ID   CA542029   standard; RNA; EST; 389 BP.
XX
AC   CA542029;
XX
SV   CA542029.1
XX
DT   20-NOV-2002 (Rel. 73, Created)
DT   20-NOV-2002 (Rel. 73, Last updated, Version 1)
XX
DE   C0613G06-5N NIA Mouse Trophoblast Stem Cell cDNA Library (Long) Mus
DE   musculus cDNA clone NIA:C0613G06 IMAGE:30021485 5', mRNA sequence.
XX
KW   EST.
XX
OS   Mus musculus (house mouse)
OC   Eukaryota; Metazoa; Chordata; Craniata; Vertebrata; Euteleostomi; Mammalia;
OC   Eutheria; Rodentia; Sciurognathi; Muridae; Murinae; Mus.
XX
RN   [1]
RP   1-389
RA   Piao Y., Kargul G.J., Dudekula D.B., Qian Y., Lim M.K., Luo A., Tanaka T.,
RA   Kunath T., Rossant J., Ko M.S.H.;
RT   "Systematic Analyses of NIA Mouse Trophoblast Stem Cell cDNA Library
RT   (Long)";
RL   Unpublished.
XX
CC   Other_ESTs: C0613G06-3
CC   Contact: Dawood B. Dudekula
CC   Laboratory of Genetics
CC   National Institute on Aging/National Institutes of Health
CC   333 Cassell Drive, Suite 4000, Baltimore, MD 21224-6820, USA
CC   Email: cdna@lgsun.grc.nia.nih.gov
CC   Plate: C0613  row: G  column: 06
CC   Seq primer: M13 Reverse
CC   High quality sequence stop: 389
CC   POLYA=No.
XX
FH   Key             Location/Qualifiers
FH
FT   source          1..389
FT                   /db_xref="taxon:10090"
FT                   /note="Vector: pSPORT1 (Invitrogen); Site_1: SalI; Site_2:
FT                   NotI; Mouse cDNA project by the Laboratory of Genetics,
FT                   National Institute on Aging (NIA), Intramural Research
FT                   Program, NIH (http://lgsun.grc.nia.nih.gov/cDNA). This is a
FT                   long-transcript enriched cDNA library (Ref. Genome Res. 11:
FT                   1553-1558 (2001). [PMID: 11544199]). Total RNAs were
FT                   obtained from Dr. Janet Rossant and Tilo Kunath (Samuel
FT                   Lunenfeld Research Institute, Canada). Double-stranded
FT                   cDNAs were synthesized with an Oligo(dT) primer
FT                   [Invitrogen:
FT                   5'-pGACTAGTTCTAGATCGCGAGCGGCCGCCCTTTTTTTTTTTTTTT-3'] from 4
FT                   ug of total RNA, treated with T4 DNA polymerase, and
FT                   purified by ethanol-precipitation. The cDNAs were ligated
FT                   to Lone-linker LL-Sal4, purified by phenol/chloroform, and
FT                   separated from free linkers by Centricon 100. Then, the
FT                   cDNAs were amplified by long-range high fidelity PCR using
FT                   Ex Taq polymerase (Takara) with a primer Sal4-S. The
FT                   products were purified by phenol/chloroform and Centricon
FT                   100. The cDNAs were digested with SalI and NotI enzymes and
FT                   cloned into SalI/NotI site of pSPORT1 plasmid vector. The
FT                   DH10B E. coli host was transformed with the ligation
FT                   mixture by the standard chemical method. The average insert
FT                   size is about 2.6 kb. The library was constructed by Yulan
FT                   Piao (NIA)."
FT                   /organism="Mus musculus"
FT                   /strain="B5/EGFP transgenic ICR mice"
FT                   /clone="NIA:C0613G06 IMAGE:30021485"
FT                   /clone_lib="NIA Mouse Trophoblast Stem Cell cDNA Library
FT                   (Long)"
FT                   /tissue_type="Trophoblast stem cell"
FT                   /dev_stage="3.5-dpc"
FT                   /lab_host="DH10B"
XX
SQ   Sequence 389 BP; 108 A; 107 C; 94 G; 80 T; 0 other;
     cgtctcgtgc gtgttcattc agccacgatg cctgaggaaa cccagaccca agaccaacca        60
     atggaggagg aggaggtcga gacctttgcc tttcaggcag aaattgccca gttaatgtcc       120
     ttgatcatca ataccttcta ctcgaacaaa gagatctttc tgagggagct catctccaat       180
     tcatcggacg ctctggataa aatccgttac gagagcctga cggaccccag taaactggac       240
     tcggggaagg agctgcacat caatctcatt cccagcaaac aggaccgaac cctgaccatt       300
     gtggataccg ggattggaat gaccaaggcc gacttgatca ataaccttgg caccattgcc       360
     aagtcgggca ccaaagcctt catggaggc                                         389
//