2966

WO2010099296 Hyperactive PiggyBac transposases

catatgaagctTAATACGACTCACTATAagGATTAAT

pfizer 5'UTR

gagaataaactagtattcttctggtccccacagactcagagagaaccc

Kozak

gccacc

ATGGGCTCTAGCCTGGACGACGAGCACATCCTGAGCGCCCTGCTGCAGAGCGACGACGAACTGGTGGGCGAGGACAGCGACAGCGAGATCAGCGACCACGTGTCCGAGGACGACGTGCAGTCCGACACCGAGGAAGCCTTCATCGACGAGGTGCACGAAGTGCAGCCTACCAGCAGCGGCTCCGAGATCCTGGACGAGCAGAACGTGATCGAGCAGCCTGGCAGCTCCCTGGCCAGCAACAGAATCCTGACCCTGCCCCAGAGAACCATCAGAGGCAAGAACAAGCACTGCTGGTCCACCTCCAAGAGCACCAGGCGGAGCAGAGTGTCCGCCCTGAACATCGTGCGGAGCCAGAGGGGCCCCACCAGAATGTGCAGAAACATCTACGACCCCCTGCTGTGCTTCAAGCTGTTCTTCACCGACGAGATCATCAGCGAGATCGTGAAGTGGACCAACGCCGAGATCAGCCTGAAGAGGCGGGAGAGCATGACCGGCGCCACCTTCAGAGACACCAACGAGGACGAGATCTACGCCTTCTTCGGCATCCTGGTGATGACCGCCGTGAGAAAGGACAACCACATGAGCACCGACGACCTGTTCGACAGATCCCTGAGCATGGTGTACGTGTCCGTGATGAGCAGAGACAGATTCGACTTCCTGATCAGATGCCTGAGAATGGACGACAAGAGCATCAGACCCACCCTGCGGGAGAACGACGTGTTCACCCCCGTGCGGAAGATCTGGGACCTGTTCATCCACCAGTGCATCCAGAACTACACCCCTGGCGCCCACCTGACCATCGATGAGCAGCTGCTGGGCTTCAGAGGCAGATGCCCCTTCAGAATGTACATCCCCAACAAGCCCAGCAAGTACGGCATCAAGATCCTGATGATGTGCGACAGCGGCACCAAGTACATGATCAAC

change GGCATGC to GGaATGC to remove Sph1

GGaATGC

CCTACCTGGGCAGAGGCACCCAGACAAACGGCGTGCCCCTGGGCGAGTACTACGTGAAAGAACTGAGCAAGCCTGTGCATGGCAGCTGCAGGAACATCACCTGCGACAACTGGTTCACCAGCATCCCCCTGGCCAAGAACCTGCTGCAGGAACCCTACAAGCTGACCATCGTGGGCACCGTGCGGAGCAACAAGCGGGAGATCCCAGAGGTGCTGAAGAACAGCAGATCCAGACCTGTGGGAACAAGCATGTTCTGCTTCGACGGCCCCCTGACCCTGGTGTCCTACAAGCCCAAGCCCGCCAAGATGGTGTACCTGCTGTCCAGCTGCGACGAGGACGCCAGCATCAACGAGAGCACCGGCAAGCCCCAGATGGTGATGTACTACAACCAGACCAAGGGCGGCGTGGACACCCTGGACCAGATGTGCAGCGTGATGACCTGCAGCAGAAAGACCAACAGATGGCCCATGGCCCTGCTGTACGGCATGATCAATATCGCCTGCATCAACAGCTTCATCATCTACAGCCACAACGTGTCCAGCAAGGGCGAGAAGGTGCAGAGCCGGAAGAAATTCATGCGGAACCTGTACATGAGCCTGACCTCCAGCTTCATGAGAAAGAGACTGGAAGCCCCCACCCTGAAGAGATACCTGCGGGACAACATCAGCAACATCCTGCCCAACGAAGTGCCAGGAACAAGCGACGACAGCACCGAGGAACCCGTGATGAAGAAGAGGACCTACTGCACCTACTGTCCCAGCAAGATCAGAAGAAAGGCCAACGCCAGCTGCAAGAAATGCAAAAAAGTGATCTGCCGGGAGCACAACATCGACATGTGCCAGAGCTGTTTC

