LOCUS pGL2316 6613 bp DNA circular UNA 01-APR-2009 DEFINITION plasmid for L. major ISP cluster KO PURr ACCESSION pGL1160 KEYWORDS 3` & 5` flanks; ampicillin resistance; hygromycin resistance; mexicana. SOURCE Expression vector pRIBOTEX Expression vector pRIBOTEX (japonica cultivar-group) ORGANISM Expression vector pRIBOTEX Expression vector pRIBOTEX artificial sequences; vectors FEATURES Location/Qualifiers Vector join(1624..6613,1..897) /origin="LoxP Construct C-terminal 6xHa (5,908bpcircular)Target vector: from KpnI cut at 918^919 to NdeIcut at 897^898, 5,887bpOrientation: originalPlus strand5': no changePlus strand 3': no changeMinus strand5': no changeMinus strand 3': no change" primer 537..552 /label="M13 F" misc_feature 675..708 /feature_type="LoxP Site" /label="LoxP" misc_feature 709..895 /feature_type="Splice Acceptor Site" /label="pX SL" misc_feature join(<1624..1630,896..>897) /feature_type="Insertion" /label="GOI Insertion Site" Site join(<898..>899,<900..901,897^898) /site_type="restriction site" /note="Name: NdeI" /note="Pattern: catatg" /note="cut_0_on_positive_strand: 6^7" /note="cut_0_on_negative_strand: 8^9" /note="inhibited_by: 5': N6-methyladenosine" /note="star_activity: yes" /note="site_type: other" /note="restriction site" /label="NdeI" primer <898..919 /label="Forward Primer" CDS 902..1618 /vntifkey="4" /label="GFP" /label="GFP from pGL1773" primer 1601..>1623 /label="Reverse Primer" Site join(1619..>1619,1620..>1623,1623^1624) /site_type="restriction site" /note="Name: KpnI" /note="Pattern: ggtacc" /note="cut_0_on_positive_strand: 735^736" /note="cut_0_on_negative_strand: 731^732" /note="inhibited_by: 5': N6-methyladenosine" /note="site_type: other" /note="restriction site" /label="KpnI" protein_bind 1637..1801 /label="6xHA" misc_feature 1808..1841 /feature_type="LoxP Site" /label="LoxP" polyA_site 1850..2345 primer complement(1988..2002) /label="OL2060" misc_signal 2346..2551 /label="Splice Acceptor Site" CDS 2552..3148 /label="PAC" Site join(<3162..