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a putative dominantly activating version of the zebrafish Spr2 (sp5-like, sp5l, 27i2) transcription factor. VP16 activator domain fused to the N-terminus of the C-terminal 136aa of zebrafish Spr2 (27i2), which contain the zinc finger domain and are better conserved to other SP factors than the N-terminus, ergo likely comprise the DNA-binding domain. suitable for transcribing sense RNA stabilized by the Xenopus globin UTRs. VP16 amplified from 227 using primers 278 & 279; Spr2 amplified from 156 using primers 280 & 281. Triple ligation VP16 EcoRI/AscI, Spr2 AscI/SacII into 54 EcoRI/SacII. Sequenced and found good.
Notes for Use: For functional synthetic RNA cut XbaI, T3 (1000bp RNA) Select clones with amp.
Reference: Development, Current Biology papers from the lab (2005).
LOCUS Gil233.RN3 3831 bp DNA circular 12-DEC-2003
SOURCE
ORGANISM
COMMENT This file is created by Vector NTI
http://www.informaxinc.com/
COMMENT VNTDATE|321822360|
COMMENT VNTDBDATE|321823105|
COMMENT VNTNAME|Gil233.RN3 VP16-Spr2|
COMMENT VNTAUTHORNAME|Gilbert Weidinger|
COMMENT VNTAUTHOREML|gweidin@u.washington.edu|
FEATURES Location/Qualifiers
promoter complement(1020..1039)
/vntifkey="30"
/label=T7
/note="T7 promoter"
promoter 1..20
/vntifkey="30"
/label=T3
/note="T3 promoter"
3'UTR 43..87
/vntifkey="50"
/label=Xenopus\globin\5'\UTR
CDS 1790..2650
/vntifkey="4"
/label=AmpR
misc_signal 940..982
/vntifkey="87"
/label=poly\A-C
3'UTR 792..982
/vntifkey="50"
/label=Globin\3'\UTR
CDS 105..341
/vntifkey="4"
/label=VP16\activator\domain
CDS 342..362
/vntifkey="4"
/label=linker
misc_structure 372..713
/vntifkey="88"
/label=putative\DNA-binding\domain\(conserved)
CDS 372..779
/vntifkey="4"
/label=C-terminal\136aa\of\Spr2
BASE COUNT 1010 a 963 c 970 g 888 t
ORIGIN
1 aattaaccct cactaaaggg aacaaaagct ggagctaagc ttgcttgttc tttttgcaga
61 agctcagaat aaacgctcaa ctttggcaga tctgaattcc caccatggcc cccccgaccg
121 atgtcagcct gggggacgag ctccacttag acggcgagga cgtggcgatg gcgcatgccg
181 acgcgctaga cgatttcgat ctggacatgt tgggggacgg ggattccccg gggccgggat
241 ttacccccca cgactccgcc ccctacggcg ctctggatat ggccgacttc gagtttgagc
301 agatgtttac cgatgccctt ggaattgacg agtacggtgg gggaggaagc ggacccggag
361 gaaggcgcgc ccgctgtcgc cgatgcaaat gcccaaactg ccagtccaca ggaaacggag
421 gtgctgcatt ggagtttgga aagaaaagac tacacatttg tcacattcca gattgtggga
481 aggtttataa aaaaacctct cacctcaaag cgcacctgcg ctggcatgct ggtgaacggc
541 ccttcatttg caactggctc ttctgcggga agagcttcac gcgttcagat gagctgcaga
601 ggcatctccg tacacacacc ggggagaagc gtttcggctg tcagcagtgt ggaaaacggt
661 ttatgcggag cgatcatctc tccaagcatg tgaagaccca tcagagccgg aagagccggt
721 ccagtcagcc tcctcacagc gggactgatg ctctgctgag caacatcaag agagagtaat
781 taattaaccg cggatctggt taccactaaa ccagcctcaa gaacacccga atggagtctc
841 taagctacat aataccaact tacactttac aaaatgttgt cccccaaaat gtagccattc
901 gtatctgctc ctaataaaaa gaaagtttct tcacattcta aaaaaaaaaa aaaaaaaaaa
961 aaaaaaaacc cccccccccc cctgcagggc caagtcggcc tctagaggta cccaattcgc
1021 cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
1081 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
1141 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
1201 ggaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
1261 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
1321 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
1381 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
1441 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
1501 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
1561 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
1621 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
1681 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
1741 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
1801 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
1861 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
1921 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
1981 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
2041 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
2101 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
2161 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
2221 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
2281 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
2341 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
2401 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
2461 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
2521 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
2581 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
2641 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
2701 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
2761 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
2821 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
2881 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
2941 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
3001 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
3061 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
3121 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
3181 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
3241 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
3301 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
3361 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
3421 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
3481 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
3541 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
3601 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
3661 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
3721 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
3781 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctcg a
//