Stop

TGA

TGA
3’ UTR 
Xho1 Not1
CTCGAG CGGCCGC
change C to g to remove Sph1 site
CTGGTACTGCATGgACGCAATGCTAGCT GCCCCTTTCCCGTCCTGGGTACCCCGAGTCTCCCCCGACCTCGGGTCCCAGGTATGCTCCCACCTCCACCTGCCCCACTCACCACCTCTGC TAGTTCCAGACACCTCC

CAAGCACGCAGCAATGCAGCTCAAAACGCTTAGCCTAGCCACACCCCCACGGGAAACAGCAGTGATTAACCTTT AGCAATAAACGAAAGTTTAACTAAGCTATACTAACCCCAGGGTTGGTCAATTTCGTGCCAGCCAC

ACCCTGGAGCTAGCA 

Sph1

GCATGC

==

sequence for gene synthesis

CATATGAAGCTTAATACGACTCACTATAAGGATTAATGAGAATAAACTAGTATTCTTCTGGTCCCCACAGACTCAGAGAGAACCCGCCACCATGGGCTCTAGCCTGGACGACGAGCACATCCTGAGCGCCCTGCTGCAGAGCGACGACGAACTGGTGGGCGAGGACAGCGACAGCGAGATCAGCGACCACGTGTCCGAGGACGACGTGCAGTCCGACACCGAGGAAGCCTTCATCGACGAGGTGCACGAAGTGCAGCCTACCAGCAGCGGCTCCGAGATCCTGGACGAGCAGAACGTGATCGAGCAGCCTGGCAGCTCCCTGGCCAGCAACAGAATCCTGACCCTGCCCCAGAGAACCATCAGAGGCAAGAACAAGCACTGCTGGTCCACCTCCAAGAGCACCAGGCGGAGCAGAGTGTCCGCCCTGAACATCGTGCGGAGCCAGAGGGGCCCCACCAGAATGTGCAGAAACATCTACGACCCCCTGCTGTGCTTCAAGCTGTTCTTCACCGACGAGATCATCAGCGAGATCGTGAAGTGGACCAACGCCGAGATCAGCCTGAAGAGGCGGGAGAGCATGACCGGCGCCACCTTCAGAGACACCAACGAGGACGAGATCTACGCCTTCTTCGGCATCCTGGTGATGACCGCCGTGAGAAAGGACAACCACATGAGCACCGACGACCTGTTCGACAGATCCCTGAGCATGGTGTACGTGTCCGTGATGAGCAGAGACAGATTCGACTTCCTGATCAGATGCCTGAGAATGGACGACAAGAGCATCAGACCCACCCTGCGGGAGAACGACGTGTTCACCCCCGTGCGGAAGATCTGGGACCTGTTCATCCACCAGTGCATCCAGAACTACACCCCTGGCGCCCACCTGACCATCGATGAGCAGCTGCTGGGCTTCAGAGGCAGATGCCCCTTCAGAATGTACATCCCCAACAAGCCCAGCAAGTACGGCATCAAGATCCTGATGATGTGCGACAGCGGCACCAAGTACATGATCAACGGAATGCCCTACCTGGGCAGAGGCACCCAGACAAACGGCGTGCCCCTGGGCGAGTACTACGTGAAAGAACTGAGCAAGCCTGTGCATGGCAGCTGCAGGAACATCACCTGCGACAACTGGTTCACCAGCATCCCCCTGGCCAAGAACCTGCTGCAGGAACCCTACAAGCTGACCATCGTGGGCACCGTGCGGAGCAACAAGCGGGAGATCCCAGAGGTGCTGAAGAACAGCAGATCCAGACCTGTGGGAACAAGCATGTTCTGCTTCGACGGCCCCCTGACCCTGGTGTCCTACAAGCCCAAGCCCGCCAAGATGGTGTACCTGCTGTCCAGCTGCGACGAGGACGCCAGCATCAACGAGAGCACCGGCAAGCCCCAGATGGTGATGTACTACAACCAGACCAAGGGCGGCGTGGACACCCTGGACCAGATGTGCAGCGTGATGACCTGCAGCAGAAAGACCAACAGATGGCCCATGGCCCTGCTGTACGGCATGATCAATATCGCCTGCATCAACAGCTTCATCATCTACAGCCACAACGTGTCCAGCAAGGGCGAGAAGGTGCAGAGCCGGAAGAAATTCATGCGGAACCTGTACATGAGCCTGACCTCCAGCTTCATGAGAAAGAGACTGGAAGCCCCCACCCTGAAGAGATACCTGCGGGACAACATCAGCAACATCCTGCCCAACGAAGTGCCAGGAACAAGCGACGACAGCACCGAGGAACCCGTGATGAAGAAGAGGACCTACTGCACCTACTGTCCCAGCAAGATCAGAAGAAAGGCCAACGCCAGCTGCAAGAAATGCAAAAAAGTGATCTGCCGGGAGCACAACATCGACATGTGCCAGAGCTGTTTCTGATGACTCGAGCGGCCGCCTGGTACTGCATGGACGCAATGCTAGCTGCCCCTTTCCCGTCCTGGGTACCCCGAGTCTCCCCCGACCTCGGGTCCCAGGTATGCTCCCACCTCCACCTGCCCCACTCACCACCTCTGCTAGTTCCAGACACCTCCCAAGCACGCAGCAATGCAGCTCAAAACGCTTAGCCTAGCCACACCCCCACGGGAAACAGCAGTGATTAACCTTTAGCAATAAACGAAAGTTTAACTAAGCTATACTAACCCCAGGGTTGGTCAATTTCGTGCCAGCCACACCCTGGAGCTAGCAGCATGC

