 48 13th july
P28 GFP_AID_BB_Fw_v2 TGCATgaattcgcggccgcttctagagATGGCCACAACCATGGTG 45 75 56
P29 IaaH_BB_Fw_v2 TGCATgaattcgcggccgcttctagATGCGCGAAATGATTACGC 44 74 52
P30 IaaM_BB_Fw_v2 TGCATgaattcgcggccgcttctagatgtttggaccggatttccc 45 74 53
P31 IRES_BB_F GTTTCTTCGAATTCGCGGCCGCTTCTAGAGCGATCATTCAagagaTCCCC 50 53/74 50/52
P32 IRES_BB_R GTTTCTTCCTGCAGCGGCCGCTACTAGTAttgtggccatattatcatcg 49 51/72 40/49
P33 BBa_G1005 gtttcttcctgcagcggccgctactagta 29 67 55
P34 IaaM_Kozak_BB_F GTTTCTTCGAATTCGCGGCCGCTTCTAGAGGGTTCCAAACCATGCGCGAAATGATTACGC 60 55/76 46/52
P35 IaaH_Kozak_BB_F GTTTCTTCGAATTCGCGGCCGCTTCTAGAGGGTTCCAAACCatgtttggaccggatttcc 60 54/76 47/52


Promoteurs


Code Name Length Concentration (ng/ul) Species of origin Expression domain Restriction sites (EcoRI, XbaI, SpeI, PtsI) Reference Sequence
Promo 1 p2 Xlurp 5273bp 177,1 Xenopus Laevis Myeloid cells / Smith et al., 2002 Sequence


Promo 2 p2 CMV 1016bp 65,3 Cytomegalovirus Widespread / Werdien et al., 2001 Sequence


Promo 3 p2 Flk-1 2723bp 54 Xenopus Laevis Vasculature / Doherty et al., 2007 Sequence


Promo 4 p2 14xUAS E1b-ATG 454bp 93,7 Xba1 Sequence


Promo 5 p2 ROSA26 814bp 55,7 Mus musculus Widespread / Gross et al., 2006 Sequence


Promo 6 p2 foxi short 2020bp 105,9 Xenopus Tropicalis Ionocytes / Unpublished results Sequence


Promo 7 p2 UASt 341bp 64,8 Saccharomyces cerevisiae GAL4 inductible / Sequence
Promo 8 p2 HB9 3142bp 48,1 Danio rerio Motor neurons PtsI Flanagan-Street et al., 2005 Sequence
Promo 9 p2 pax6 bp 58,9 Sequence
Promo 10 p2 NBT 3481bp 168,7 Xenopus Laevis Differentiated neurons EcoRI Huang et et al., 2007 Sequence
Promo 11 p2 LMO2 long 4730bp 170,1 Sequence
Promo 12 p2 pax3 2916bp 109,5 Xenopus Laevis Neural ectoderm Unpublished results Sequence
Promo 13 p2 vimentin 7205bp 187,4 Xenopus Tropicalis Neural progenitor/glia Love et al., 2011 Sequence
Promo 14 p2 Brachyury 3856bp 103,6 Sequence
Promo 15 p2 CMG long 92,4 Sequence
Promo 16 p2 LMO2 short 2007bp 88,3 Sequence
Promo 17 p2 Ef1a 528bp 128,9 Sequence
Promo 18 p2 Hsp70 487bp 62,5 Xenopus Laevis Heat shock inductible PrsI Beck et al., 2003 Sequence
Promo 19 pCAR (eGFP) bp 2900 Sequence









Glycerol Stocks


Number Strain Plasmid Description Date
P1 E.Coli DH5 aplpha pCS2+ RFP ( BBA_J23100 ) 13th july
P2 E.Coli DH5 aplpha pCS2+ Auxine () 13th july
P3 E.Coli DH5 aplpha pCS2+ Tir1 GFP-Aid () 19th july
P4 E.Coli DH5 aplpha pCS2+ Tir1 GFP-Aid () 19th july


Retrieved from "http://2012.igem.org/Team:Evry/Freezer"