pLEXSY_I-neo3 EGE-245 Description: nt 0001-0741: 5īpart of odc gene of Leishmania tarentolae for chromosomal integration nt 0742-0758: T7 promoter nt 0768-0784: TET operator nt 0820-1176: 5īnon-translated region of aprt gene of Leishmania tarentolae with splice acceptor site for target gene nt 1177-1196: BglII-NcoI-SlaI cloning sites nt 1186-1254: LMSAP signal peptide of L. mexicana secreted acid phosphatase nt 1246-1266: SalI-XbaI cloning sites nt 1267-2244: stuffer fragment to be replaced by target gene nt 2251-2262: MspCI-KpnI cloning sites nt 2263-2280: Hexa-His stretch coding region nt 2292-3631: 1.4k intergenic region from cam operon of Leishmania tarentolae with polyA site for target gene and splice acceptor site for marker gene nt 3641-4444: neo marker gene nt 4482-5059: 5'UTR of dhfr-ts gene of Leishmania with polyA site for marker gene nt 5183-5965: 3īpart of odc gene of Leishmania tarentolae for chromosomal integration nt 5967-7917: pUC18 derived with ori and bla marker gene @ nt 6250-7110. ATTTAAATGAATACTTACAAGGTACTCCTGCACCCGGCACAAGCGTATGTAATTGCTAGCACGCCGGTGCCGACCCAACCTCCAACTCCCGTGCTCACGCGCGCACACACGCACGCACAAGCATCATCATCATCATCATCATGGTCATCCCTCCCTTATCTGGCGAAGGAGGCACTGTGACGCAAGGTTTTCTGTGTCTCTCCTACCCCTCTACTGATTCCTTCTTTCTCTCTTCCTCTTCTCGCACTGCACCCCCCTCCCCCCCCCCACACACACACACTTGCTGTCATGGCTGTGCGCATTCGCTCACTCTTGTCGTGTCCCGTTTCCCTCATGGCGTCGTCGTCTTCCCCTGCGCAACCACCCACGCTGCACACGCACGCTCCACAGCGCGCTTCGCCTTCTTCGCTCTCTCCCTAGCCTCCTCTCTGTCACCCTCCACTGCCGCCGCGGTTGGCCGCTCTGCATCCTTTTCCCGCAGCCTTTCCTTTTTTTTTTCACTGTCAGTCTGGCTCATACCCGTGGAGCTGTGTCTCCCCGAGCTTCCCATACACCCACCCCCCACGACCTTTCTCAGCTTCCTTTCTCAGTCGCTTGAGCACGCAGTGCATTCACTTCTTCGTCCCTCAGTTCGCCTCGCTCTCGCTCTCTCTCGCGCGCGCTGCCGTACCCGCCCCTCCTCGTACTCCCCTCCTTCCTCATCCCCACCCCCCCCGTGTGCGCCGGCTCCTCTCTGAATTCTAATACGACTCACTATAGGGAGATCGtccctatcagtgatagaGATCCAGTCGCAGCCTGACCGCATCACACATCAAGgCGTTACAGCCTCTCCGTCTTTGCTGCACCCAAGtGTATTCGTGTGACCGCGTTGAGGGATCTAATGGATTCGGAACTTTGGTCGTTTGGTCTCTCATGCTGCCCAATCCCATGCGCTTTTCTCCGTGCCTCTCTGTCTCCCCTTCCGTTCTATTTGGACAACGATGTGTACTGTAATGcGTGCGCCATCTGACAACGAATAGATCAGCAgcATTCGCACACTTGCACATACCCAGTGAAGCTTTTGTGTCTGTCGTATTGACAACACCGACTGCAACAAGGTGTAGATAGAAGTTGGCCTTCTCGCTCGCTCGCACGCTCTTCACGCTCCTGCTTTCCTTGCTGTGCCTTgCCACCAgatctgCCATGGCcTCgAGGCTCGTCCGTGTGCTGGCcGCCGCCATGCTGGTTGCAGCGGCCGTGTCGGTCGACGCtGGCgCCtctctagacacaatatgtataggctaccatgcgaacaattcaaccgacactgttgacacagtgctcgagaagaatgtgacagtgacacactctgttaacctgctcgaagacagccacaacggaaaactatgtagattaaaaggaatagccccactacaattggggaaatgtaacatcgccggatggctcttgggaaacccagaatgcgacccactgcttccagtgagatcatggtActacattgtagaaacaccaaactctgagaatggaatatgttatccaggagatttcatcgactatgaggagctgagggagcaattgagctcagtgtcatcattcgaaagattcgaaatatttcccaaagaaagctcatggcccaaccacaacacaaAcaaaggagtaacggcagcatgctcccatgcggggGaaagcagtttttacagaaatttgctatggctgacggagaaggagggctcatacccaaagctgaaaaattcttatgtgaacaagaGagggaaagaagtccttgtactgtggggtattcatcacccgtctaacagtaaggaGcaacagaatatctatcagaatgaaaatgcttatgtctctgtagtgacttcaaattataacaggagatttaccccggaaatagcagaaagacccaaagtaagagatcaagctgggaggatgaactattactggaccttgctaaaacccggagacacaataatatttgaggcaaatggaaatctaatagcaccaaggtatgctttcgcactgagtagaggctttgggtccggcatcatcacctcaaacgcatcaatgcatgagtgtaacacgaagtgtcaaacacccctgggagctataaacagcagtctccctttccagaatatacacccagtcacaataggagagtgcccaaaatacgtcaggagtgccaaattgaggatggttacaggactaaggaacattccgtccattcaatccGCTAGCCTTAAGGGTACCCACCACCATCACCACCACTAGGCGGCCGCCCTCCTCCTCCTTTCTTGTTCCTTTCACGTCGCCTTCTCGGTTGTAGCTGGCAGACGACGAGTCTTACTTTTACGTGTACTTCTCTATAGATGATGTATGATCTCTCTGCATGCGTGTTCGTGCATGTGTCCGTGTGTTGTGTACGCGTGCGTCTCGCCTCAGCTCTCCGCGTGAAAGGGTTTGACTGCCCATGATGCGTGTGTATATCCACGCGCAGGCACGCACACACACACACAcaCACACACACACAggCACACACAGGCACACAAACGCATCTCAGGCCGAGCCGCATACGTCTCTCGCACGGTCTCGTTTATTTGATCATGTAGTTGAGTATTAAATTGGGAAGACAAAAACATAATAGCACGAAGAGTCGGGCACGAAAAGCCCGATCTCTCTCTCTCTCTCTCGTGCGCGCAGGGGCGTGTGGGTGCACGACGACGAGGACGGAGGGGGGAAGGGAGGCATAAACGgatcGATCCCCCATCGCACATGCGGGTACGAGCATTATCTGCTGTGTCTGGTCCTTTATCATATCGCTACCCGCCCGCCCCCCGCCCCCCTCCCCCCCCCCCCCCCcGcGGcTcCTGTCACTGTCGTTCCTGCGACTCCCCACACACCCGCGCAACGCTCATGTCACGAAGAGAAAAGTAGAAGGCGTGGCGATGCGTTGCGCGGCCCCCTGTCCGCTGGCATGCAGACGTCGAGCCGTGGAACGGATGGTAGCGAGAGACAACGCGGCGATGCAGGAAAGGAGATTTACTGCAGGACGTCTACACACGCGCGCACACCACTGCGAATGGCACCGTGCTGAGGAAAATTGGGGGGGAGGGCGCACGGGGGCGGGGCAGGAAACGGTTGGATCAGCAGCACTCTCAACTTGTGTCCGTATTGCGAAGGAGGGAGAGGAGCCTGCCCGTATTATGGGCGCTGAAACGTCGGAACCACCTATGAATCCCACTTCTGCGTCGCGGAGGCGTACTGCTTCCGACCACCTCCGCAGCTCACGCGCGTCGAGCTCCCTCCCTTTCCCCTTCTTTCCGTTGTGTGTGTGTGTGTGTGTGTGTCTTGTCACGATGCATGGCCATCTCTCCTCCAACCCTCCACGCCTTCCTCTCCCCCGCCCATCAAACGCGCTACGGCCACAACCATTTCTTCAAGTATCACATCCACCACCACTCTTACCCACCTACCCTCTTCGACTCTGACACAGCCACTGACGCCCTCTCCGCCTCTCTCGCTCGCTGCACATTGACTCTCCACAGCCCCCCCTCCCTCGCACAGCCAGCCccacccacCCACCCACCCACCCACCGCTCTACACACCTCCACAGCAGCCggatCCAATATGGGATCGGCCATTGAACAAGATGGATTGCACGCAGGTTCTCCGGCCGCTTGGGTGGAGAGGCTATTCGGCTATGACTGGGCACAACAGACAATCGGCTGCTCTGATGCCGCCGTGTTCCGGCTGTCAGCGCAGGGGCGCCCGGTTCTTTTTGTCAAGACCGACCTGTCCGGTGCCCTGAATGAACTGCAGGACGAGGCAGCGCGGCTATCGTGGCTGGCCACGACGGGCGTTCCTTGCGCAGCTGTGCTCGACGTTGTCACTGAAGCGGGAAGGGACTGGCTGCTATTGGGCGAAGTGCCGGGGCAGGATCTCCTGTCATCTCACCTTGCTCCTGCCGAGAAAGTATCCATCATGGCTGATGCAATGCGGCGGCTGCATACGCTTGATCCGGCTACCTGCCCATTCGACCACCAAGCGAAACATCGCATCGAGCGAGCACGTACTCGGATGGAAGCCGGTCTTGTCGATCAGGATGATCTGGACGAAGAGCATCAGGGGCTCGCGCCAGCCGAACTGTTCGCCAGGCTCAAGGCGCGCATGCCCGACGGCGAGGATCTCGTCGTGACgCATGGCGATGCCTGCTTGCCGAATATCATGGTGGAAAATGGCCGCTTTTCTGGATTCATCGACTGTGGCCGGCTGGGTGTGGCGGACCGCTATCAGGACATAGCGTTGGCTACCCGTGATATTGCTGAAGAGCTTGGCGGCGAATGGGCTGACCGCTTCCTCGTGCTTTACGGTATCGCCGCTCCCGATTCGCAGCGCATCGCCTTCTATCGCCTTCTTGACGAGTTCTTCTGAaCTAGTTCTAGTTCTAGGGGcCGAATTAATTCAGATCCTCGTGTGAGCGTTCGCGGAATCGGTCGCTCGTGTTTATGCCCGTCTTGGTGTTGTGCTCGCAAGGCGGTGCAGCAGGATACCGTCGCCCTCCTCTCTCCTTGCTTCTCTGTTCTTCAATTCGCGATCTCACAGAGGCCGGCTGTGCACGCCCTTCCTCACCCTCCTTTTCCCACCTCTCGGCCACCGGTCGGCTCCGTTCCGTCTGCCGTCGAGAAGGGACGGGCATGTGCAGCTCCTCCCTTTCTCTCGCGCGCGCATCTTCTCTTGCTTGTGGCACTCACGCTCATGCGTCAAGGCGGCCCCACGcCGAGCCCCTGCGCTCCCTTCCCTCTTGCGCATCCGTAGCgcGgaTGccGTCGATGCGCAAGGCCGGCATGAAGGAGCGCGTGCCCTCAAGAggGCacACTATCATGCCCTACGTGGGCCACGCAGCGATGAGGCCGGCTTCgGCGGAGATGCGTCACGCACGTGCCAGATGATGCgcTACGCCTTCCTTGACTTGCGCCCCCCTCTCTTCCTCCGTCTCTCACTCTCTCTCTCTCACACACACACACACACACACACACACACAAAGCTCCGGTTCGTCCGGGCCCTGCAGGCATGCCGACGTTCGGTGACCTGAACCACCTCGTCGCCGCTGTGATGTCTGGCGTGACCTGCTGCCTGCGCTTCCCTGGCCAGCTGAACTCTGACCTGCGCAAGCTTCTCCTGCTCAACGACGGCGACGGCGCTGACGAGGTGCAGAGTGGCACAGAAGCCGCAGCGGCGTACAGCGACGAGGAGGGCGGGAAGTCGCCGAGTGGTCCCCTGGCGAACGACTCCCTGTTCATGTCCGCGTGGGACCGCCGTCGCAACCTCACGAGCCGCCCGCTGCGCATCACAACCATATTCGGCCCCACCTGCGATTCGATGGACTGCATTCTGAAGAAGCAGCCCTTCCCGGAGATGAAGCTCGGCGACTGGCTTCTAGTGCCAGACATGGGCAGCTACACCACCGCTGCCGCGGGAGTCTTCAACGGTTTCGCGACACGCCGCCTTGAGTTTGTGAGCTCTGtTGACTTGTCTGCGAGGTCGAGGGCTGTGCATGCGCATGAGGGCAACACTTTTGAGTTcGTGAGCGAGTGAAAACGGGGCTGAGAGGATACAAGGTGCTGTAGGATGaGATCCTCCGCCCACCAGAGCATGGGCTATCTCTTGTGTCTGTTCTGAGCCTGCCTCTCTCTCTCTCTCTTGCCTCCCCCTCCCCCCTCCCCACACACACCCCACACGCACGCTCTGCGTCACCCTCTCCTCCCTCAGTAACGGGAGTAGACATGCAGAAGAAGTATACCACTAATACGTATGCGTTCCTGCAGGCATCCGAGATGGACACCAGAGCAAGCGCGGGTGTGCGCGAGTGGCCAGGAAAGGGGAAGAGAGAGAAAGAGAGAGAGACGAAGGAGCGCGTCCCGTCGTTGTGCGTCTTCTGTGGTGGCGCATTTAaATTTAGAGCTTGACGGGGAAAGCCGGCGAACGTGGCGAGAAAGGAAGGGAAGAAAGCGAAAGGAGCGGGCGCTAGGGCGCTGGCAAGTGTAGCGGTCACGCTGCGCGTAACCACCACACCCGcCGCGCTTAATGCGCCGCTACAGGGCGCGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCaTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTtCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTT