LOCUS pGL2315 6463 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(1653..6463,1..926) /origin="LoxP Construct C-terminal 6xHa (5,908bpcircular)Target vector: from XbaI cut at 1097^1098 toSpeI cut at 926^927, 5,737bpOrientation: originalPlusstrand 5': no changePlus strand 3': no changeMinusstrand 5': no changeMinus strand 3': no change" primer 537..552 /label="M13 F" primer complement(655..674) /label="OL4489" misc_feature 675..708 /feature_type="LoxP Site" /label="LoxP" primer 703..722 /label="OL4611" misc_feature 709..895 /feature_type="Splice Acceptor Site" /label="pX SL" misc_feature 896..925 /feature_type="Insertion" /label="GOI Insertion Site" Site join(<927..>930,<931..931,926^927) /site_type="restriction site" /note="Name: SpeI" /note="Pattern: actagt" /note="cut_0_on_positive_strand: 1^2" /note="cut_0_on_negative_strand: 5^6" /note="inhibited_by: " /note="star_activity: yes" /note="site_type: other" /note="restriction site" /label="SpeI" primer <927..949 /label="Forward Primer" CDS 932..1648 /vntifkey="4" /label="GFP" /label="GFP from pGL1773" primer 1631..>1652 /label="Reverse Primer" Site join(1652..>1652,complement(1653..>1656),1652^1653,complement(1652^1653)) /site_type="restriction site" /note="Name: XbaI" /note="Pattern: tctaga" /note="cut_0_on_positive_strand: 751^752" /note="cut_0_on_negative_strand: 755^756" /note="inhibited_by: 5': N6-methyladenosine" /note="star_activity: yes" /note="site_type: other" /note="restriction site" /label="XbaI" misc_feature 1658..1691 /feature_type="LoxP Site" /label="LoxP" polyA_site 1700..2195 primer complement(1838..1852) /label="OL2060" misc_signal 2196..2401 /label="Splice Acceptor Site" CDS 2402..2998 /label="PAC" Site join(<3012..>3015,<3016..3016,3011^3012) /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 <3012..3033 /label="OL4293 Forward Primer for Insertion into LoxP" misc_feature 3017..3204 /feature_type="Splice acceptor site" /label="SAS" primer complement(3187..3204) /label="OL4292 Mutagenesis Reverse Primer for XhoI Removal" CDS 3205..3290 /vntifkey="4" /label="HASPB N-term" primer 3205..3225 /label="OL4291 Mutagenesis Forward Primer for XhoI Removal" primer complement(3282..3299) /label="OL4290 Mutagenesis Reverse Primer for EF1a removal" primer 3300..3320 /label="OL4289 Mutagenesis Forward Primer for EF1a removal" CDS 3308..4015 /vntifkey="21" /label="mCherry" Tag 4016..4033 /label="His Tag" primer 4027..