f1月on f1 (-origin 21-327 B-galactosidase a-fragment 460-816 multiple cloning site 653-760 ampicillin lac promoter 817-938 clacZ -Sac I pUC origin 1158-1825 ampicillin resistance (bla)ORF 1976-2833 pBluescript Il KS (- MCS 3.0kb -Kpn I AP lac pBS KS- pUC ori pBluescript Il KS(+/-)Multiple Cloning Site Region (sequence shown 598-826) BssHlI T7 Promoter TTGTAAAACGACGGCCAGTGAGCGCGCGTAATACGACTCACTATAGGGCGAATTGGAGCTCCACCGCGGTGGCGGCCGCTCTAGA. M13-20 primer binding site T7 primer binding site SK primer binding site. Hinc l Spel Xho Dra ll .ACTAGTGGATCCCCCGGGCTGCAGGAATTCGATATCAAGCTTATCGATACCGTCGACCTCGAGGGGGGGCCCGGTACC. .primer bindingste KS primer binding site ←T3 Promoter -gal a-fragment .CAGCTTTTGTTCCCTTTAGTGAGGGTTAATTGCGCGCTTGGCGTAATCATGGTCATAGCTGTTTCC T3 primer binding site M13Reverse primer bindingsite12 pBS KS-