>3165,<3166..3166,3161^3162) /site_type="restriction site" /note="Name: XhoI" /note="Pattern: ctcgag" /note="cut_0_on_positive_strand: 380^381" /note="cut_0_on_negative_strand: 384^385" /note="inhibited_by: 5': N6-methyladenosine" /note="site_type: other" /note="restriction site" /label="XhoI" primer <3162..3183 /label="OL4293 Forward Primer for Insertion into LoxP" misc_feature 3167..3354 /feature_type="Splice acceptor site" /label="SAS" primer complement(3337..3354) /label="OL4292 Mutagenesis Reverse Primer for XhoI Removal" primer 3355..3375 /label="OL4291 Mutagenesis Forward Primer for XhoI Removal" CDS 3355..3440 /vntifkey="4" /label="HASPB N-term" primer complement(3432..3449) /label="OL4290 Mutagenesis Reverse Primer for EF1a removal" primer 3450..3470 /label="OL4289 Mutagenesis Forward Primer for EF1a removal" CDS 3458..4165 /vntifkey="21" /label="mCherry" Tag 4166..4183 /label="His Tag" primer 4177..>4195 /label="OL4294 Reverse Primer for Insertion into LoxP" Site join(4194..>4195,complement(4196..>4199),4195^4196,complement(4195^4196)) /site_type="restriction site" /note="Name: NotI" /note="Pattern: gcggccgc" /note="cut_0_on_positive_strand: 1516^1517" /note="cut_0_on_negative_strand: 1520^1521" /note="inhibited_by: 5': N4-methylcytosine" /note="site_type: other" /note="restriction site" /label="NotI" polyA_site 4205..4729 /label="Lmcpb Poly A" polyA_signal 4632..4646 /label="S1" primer complement(4878..4894) /label="M13 R" ORIGIN 1 CTTTCCTGCG TTATCCCCTG ATTCTGTGGA TAACCGTATT ACCGCCTTTG AGTGAGCTGA 61 TACCGCTCGC CGCAGCCGAA CGACCGAGCG CAGCGAGTCA GTGAGCGAGG AAGCGGAAGA 121 GCGCCCAATA CGCAAACCGC CTCTCCCCGC GCGTTGGCCG ATTCATTAAT GCAGCTGGCA 181 CGACAGGTTT CCCGACTGGA AAGCGGGCAG TGAGCGCAAC GCAATTAATA CGCGTACCGC 241 TAGCCAGGAA GAGTTTGTAG AAACGCAAAA AGGCCATCCG TCAGGATGGC CTTCTGCTTA 301 GTTTGATGCC TGGCAGTTTA TGGCGGGCGT CCTGCCCGCC ACCCTCCGGG CCGTTGCTTC 361 ACAACGTTCA AATCCGCTCC CGGCGGATTT GTCCTACTCA GGAGAGCGTT CACCGACAAA 421 CAACAGATAA AACGAAAGGC CCAGTCTTCC GACTGAGCCT TTCGTTTTAT TTGATGCCTG 481 GCAGTTCCCT ACTCTCGCGT TAACGCTAGC ATGGATGTTT TCCCAGTCAC GACGTTGTAA 541 AACGACGGCC AGTCTTAAGC TCGGGCCCCA AATAATGATT TTATTTTGAC TGATAGTGAC 601 CTGTTCGTTG CAACAAATTG ATGAGCAATG CTTTTTTATA ATGCCAACTT TGTACAAAAA 661 AGCAGGCTTA GATCATAACT TCGTATAGCA TACATTATAC GAAGTTATAA TTGCCCGCTT 721 TCCATTTCGT CACCTTCCGC CTCTCTCTCT CTCTCTCTCA CCATCTACGC GTGCACATCA 781 TCAACTGTCT CTTGTCGGTG CTCACCACCC TCAACCACCC CTCACTTTCA AGGCTTCCCG 841 AACGCACACA AAAGGCGTGA AAACCGCTCG CGTGTGTTGA GCCGTCCACC