pIN4 5671 31 EcoO109 14 XbaI 371 NruI 408 EcoRI 456 HindIII 463 BamHI 469 Bpu1102 476 DraIII 611 AgeI 943 TthI 1050 Asp718 1068 KpnI 1068 EcoNI 1198 BstEII 1962 Bsp120I 1988 BssHII 2192 EcoRV 2229 Esp3I 2390 NarI 2420 SexAI 2640 AccI 2749 Eco47-3 3473 SapI 3694 DrdI 3919 Eam1105 4705 BsaI 4777 FspI 4930 PvuI 5077 ScaI 5188 XmnI 5305 AatII 5628 6 lpp-lac PO>promoter sequence 1 304 cw N ompAompA signal peptide 392 454 cw N lpp trmlipoprotein gene trx terminator 477 758 cw N lac Ilac I gene 1353 2576 cw N pUC ORIorigin of replication 3998 3823 ccw N Ampbeta-lactamase 5500 4637 ccw N 0 sequence TAGGCGTATCACGAGGCCCTTTGGATAACCAGAAGCAATAAAAAATCAAATCGGATTTCACTATATAATCTCACTTTATCTAAGATGAATCCGATGGAAGCATCCTGTTTTCTCTCAATTTTTTTATCTAAAACCCAGCGTTCGATGCTTCTTTGAGCGAACGATCAAAAATAAGTGCCTTCCCATCAAAAAAATATTCTCAACATAAAAAACTTTGTGTAATACTTGTAACGCTACATGGAGATTAACTCAATCTAGCTAGAGAGGCTTTACACTTTATGCTTCCGGCTCGTATAATGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGGATTCACTGGAACTCTAGATAACGAGGGCAAAAAATGAAAAAGACAGCTATCGCGATTGCAGTGGCACTGGCTGGTTTCGCTACCGTAGCGCAGGCCGGAATTCCAAGCTTGGATCCGGCTGAGCAACGACGTGAACGCAATGCGTTCCGACGTTCAGGCTGCTAAAGATGACGCAGCTCGTGCTAACCAGCGTCTGGACAACATGGCTACTAAATACCGCAAGTAATAGTACCTGTGAAGTGAAAAATGGCGCACATTGTGCGACATTTTTTTTGTCTGCCGTTTACCGCTACTGCGTCACGCGTAACATATTCCCTTGCTCTGGTTCACCATTCTGCGCTGACTCTACTGAAGGCGCATTGCTGGCTGCGGGAGTTGCTCCACTGCTCACCGAAACCGGATACCCTGCCCGACGATACAACGCTTTATCGACTAACTTCTGATCTACAGCCTTATTGTCTTTAAATTGCGTAAAGCCTGCTGGCAGTGTGTATGGCATTGTCTGAACGTTCTGCTGTTCTCCTGCCGATAGTGGTCGATGTACTTCAACATAACGCATCCCGTTAGGCTCCACGGAATATTTCACCGGTTCGTTGATCACTTTCACCGGCGTTCCCGTCCGCACGCTGGAGAACAAGGCTTTAATATCCGGTGCATTCATGCGAATACACCCTGAACTGACGCGCAAACCGACGCTGTCCGGCGCACTGGTACCATGAATGAGGTATTCGCCATTACCATGCGCGAGGCGCAGTGCGTAACGTCCTAGCGGGTTATTTGGTCCGGCAGGAACGACTGGCGGTAATTTAATGCCACGCTCCAGCGAACGCTGACGAATGCCTGCCGTAGGCGTCCAGGTTGGGTTAGGGATTTTCTGCCCAACACGCGTTTCCATCACCGGCGTTTCCAGCCCCTGCAATCCAATACCTATTGGATAAACCTGCACAATATTTTCTCCCGGCGGATAATAATAAAGGCGCAGCTCTGCAAGGTTGACACCATCGAATGGCGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTCAATTCAGGGTGGTGAATGTGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTCAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTAACGATAATCCCCTGACGCGGTGCATCAGGTAATAACAGTTGTGAAGGAATAGTTATCGTCGTACCAGGTTTTGGCACCGGGGCGATAGTGTTATTGGCTTCAAGGATCAACATTGCCGCAGTATCAAAACGTCGGGCAATAGCCTGAAGGTTTTTATCCCCTTCTTGCACCGTATACGTTTGATTTTGCCCAACCAGTCGGCTTCCGGTTGGTGGTAGCGGATAATCAACCGCCCAGGCAGCCTGGATGGCGCTAAAAGCGCCGATAAGCGTGAGTGTAAGCAAAGACGCGCGTTTCATTGTAAACCTCCTGTATTTGCCGGAGACTCACGCTGAAACGTCGGATGGCGCTTATGTTCACCTGAAACCAAAACACTCCTGTGCAGGTCAGTGTAAACATTGACCATCCGGCAATGTGAGCCAACCGGATGAAAGCTGTCCTTTTAGTTTAGCTAAGTGCAGCGGCTTTGGCGCGAATTGCGCGAATCATCGCTTCCAGACCTTGTGAACGAGATGGGGTGAGATGTTGGGTGAGCGCCATTTTTTCAAACCACGGACGCACATCGAAATTGACAATATCCTGCGGCGTCATCTGATCGTAGAGAATAAAGACGACCGCAATAAGCCCTTTCACAATCGCCGCATCGCTGTCGCCCTGTAATTCAATAATTCCCTGGGCATTCTGGCGCATGACAATCCACACCTGACTCTGACAGCCCTGAATGCTATTTTGTGGACTTCTGTCTTCGTCGCGTAATTCTGGCAGACGCTGGGGGACCGATGCCCTTGAGAGCCTTCAACCCAGTCAGCTCCTTCCGGTGGGCGCGGGGCATGACTATCGTCGCCGCACTTATGACTGTCTTCTTTATCATGCAACTCGTAGGACAGGTGCCGGCAGCGCTCTGGGTCATTTTCGGCGAGGACCGCTTTCGCTGGAGCGCGACGATGATCGGCCTGTCGCTTGCGGTATTCGGAATCTTGCACGCCCTCGCTCAAGCCTTCGTCACTGGTCCCGCCACCAAACGTTTCGGCGAGAAGCAGGCCATTATCGCCGGCATGGCGGCCCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTTCCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTAGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGGACAGTATTTGGTATCTGCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTTCGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCCCCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGCCGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGGGAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGGTGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATCCCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCAGTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTTCTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCCGGCGTCAACACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCTTCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCAACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGCAAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGCATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGGTTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTCTAAGAAACCATTATTATCATGACATTAACCTATAAAAA description High copy pIN expression vector annotations ; Constructed by Dr. Shinyu Chang by digesting pIN3 with Eag 1 ;+ (Xma 3), filling in with Klenow, and recutting with Pvu 1. ;+ Vector isolated and ligated with Pvu 1-Pvu 2 fragment of pJ ;+L2E1, which contains the pUC origin of replication.