Expression Construct Name: pCS2P+sp5l ORF

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Description: zebrafish sp5-like (sp5l) zinc-finger transcription factor of the SP5 family (cDNA clone 27i2 from the Wnt8 upregulated microarray screen) ORF only in pCS2P+. PCR amplified using primers 96 &97 from 27-i2 (pSPORT) clone and cloned EcoRI/XbaI into pCS2P+. Injection of high doses (>260 ng/µl corresponding to 200ng/µl #53.GFP) results in loss of eyes.



Species of Sequence Origin: zebrafish

Notes for Use: For functional synthetic RNA, linearize with NotI, SP6 (1385bp RNA). Select clones with amp.

Reference: Development, Current Biology papers from the lab (2005).

LOCUS        Gil156.pCS    5168 bp     DNA    circular              3-JUL-2003

SOURCE      

  ORGANISM  

COMMENT      This file is created by Vector NTI

            http://www.informaxinc.com/

COMMENT      VNTDATE|290544468|

COMMENT      VNTDBDATE|307208401|

COMMENT      VNTNAME|Gil156.pCS2P+27-i2 ORF|

COMMENT      VNTAUTHORNAME|Demo User|

FEATURES              Location/Qualifiers

     mRNA             1..1

                     /vntifkey="54"

                     /label=cap\site\of\CMV\prom

                     /note="base1 = cap site for sCMV IE94 promoter"

     CDS              complement(2664..3524)

                     /vntifkey="4"

                     /label=AmpR

                     /note="beta-lactamase"

     promoter         35..51

                     /vntifkey="30"

                     /label=SP6

                     /note="SP6 promoter"

     5'UTR            <64..78

                     /vntifkey="52"

                     /label=ßglobin\5'\RNA

                     /note="B-globin 5' mRNA sequence"

     promoter         complement(1196..1212)

                     /vntifkey="30"

                     /label=T7

                     /note="T7 promoter (reverse orientation)"

     polyA_site       1217..1412

                     /vntifkey="26"

                     /label=SV40\pA

                     /note="SV40 late polyadenylation site (Bam to Dra1)"

     promoter         complement(1466..1482)

                     /vntifkey="30"

                     /label=T3

                     /note="T3 promoter (reverse orientation)"

     rep_origin       3653..4113

                     /vntifkey="33"

                     /label=f1\ori

                     /note="f1 ori"

     enhancer         4182..>5168

                     /vntifkey="9"

                     /label=CMV\enh/prom

                     /note="simian CMV IE94 enhancer/promoter"

     misc_feature     105..107

                     /vntifkey="21"

                     /label=first\ATG

     misc_feature     146..158

                      /vntifkey="21"

                     /label=27i2\spMO\half\binding\site

     misc_feature     942..1016

                     /vntifkey="21"

                     /label=pfam00096,\zf-C2H2,\Zinc\finger,\C2H2\type

     CDS              105..1178

                     /vntifkey="4"

                     /label=SP-like\transcription\factor

BASE COUNT      1300 a       1337 c       1258 g       1273 t

ORIGIN

        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag

       61 ctacttgttc tttttgcagg atcccatcga ttcgaattcc caccatggct gcattaacct

      121 tatccagagc ggacaatttc ctccacaatt tcttacagga ccgcactccc agctcatctc

      181 cagaaagtgg tccaaacacc ctctcctttc tggccaccac atgcagtcag gcttggcagg

      241 aaggctcgca gctcccttac gagggtccgg tgggctccgc atccagcatg ttccagctct

      301 ggagcaacga cgtggccccg aactccagct taagcgctca ccagatgacc ttcaccgtgc

      361 ccaaaatgca gtttcccagc cacatgcaac ccactctggg ctcgcactcc catcaccacc

      421 atcaccacca tcatcaccac cacgagcttc ccctcactcc tccagcggag cctccgtctg

      481 catattcatt cgagctttcc ccggttaaaa tgcaagggaa tgcgccatac tacacgcagc

      541 acaacgccgt cagtcaaaac tttccaagct tcctccaaaa cccctcagga agagctcacc

      601 tgcctggagg tcacgttgag gatggccagc agtggtggag tctcccccag ggtaacagcg

       661 ctccctccgg gcaccctttc tccttgggac ggcagctggt tttgggccat cagcctcaaa

      721 ttgcagcact tctgcaggga acctccaaag gtctgttgag ctctactcgt cgctgtcgcc

      781 gatgcaaatg cccaaactgc cagtccacag gaaacggagg tgctgcattg gagtttggaa

      841 agaaaagact acacatttgt cacattccag attgtgggaa ggtttataaa aaaacctctc

      901 acctcaaagc gcacctgcgc tggcatgctg gtgaacggcc cttcatttgc aactggctct

      961 tctgcgggaa gagcttcacg cgttcagatg agctgcagag gcatctccgt acacacaccg

     1021 gggagaagcg tttcggctgt cagcagtgtg gaaaacggtt tatgcggagc gatcatctct

     1081 ccaagcatgt gaagacccat cagagccgga agagccggtc cagtcagcct cctcacagcg

     1141 ggactgatgc tctgctgagc aacatcaaga gagagtaatc tagattctgc agccctatag

     1201 tgagtcgtat tacgtagatc cagacatgat aagatacatt gatgagtttg gacaaaccac

     1261 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt

     1321 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt

     1381 tcaggttcag ggggaggtgt gggaggtttt ttaattcgcg gccgcggcgc caatgcattg

     1441 ggcccggtac ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca

     1501 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga

     1561 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt

     1621 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga

     1681 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc

     1741 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg

     1801 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc

     1861 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc

     1921 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga

     1981 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc

     2041 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat

     2101 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg

     2161 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc

     2221 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga

     2281 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact

     2341 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt

     2401 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag

     2461 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg

     2521 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa

     2581 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata

     2641 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg

     2701 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata

     2761 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg

     2821 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct

     2881 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt

     2941 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc

     3001 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga

     3061 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt

     3121 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc

     3181 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa

     3241 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca

     3301 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca

     3361 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct

     3421 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc

     3481 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa

     3541 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt

     3601 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg

     3661 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta

     3721 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt

     3781 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca

     3841 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa

     3901 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat

     3961 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag

     4021 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg

     4081 ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt

     4141 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gtcgaccata gccaattcaa

     4201 tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt gccaattcaa

     4261 tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca gccaattcaa

     4321 tatggcggac ttggcaccat gccaattcaa tatggcggac ttggcactgt gccaactggg

     4381 gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg tgccaagtcc

     4441 gccatattga attggcatgg tgccaataat ggcggccata ttggctatat gccaggatca

     4501 atatataggc aatatccaat atggccctat gccaatatgg ctattggcca ggttcaatac

     4561 tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt gacgtagata

     4621 gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac cgcccatagc

     4681 cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac gtcatatggg

     4741 cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta aatggcccgc

     4801 ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa tgggatggct

     4861 cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg taaatggccc

     4921 acttggcagt acatcaatat ctattaatag taacttggca agtacattac tattggaagg

     4981 acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg gcccgcgatg

     5041 gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa atgggcgttc

     5101 cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc aatgctcgtt

     5161 tagggaac

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