GTAGCCATAT 901 GATGGTGAGC AAGGGCGAGG AGCTGTTCAC CGGGGTGGTG CCCATCCTGG TCGAGCTGGA 961 CGGCGACGTA AACGGCCACA AGTTCAGCGT GTCCGGCGAG GGCGAGGGCG ATGCCACCTA 1021 CGGCAAGCTG ACCCTGAAGT TCATCTGCAC CACCGGCAAG CTGCCCGTGC CCTGGCCCAC 1081 CCTCGTGACC ACCCTGACCT ACGGCGTGCA GTGCTTCAGC CGCTACCCCG ACCACATGAA 1141 GCAGCACGAC TTCTTCAAGT CCGCCATGCC CGAAGGCTAC GTCCAGGAGC GCACCATCTT 1201 CTTCAAGGAC GACGGCAACT ACAAGACCCG CGCCGAGGTG AAGTTCGAGG GCGACACCCT 1261 GGTGAACCGC ATCGAGCTGA AGGGCATCGA CTTCAAGGAG GACGGCAACA TCCTGGGGCA 1321 CAAGCTGGAG TACAACTACA ACAGCCACAA CGTCTATATC ATGGCCGACA AGCAGAAGAA 1381 CGGCATCAAG GTGAACTTCA AGATCCGCCA CAACATCGAG GACGGCAGCG TGCAGCTCGC 1441 CGACCACTAC CAGCAGAACA CCCCCATCGG CGACGGCCCC GTGCTGCTGC CCGACAACCA 1501 CTACCTGAGC ACCCAGTCCG CCCTGAGCAA AGACCCCAAC GAGAAGCGCG ATCACATGGT 1561 CCTGCTGGAG TTCGTGACCG CCGCCGGGAT CACTCTCGGC ATGGACGAGC TGTACAAGGG 1621 TACCATCGAT ACTAGTTACC CTTACGATGT GCCTGATTAC GCGTACCCAT ACGACGTGCC 1681 AGACTACGCA TACCCGTACG ATGTGCCCGA TTACGCATAC CCTTACGATG TGCCTGATTA 1741 CGCGTACCCA TACGACGTGC CAGACTACGC ATACCCGTAC GATGTGCCCG ATTACGCATG 1801 ATCTAGAATA ACTTCGTATA GCATACATTA TACGAAGTTA TGGCTGCAGC GCGTGGATGT 1861 CGCGCAGCGC AAGACCCCGT GCGCCTGAAA CGTGAAGGGA GGTCGAGAAG CGTGCCGCAT 1921 GAGGCCTTAA GGCAGGAAAG TGAAAAGAGC TCGACGCGAG TGCAGCACGC GCGACATCGA 1981 ACAGGACGGA ATAGACACTC AGCCCCTTCC CCTTTCAAAA ACTGAATGGA CGGACATCGT 2041 AACGCGCTTC TCTCCCCTCC ACTCCCCCTG TCCCTGTCTC CTTGACGTGT GCTGCTCTCG 2101 TGAGCGACAG ATGAAGCGAG GCGTAGAAGG AAAGGGGGGG GAGCGAGGCC TGCACACACA 2161 CACACACACA CACACACACA CACACACACA CACACACACA CACACACACG AAAAGCAAGT 2221 TTGAGGCAGA CGTGGTTTCA CACGATTCCT GTGCAAGAAG CGTGCCTGCG TGACGGTGTG 2281 GCTGTGTATG CATGCGTGAA GGGTTACCGC AACGATGCGC AATGGACTCC CCCGCTTTCC 2341 ATTTCGTCAC CTTCCGCCTC TCTCTCTCTC TCTCTCACCA TCTACGCGTG CACATCATCA 2401 ACTGTCTCTT GTCGGTGCTC ACCACCCTCA ACCACCCCTC ACTTTCAAGG CTTCCCGAAC 2461 GCACACAAAA GGCGTGAAAA CCGCTCGCGT GTGTTGAGCC GTCCACCGTA GCCCTCCCCC 2521 TGTCCCCGGG GGATCCACTA GACGACCTTC CATGACCGAG TACAAGCCCA CGGTGCGCCT 2581 CGCCACCCGC GACGACGTCC CCCGGGCCGT ACGCACCCTC GCCGCCGCGT TCGCCGACTA 2641 CCCCGCCACG CGCCACACCG TCGACCCGGA CCGCCACATC GAGCGGGTCA CCGAGCTGCA 2701 AGAACTCTTC CTCACGCGCG TCGGGCTCGA CATCGGCAAG GTGTGGGTCG CGGACGACGG 2761 CGCCGCGGTG GCGGTCTGGA CCACGCCGGA GAGCGTCGAA