>4045 /label="OL4294 Reverse Primer for Insertion into LoxP" Site join(4044..>4045,complement(4046..>4049),4045^4046,complement(4045^4046)) /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 4055..4579 /label="Lmcpb Poly A" polyA_signal 4482..4496 /label="S1" primer complement(4728..4744) /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 GGAATTCAGT ACTGGTACCA TCGATACTAG TATGGTGAGC AAGGGCGAGG AGCTGTTCAC 961 CGGGGTGGTG CCCATCCTGG TCGAGCTGGA CGGCGACGTA AACGGCCACA AGTTCAGCGT 1021 GTCCGGCGAG GGCGAGGGCG ATGCCACCTA CGGCAAGCTG ACCCTGAAGT TCATCTGCAC 1081 CACCGGCAAG CTGCCCGTGC CCTGGCCCAC CCTCGTGACC ACCCTGACCT ACGGCGTGCA 1141 GTGCTTCAGC CGCTACCCCG ACCACATGAA GCAGCACGAC TTCTTCAAGT CCGCCATGCC 1201 CGAAGGCTAC GTCCAGGAGC GCACCATCTT CTTCAAGGAC GACGGCAACT ACAAGACCCG 1261 CGCCGAGGTG AAGTTCGAGG GCGACACCCT GGTGAACCGC ATCGAGCTGA AGGGCATCGA 1321 CTTCAAGGAG GACGGCAACA TCCTGGGGCA CAAGCTGGAG TACAACTACA ACAGCCACAA 1381 CGTCTATATC ATGGCCGACA AGCAGAAGAA CGGCATCAAG GTGAACTTCA AGATCCGCCA 1441 CAACATCGAG GACGGCAGCG TGCAGCTCGC CGACCACTAC CAGCAGAACA CCCCCATCGG 1501 CGACGGCCCC GTGCTGCTGC CCGACAACCA CTACCTGAGC ACCCAGTCCG CCCTGAGCAA 1561 AGACCCCAAC GAGAAGCGCG ATCACATGGT CCTGCTGGAG TTCGTGACCG CCGCCGGGAT 1621 CACTCTCGGC ATGGACGAGC TGTACAAGTG ATCTAGAATA ACTTCGTATA GCATACATTA 1681 TACGAAGTTA TGGCTGCAGC GCGTGGATGT CGCGCAGCGC AAGACCCCGT GCGCCTGAAA 1741 CGTGAAGGGA GGTCGAGAAG CGTGCCGCAT GAGGCCTTAA GGCAGGAAAG TGAAAAGAGC 1801 TCGACGCGAG TGCAGCACGC GCGACATCGA ACAGGACGGA ATAGACACTC AGCCCCTTCC 1861 CCTTTCAAAA ACTGAATGGA CGGACATCGT AACGCGCTTC TCTCCCCTCC ACTCCCCCTG 1921 TCCCTGTCTC CTTGACGTGT GCTGCTCTCG TGAGCGACAG ATGAAGCGAG GCGTAGAAGG 1981 AAAGGGGGGG GAGCGAGGCC TGCACACACA CACACACACA CACACACACA CACACACACA 2041 CACACACACA CACACACACG AAAAGCAAGT TTGAGGCAGA CGTGGTTTCA CACGATTCCT 2101 GTGCAAGAAG CGTGCCTGCG TGACGGTGTG GCTGTGTATG CATGCGTGAA GGGTTACCGC 2161 AACGATGCGC AATGGACTCC CCCGCTTTCC ATTTCGTCAC CTTCCGCCTC TCTCTCTCTC 2221 TCTCTCACCA TCTACGCGTG CACATCATCA ACTGTCTCTT GTCGGTGCTC ACCACCCTCA 2281 ACCACCCCTC ACTTTCAAGG CTTCCCGAAC GCACACAAAA GGCGTGAAAA CCGCTCGCGT 2341 GTGTTGAGCC GTCCACCGTA GCCCTCCCCC TGTCCCCGGG GGATCCACTA GACGACCTTC 2401 CATGACCGAG TACAAGCCCA CGGTGCGCCT CGCCACCCGC GACGACGTCC CCCGGGCCGT 2461 ACGCACCCTC GCCGCCGCGT TCGCCGACTA CCCCGCCACG CGCCACACCG