Zebrafish EST vector


Clones are streaked onto LB agar containing ampicillin (50µg/ml). To use: re-streak the clone onto the same agar type, in such a way as to obtain single colonies. Incubate at 37°C overnight. Clones can be grown in LB broth + ampicillin, and glycerol added after growth to a concentration of 25% for long-term storage at -70°C.

cDNA was oligo dT primed from 8 ug of total RNA isolated from FACS sorted GFP positive myoblast cells taken from 3-9 somite stage Tg(alpha-actin:GFP) embryos. SalI_NotI cut cDNA was then ligated into pSPORT1 with SalI at the 5' end and NotI at the 3' end.

zebrafish plasmid

 

1 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta 60

61 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg 120

121 ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgaatttagg tgacactata 180

181 gaagagctat gacgtcgcat gcacgcgtac gtaagcttgg atcctctaga gcggccgccg 240

241 actagtgagc tcgtcgaccc gggaattccg gaccggtacc tgcaggcgta ccagctttcc 300

301 ctatagtgag tcgtattaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa 360

361 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 420

421 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 480

481 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 540

541 tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat 600

601 tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca 660

661 gcaggcgaaa atcctgtttg atggtggttg acggcgggat ataacatgag ctgtcttcgg 720

721 tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac tcggtaatgg 780

781 cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc 840

841 cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc 900

901 gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca 960

961 gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg 1020

1021 cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga 1080

1081 tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca 1140

1141 cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg acccgttgcg 1200

1201 cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca 1260

1261 ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg 1320

1321 gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca 1380

1381 gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt 1440

1441 cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag 1500

1501 agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc accaccctga 1560

1561 attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg 1620

1621 tgtcaacgta aatgccgctt cgccttcgcg cgcgaattgc aagctctgca ttaatgaatc 1680

1681 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact 1740

1741 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 1800

1801 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 1860

1861 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 1920

1921 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 1980

1981 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 2040

2041 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc 2100

2101 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 2160

2161 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 2220

2221 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 2280

2281 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 2340

2341 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 2400

2401 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 2460

2461 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 2520

2521 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 2580

2581 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 2640

2641 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 2700

2701 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 2760

2761 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 2820

2821 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 2880

2881 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 2940

2941 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 3000

3001 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 3060

3061 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 3120

3121 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 3180

3181 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 3240

3241 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 3300

3301 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 3360

3361 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 3420

3421 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 3480

3481 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 3540

3541 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 3600

3601 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgaaattgta 3660

3661 aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac 3720

3721 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg 3780

3781 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa 3840

3841 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt 3900

3901 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt 3960

3961 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga 4020

4021 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc 4080

4081 gcgcttaatg cgccgctaca gggcgcgtc 4109