GCGGGGGCGG TGTTCGCCGA 2821 GATCGGCCCG CGCATGGCCG AGTTGAGCGG TTCCCGGCTG GCCGCGCAGC AACAGATGGA 2881 AGGCCTCCTG GCGCCGCACC GGCCCAAGGA GCCCGCGTGG TTCCTGGCCA CCGTCGGCGT 2941 CTCGCCCGAC CACCAGGGCA AGGGTCTGGG CAGCGCCGTC GTGCTCCCCG GAGTGGAGGC 3001 GGCCGAGCGC GCCGGGGTGC CCGCCTTCCT GGAGACCTCC GCGCCCCGCA ACCTCCCCTT 3061 CTACGAGCGG CTCGGCTTCA CCGTCACCGC CGACGTCGAG TGCCCGAAGG ACCGCGCGAC 3121 CTGGTGCATG ACCCGCAAGC CCGGTGCCTA GGCTAGCGCT CTCGAGAATT GCCCGCTTTC 3181 CATTTCGTCA CCTTCCGCCT CTCTCTCTCT CTCTCTCACC ATCTACGCGT GCACATCATC 3241 AACTGTCTCT TGTCGGTGCT CACCACCCTC AACCACCCCT CACTTTCAAG GCTTCCCGAA 3301 CGCACACAAA AGGCGTGAAA ACCGCTCGCG GTGTGTTGAG CCGTCCACCG TAGCTGATCC 3361 TATCTATCTC CCCCGCTTAT ACACCATGGG AAGCTCTTGC ACGAAGGACT CCGCAAAGGA 3421 GCCCCAGAAA AGTGCGGATG GATCTATGGA GGATCCAATG GTTAGTAAAG GAGAAGAAAA 3481 TAACATGGCA ATCATTAAGG AGTTCATGAG ATTCAAAGTT CACATGGAAG GTTCTGTAAA 3541 TGGACATGAA TTTGAAATAG AAGGTGAAGG AGAAGGAAGG CCTTATGAAG GAACCCAAAC 3601 CGCGAAGCTA AAAGTTACTA AGGGTGGCCC ATTACCATTT GCATGGGATA TCCTTAGCCC 3661 TCAATTCATG TATGGGTCAA AGGCTTATGT CAAGCACCCC GCCGACATTC CAGACTATCT 3721 AAAGTTATCT TTTCCCGAAG GGTTTAAGTG GGAGCGTGTG ATGAACTTCG AAGACGGTGG 3781 CGTGGTAACA GTGACTCAGG ATTCGTCCCT GCAAGATGGT GAATTTATCT ACAAAGTCAA 3841 ATTAAGAGGA ACTAACTTTC CATCTGGCGG CCCGGTTATG CAAAAAAAGA CAATGGGCTG 3901 GGAGGCCTCC TCAGAACGAA TGTACCCTGA AGATGGTGCC TTGAAGGGTG AGATTAAACA 3961 AAGATTGAAA TTGAAAGATG GTGGACATTA TGACGCTGAG GTTAAAACGA CATACAAAGC 4021 TAAGAAACCT GTCCAGCTCC CAGGTGCTTA CAATGTAAAT ATAAAACTTG ATATTACATC 4081 ACATAATGAA GATTATACGA TAGTTGAACA ATACGAAAGG GCTGAGGGGA GACATAGTAC 4141 TGGTGGCATG GATGAACTAT ACAAACATCA TCATCATCAT CATTGAGGAT CCCGCGGCCG 4201 CTAGATGTGT GCCCTTGTGT GCGTGTGTGT TTGGTGGTTT GCAGCGATGC CCGGCGCGTG 4261 TGGGCACCTC CTTGGGTGCG CGCCCGCCGT GGCAGCTGCG CGTGCGTGCG AGATGTGAGG 4321 CAGAGGAAGA GGAAGGCGAT GCGGGCGACA GCGCAGCGAG GTGCGGCGGA GCGTAGGGGG 4381 GAAATGGACG AGCAGGCGCG CTGTGAATCG GAGCTGCGGC ACCACCCAAG TCGTGGTGCC 4441 CCGCGAATGG CTGTTCTGCC CGCCCTCGCT TCACGCCTCC CCCTCCCCTC GCGTGCCCTC 4501 GCGTGGCCTC CCTTGTTATC CCTCTCTCTC GCACGCACAC GCACACGCGT ATACGCGAGC 4561 CCGCTATTCT GCCTTCGTCT GGCTCTTTGT ATTTTGCTTG CTTCTTCAGC ACACTTGTGT 4621 GCTGTGCGTT CAGCGATATC TTCCACTACT TTGTTTTCTC CTCCCCCTCG GGAGGTGCTT 4681 CGCTTGTGCT TTGACGGTGG TGCGTGGCTG