TCGACCCGGA 2521 CCGCCACATC GAGCGGGTCA CCGAGCTGCA AGAACTCTTC CTCACGCGCG TCGGGCTCGA 2581 CATCGGCAAG GTGTGGGTCG CGGACGACGG CGCCGCGGTG GCGGTCTGGA CCACGCCGGA 2641 GAGCGTCGAA GCGGGGGCGG TGTTCGCCGA GATCGGCCCG CGCATGGCCG AGTTGAGCGG 2701 TTCCCGGCTG GCCGCGCAGC AACAGATGGA AGGCCTCCTG GCGCCGCACC GGCCCAAGGA 2761 GCCCGCGTGG TTCCTGGCCA CCGTCGGCGT CTCGCCCGAC CACCAGGGCA AGGGTCTGGG 2821 CAGCGCCGTC GTGCTCCCCG GAGTGGAGGC GGCCGAGCGC GCCGGGGTGC CCGCCTTCCT 2881 GGAGACCTCC GCGCCCCGCA ACCTCCCCTT CTACGAGCGG CTCGGCTTCA CCGTCACCGC 2941 CGACGTCGAG TGCCCGAAGG ACCGCGCGAC CTGGTGCATG ACCCGCAAGC CCGGTGCCTA 3001 GGCTAGCGCT CTCGAGAATT GCCCGCTTTC CATTTCGTCA CCTTCCGCCT CTCTCTCTCT 3061 CTCTCTCACC ATCTACGCGT GCACATCATC AACTGTCTCT TGTCGGTGCT CACCACCCTC 3121 AACCACCCCT CACTTTCAAG GCTTCCCGAA CGCACACAAA AGGCGTGAAA ACCGCTCGCG 3181 GTGTGTTGAG CCGTCCACCG TAGCTGATCC TATCTATCTC CCCCGCTTAT ACACCATGGG 3241 AAGCTCTTGC ACGAAGGACT CCGCAAAGGA GCCCCAGAAA AGTGCGGATG GATCTATGGA 3301 GGATCCAATG GTTAGTAAAG GAGAAGAAAA TAACATGGCA ATCATTAAGG AGTTCATGAG 3361 ATTCAAAGTT CACATGGAAG GTTCTGTAAA TGGACATGAA TTTGAAATAG AAGGTGAAGG 3421 AGAAGGAAGG CCTTATGAAG GAACCCAAAC CGCGAAGCTA AAAGTTACTA AGGGTGGCCC 3481 ATTACCATTT GCATGGGATA TCCTTAGCCC TCAATTCATG TATGGGTCAA AGGCTTATGT 3541 CAAGCACCCC GCCGACATTC CAGACTATCT AAAGTTATCT TTTCCCGAAG GGTTTAAGTG 3601 GGAGCGTGTG ATGAACTTCG AAGACGGTGG CGTGGTAACA GTGACTCAGG ATTCGTCCCT 3661 GCAAGATGGT GAATTTATCT ACAAAGTCAA ATTAAGAGGA ACTAACTTTC CATCTGGCGG 3721 CCCGGTTATG CAAAAAAAGA CAATGGGCTG GGAGGCCTCC TCAGAACGAA TGTACCCTGA 3781 AGATGGTGCC TTGAAGGGTG AGATTAAACA AAGATTGAAA TTGAAAGATG GTGGACATTA 3841 TGACGCTGAG GTTAAAACGA CATACAAAGC TAAGAAACCT GTCCAGCTCC CAGGTGCTTA 3901 CAATGTAAAT ATAAAACTTG ATATTACATC ACATAATGAA GATTATACGA TAGTTGAACA 3961 ATACGAAAGG GCTGAGGGGA GACATAGTAC TGGTGGCATG GATGAACTAT ACAAACATCA 4021 TCATCATCAT CATTGAGGAT CCCGCGGCCG CTAGATGTGT GCCCTTGTGT GCGTGTGTGT 4081 TTGGTGGTTT GCAGCGATGC CCGGCGCGTG TGGGCACCTC CTTGGGTGCG CGCCCGCCGT 4141 GGCAGCTGCG CGTGCGTGCG AGATGTGAGG CAGAGGAAGA GGAAGGCGAT GCGGGCGACA 4201 GCGCAGCGAG GTGCGGCGGA GCGTAGGGGG GAAATGGACG AGCAGGCGCG CTGTGAATCG 4261 GAGCTGCGGC ACCACCCAAG TCGTGGTGCC CCGCGAATGG CTGTTCTGCC CGCCCTCGCT 4321 TCACGCCTCC CCCTCCCCTC GCGTGCCCTC GCGTGGCCTC CCTTGTTATC CCTCTCTCTC 4381 GCACGCACAC GCACACGCGT ATACGCGAGC CCGCTATTCT GCCTTCGTCT GGCTCTTTGT 4441 ATTTTGCTTG CTTCTTCAGC ACACTTGTGT GCTGTGCGTT CAGCGATATC