MGSSLDDEHILSALLQSDDELVGEDSDSEISDHVSEDDVQSDTEEAFIDEVHEVQPTSSGSEILDEQNVIEQPGSSLASNRILTLPQRTIRGKNKHCWSTSKSTRRSRVSALNIVRSQRGPTRMCRNIYDPLLCFKLFFTDEIISEIVKWTNAEISLKRRESMTGATFRDTNEDEIYAFFGILVMTAVRKDNHMSTDDLFDRSLSMVYVSVMSRDRFDFLIRCLRMDDKSIRPTLRENDVFTPVRKIWDLFIHQCIQNYTPGAHLTIDEQLLGFRGRCPFRMYIPNKPSKYGIKILMMCDSGTKYMINGMPYLGRGTQTNGVPLGEYYVKELSKPVHGSCRNITCDNWFTSIPLAKNLLQEPYKLTIVGTVRSNKREIPEVLKNSRSRPVGTSMFCFDGPLTLVSYKPKPAKMVYLLSSCDEDASINESTGKPQMVMYYNQTKGGVDTLDQMCSVMTCSRKTNRWPMALLYGMINIACINSFIIYSHNVSSKGEKVQSRKKFMRNLYMSLTSSFMRKRLEAPTLKRYLRDNISNILPNEVPGTSDDSTEEPVMKKRTYCTYCPSKIRRKANASCKKCKKVICREHNIDMCQSCF

genewiz optimized

ATGGGCTCTTCACTGGATGACGAGCACATCTTGTCGGCACTGCTGCAGAGCGATGACGAA CTGGTGGGAGAGGACAGCGACAGTGAGATATCTGACCACGTGTCTGAGGATGATGTGCAG TCTGACACCGAGGAGGCGTTCATTGACGAAGTCCATGAGGTGCAACCGACCTCCTCCGGT TCGGAGATCCTGGACGAACAGAACGTGATTGAGCAACCAGGCTCCTCTCTCGCTAGCAAT CGAATCCTCACCCTGCCGCAGCGGACCATCCGCGGAAAAAACAAGCACTGTTGGTCAACG TCCAAGTCTACGCGGCGCTCGAGGGTCTCGGCACTAAACATAGTACGATCCCAGCGCGGG CCCACTAGGATGTGCCGCAACATCTACGATCCGTTGCTGTGCTTCAAACTATTTTTCACC GACGAGATCATCTCTGAGATCGTGAAATGGACGAACGCCGAGATTTCTCTGAAGCGCCGC GAGAGTATGACCGGCGCCACGTTCAGGGATACCAACGAGGACGAGATCTACGCCTTCTTC GGAATCCTGGTCATGACAGCCGTGCGCAAGGACAACCATATGTCGACGGATGACCTTTTT GACCGCTCTCTGTCCATGGTGTACGTGTCTGTGATGTCGCGGGACCGCTTCGACTTCCTG ATCCGCTGCCTGCGCATGGATGACAAGTCGATCCGCCCCACACTCAGAGAGAACGACGTG TTCACCCCGGTGCGAAAGATCTGGGACTTGTTCATCCACCAGTGTATCCAGAACTACACC CCTGGCGCGCACCTGACCATTGATGAGCAGCTGCTCGGCTTCCGTGGCCGTTGTCCCTTC CGCATGTATATTCCCAACAAGCCATCGAAGTACGGCATTAAGATCCTGATGATGTGCGAC AGCGGTACCAAGTACATGATCAACGGTATGCCCTATCTGGGGCGGGGCACCCAGACCAAC GGCGTTCCGCTTGGGGAGTACTACGTGAAGGAGCTGTCCAAGCCCGTGCATGGTTCCTGC CGAAACATCACCTGTGACAACTGGTTTACCTCTATCCCTCTGGCGAAGAACCTGCTCCAG GAGCCATACAAACTGACCATCGTTGGTACCGTGAGGTCCAATAAGCGCGAGATCCCGGAG GTTCTGAAGAATTCCCGCTCCCGCCCCGTGGGGACTTCCATGTTCTGCTTTGACGGCCCT TTGACCCTTGTGAGCTACAAGCCCAAACCTGCCAAGATGGTCTACTTACTGAGTAGTTGC GACGAAGATGCTAGCATCAATGAGAGCACTGGCAAGCCTCAGATGGTGATGTATTACAAC CAGACTAAAGGCGGCGTCGACACTCTAGACCAGATGTGCTCTGTAATGACGTGCTCCCGC AAAACAAATCGTTGGCCCATGGCTCTCCTGTATGGAATGATCAATATCGCCTGTATCAAC TCCTTCATCATCTACTCCCACAACGTGTCCTCTAAGGGCGAGAAGGTCCAGTCCCGGAAG AAATTTATGCGTAATCTTTACATGTCACTTACCTCCAGCTTTATGAGAAAGCGCCTGGAG GCTCCTACCCTTAAGCGTTACTTGCGCGACAACATCAGCAACATCCTGCCTAACGAGGTA CCCGGGACTAGCGATGACTCTACTGAGGAGCCCGTCATGAAAAAACGTACTTACTGTACC TACTGCCCATCCAAGATTCGTCGTAAGGCCAACGCGTCCTGCAAGAAGTGCAAGAAGGTC ATCTGCCGCGAGCACAACATTGACATGTGCCAGAGCTGCTTC

MGSSLDDEHILSALLQSDDELVGEDSDSEISDHVSEDDVQSDTEEAFIDEVHEVQPTSSGSEILDEQNVIEQPGSSLASNRILTLPQRTIRGKNKHCWSTSKSTRRSRVSALNIVRSQRGPTRMCRNIYDPLLCFKLFFTDEIISEIVKWTNAEISLKRRESMTGATFRDTNEDEIYAFFGILVMTAVRKDNHMSTDDLFDRSLSMVYVSVMSRDRFDFLIRCLRMDDKSIRPTLRENDVFTPVRKIWDLFIHQCIQNYTPGAHLTIDEQLLGFRGRCPFRMYIPNKPSKYGIKILMMCDSGTKYMINGMPYLGRGTQTNGVPLGEYYVKELSKPVHGSCRNITCDNWFTSIPLAKNLLQEPYKLTIVGTVRSNKREIPEVLKNSRSRPVGTSMFCFDGPLTLVSYKPKPAKMVYLLSSCDEDASINESTGKPQMVMYYNQTKGGVDTLDQMCSVMTCSRKTNRWPMALLYGMINIACINSFIIYSHNVSSKGEKVQSRKKFMRNLYMSLTSSFMRKRLEAPTLKRYLRDNISNILPNEVPGTSDDSTEEPVMKKRTYCTYCPSKIRRKANASCKKCKKVICREHNIDMCQSCF