CTGGGTTATG TGCCGGGCGC CCATGGAACC 4741 CAGCTTTCTT GTACAAAGTT GGCATTATAA GAAAGCATTG CTTATCAATT TGTTGCAACG 4801 AACAGGTCAC TATCAGTCAA AATAAAATCA TTATTTGCCA TCCAGCTGAT ATCCCCTATA 4861 GTGAGTCGTA TTACATGGTC ATAGCTGTTT CCTGGCAGCT CTGGCCCGTG TCTCAAAATC 4921 TCTGATGTTA CATTGCACAA GATAAAAATA TATCATCATG AACAATAAAA CTGTCTGCTT 4981 ACATAAACAG TAATACAAGG GGTGTTATGA GCCATATTCA ACGGGAAACG TCGAGGCCGC 5041 GATTAAATTC CAACATGGAT GCTGATTTAT ATGGGTATAA ATGGGCTCGC GATAATGTCG 5101 GGCAATCAGG TGCGACAATC TATCGCTTGT ATGGGAAGCC CGATGCGCCA GAGTTGTTTC 5161 TGAAACATGG CAAAGGTAGC GTTGCCAATG ATGTTACAGA TGAGATGGTC AGACTAAACT 5221 GGCTGACGGA ATTTATGCCT CTTCCGACCA TCAAGCATTT TATCCGTACT CCTGATGATG 5281 CATGGTTACT CACCACTGCG ATCCCCGGAA AAACAGCATT CCAGGTATTA GAAGAATATC 5341 CTGATTCAGG TGAAAATATT GTTGATGCGC TGGCAGTGTT CCTGCGCCGG TTGCATTCGA 5401 TTCCTGTTTG TAATTGTCCT TTTAACAGCG ATCGCGTATT TCGTCTCGCT CAGGCGCAAT 5461 CACGAATGAA TAACGGTTTG GTTGATGCGA GTGATTTTGA TGACGAGCGT AATGGCTGGC 5521 CTGTTGAACA AGTCTGGAAA GAAATGCATA AACTTTTGCC ATTCTCACCG GATTCAGTCG 5581 TCACTCATGG TGATTTCTCA CTTGATAACC TTATTTTTGA CGAGGGGAAA TTAATAGGTT 5641 GTATTGATGT TGGACGAGTC GGAATCGCAG ACCGATACCA GGATCTTGCC ATCCTATGGA 5701 ACTGCCTCGG TGAGTTTTCT CCTTCATTAC AGAAACGGCT TTTTCAAAAA TATGGTATTG 5761 ATAATCCTGA TATGAATAAA TTGCAGTTTC ATTTGATGCT CGATGAGTTT TTCTAATCAG 5821 AATTGGTTAA TTGGTTGTAA CACTGGCAGA GCATTACGCT GACTTGACGG GACGGCGCAA 5881 GCTCATGACC AAAATCCCTT AACGTGAGTT ACGCGTCGTT CCACTGAGCG TCAGACCCCG 5941 TAGAAAAGAT CAAAGGATCT TCTTGAGATC CTTTTTTTCT GCGCGTAATC TGCTGCTTGC 6001 AAACAAAAAA ACCACCGCTA CCAGCGGTGG TTTGTTTGCC GGATCAAGAG CTACCAACTC 6061 TTTTTCCGAA GGTAACTGGC TTCAGCAGAG CGCAGATACC AAATACTGTT CTTCTAGTGT 6121 AGCCGTAGTT AGGCCACCAC TTCAAGAACT CTGTAGCACC GCCTACATAC CTCGCTCTGC 6181 TAATCCTGTT ACCAGTGGCT GCTGCCAGTG GCGATAAGTC GTGTCTTACC GGGTTGGACT 6241 CAAGACGATA GTTACCGGAT AAGGCGCAGC GGTCGGGCTG AACGGGGGGT TCGTGCACAC 6301 AGCCCAGCTT GGAGCGAACG ACCTACACCG AACTGAGATA CCTACAGCGT GAGCTATGAG 6361 AAAGCGCCAC GCTTCCCGAA GGGAGAAAGG CGGACAGGTA TCCGGTAAGC GGCAGGGTCG 6421 GAACAGGAGA GCGCACGAGG GAGCTTCCAG GGGGAAACGC CTGGTATCTT TATAGTCCTG 6481 TCGGGTTTCG CCACCTCTGA CTTGAGCGTC GATTTTTGTG ATGCTCGTCA GGGGGGCGGA 6541 GCCTATGGAA AAACGCCAGC AACGCGGCCT TTTTACGGTT CCTGGCCTTT TGCTGGCCTT 6601 TTGCTCACAT GTT //