TTCCACTACT 4501 TTGTTTTCTC CTCCCCCTCG GGAGGTGCTT CGCTTGTGCT TTGACGGTGG TGCGTGGCTG 4561 CTGGGTTATG TGCCGGGCGC CCATGGAACC CAGCTTTCTT GTACAAAGTT GGCATTATAA 4621 GAAAGCATTG CTTATCAATT TGTTGCAACG AACAGGTCAC TATCAGTCAA AATAAAATCA 4681 TTATTTGCCA TCCAGCTGAT ATCCCCTATA GTGAGTCGTA TTACATGGTC ATAGCTGTTT 4741 CCTGGCAGCT CTGGCCCGTG TCTCAAAATC TCTGATGTTA CATTGCACAA GATAAAAATA 4801 TATCATCATG AACAATAAAA CTGTCTGCTT ACATAAACAG TAATACAAGG GGTGTTATGA 4861 GCCATATTCA ACGGGAAACG TCGAGGCCGC GATTAAATTC CAACATGGAT GCTGATTTAT 4921 ATGGGTATAA ATGGGCTCGC GATAATGTCG GGCAATCAGG TGCGACAATC TATCGCTTGT 4981 ATGGGAAGCC CGATGCGCCA GAGTTGTTTC TGAAACATGG CAAAGGTAGC GTTGCCAATG 5041 ATGTTACAGA TGAGATGGTC AGACTAAACT GGCTGACGGA ATTTATGCCT CTTCCGACCA 5101 TCAAGCATTT TATCCGTACT CCTGATGATG CATGGTTACT CACCACTGCG ATCCCCGGAA 5161 AAACAGCATT CCAGGTATTA GAAGAATATC CTGATTCAGG TGAAAATATT GTTGATGCGC 5221 TGGCAGTGTT CCTGCGCCGG TTGCATTCGA TTCCTGTTTG TAATTGTCCT TTTAACAGCG 5281 ATCGCGTATT TCGTCTCGCT CAGGCGCAAT CACGAATGAA TAACGGTTTG GTTGATGCGA 5341 GTGATTTTGA TGACGAGCGT AATGGCTGGC CTGTTGAACA AGTCTGGAAA GAAATGCATA 5401 AACTTTTGCC ATTCTCACCG GATTCAGTCG TCACTCATGG TGATTTCTCA CTTGATAACC 5461 TTATTTTTGA CGAGGGGAAA TTAATAGGTT GTATTGATGT TGGACGAGTC GGAATCGCAG 5521 ACCGATACCA GGATCTTGCC ATCCTATGGA ACTGCCTCGG TGAGTTTTCT CCTTCATTAC 5581 AGAAACGGCT TTTTCAAAAA TATGGTATTG ATAATCCTGA TATGAATAAA TTGCAGTTTC 5641 ATTTGATGCT CGATGAGTTT TTCTAATCAG AATTGGTTAA TTGGTTGTAA CACTGGCAGA 5701 GCATTACGCT GACTTGACGG GACGGCGCAA GCTCATGACC AAAATCCCTT AACGTGAGTT 5761 ACGCGTCGTT CCACTGAGCG TCAGACCCCG TAGAAAAGAT CAAAGGATCT TCTTGAGATC 5821 CTTTTTTTCT GCGCGTAATC TGCTGCTTGC AAACAAAAAA ACCACCGCTA CCAGCGGTGG 5881 TTTGTTTGCC GGATCAAGAG CTACCAACTC TTTTTCCGAA GGTAACTGGC TTCAGCAGAG 5941 CGCAGATACC AAATACTGTT CTTCTAGTGT AGCCGTAGTT AGGCCACCAC TTCAAGAACT 6001 CTGTAGCACC GCCTACATAC CTCGCTCTGC TAATCCTGTT ACCAGTGGCT GCTGCCAGTG 6061 GCGATAAGTC GTGTCTTACC GGGTTGGACT CAAGACGATA GTTACCGGAT AAGGCGCAGC 6121 GGTCGGGCTG AACGGGGGGT TCGTGCACAC AGCCCAGCTT GGAGCGAACG ACCTACACCG 6181 AACTGAGATA CCTACAGCGT GAGCTATGAG AAAGCGCCAC GCTTCCCGAA GGGAGAAAGG 6241 CGGACAGGTA TCCGGTAAGC GGCAGGGTCG GAACAGGAGA GCGCACGAGG GAGCTTCCAG 6301 GGGGAAACGC CTGGTATCTT TATAGTCCTG TCGGGTTTCG CCACCTCTGA CTTGAGCGTC 6361 GATTTTTGTG ATGCTCGTCA GGGGGGCGGA GCCTATGGAA AAACGCCAGC AACGCGGCCT 6421 TTTTACGGTT CCTGGCCTTT TGCTGGCCTT TTGCTCACAT GTT //