pXG-/GFP+ (aka pXG-‘GFP+) 11/01
strain B2863 Steve Beverley
Leishmania expression vector for making GFP fusions to C-terminal of your favorite protein.
B2863 pXG-/GFP+
Blue bars represent ORFs, red arrows indicated mRNAs known or predicted from the L. major intergenic regions. All derive from regions surrounding the DHFR-TS gene.
B2863 pXG-/GFP+
Gene fusions may be made conveniently by inserting your gene in the appropriate reading frame in the polylinker located 5’-ward of the GFP+ gene. The GFP+ gene contains GC-rich codons as well as mutations that improve the fluorescence signal (T65), making it suitable for ‘fluorescein’-oriented fluorescence optics.
Reference: for the parent pXG-GFP+ vector, Ha, D.S., Schwarz, J.K., S.J. Turco, and S.M. Beverley (1996), "Use of the green fluorescent protein as a marker in transfected Leishmania ", Mol. Biochem. Parasitol. 77: 57-64.
This construct has been used in several works in progress and was made by Haiyan Xu in our lab.
Polylinker in front of /GFP+ cassette:
SmaI BamHI SfiI EcoRV
CCC GGG GGA TCC GGC CGT GCC GGC CGA GAT ATC GTG AGC AAG GGC GAG GAG CTG TTC
Val Ser Lys Gly Glu Glu Leu Phe->
Sequence information for this construct is provided below: please contact Steve Beverley () if you find discrepancies.
LOCUS pXG-/GFP+ 7570 bp DNA CIRCULAR SYN 29-NOV-2001
DEFINITION vector for putting GFP at C terminus of your gene; pXG-'GFP+ alternate name
JOURNAL Unpublished.
FEATURES Location/Qualifiers
CDS 1288..1363
/gene="5'DST"
/product="5'DST"
misc_feature 1396..2118
/gene="'GFP+"
/product="GC rich GFP with S65T mutation, lacking ATG"
CDS 2164..2280
/gene="6.2-l"
/product="6.2-l"
CDS 2750..2902
/gene="1.7-l"
/product="1.7-l"
misc_feature complement (2932)
/gene="T3 promoter"
/product="T3"
misc_feature complement (3932..4692)
/gene="amp"
/product="ampicillin resistance"
misc_feature 5656
/gene="SP6"
/product="SP6 promoter"
CDS 5720..5919
/gene="3'13K"
/product="3'13K"
CDS 6414..879
/gene="NEO/DHFR_TS"
/product="NEO/DHFR-TS"
misc_feature 6707..7510
/gene="NEO"
BASE COUNT 1626 a 2262 c 2038 g 1644 t
ORIGIN
1 ACTAGTGATC CCGCCCGCAC CCGCGCGCGC TCAGGCCGCG TGTCGTCGTG CTCTTTCCAT
61 TTTTTTTTTG TGTGTGGTGG TGATTTGAGC TGCTCTCCGT TGTGTGCTGG GGACCCTCCT
121 TCCCTCGATC TCCTCGTGCG GGGTCTCCGA CGCGCAGACG CGGTGCGTGC GAGCGTGCAC
181 GTGCTGTCGC CGTGGCTGTA GTTGCGAGCG GAGAGAGAGA GAGAGGGAGG GAGGGAGGGC
241 AGAGGGCAGA GGACATGCGG GTGGGAACGT GCACCGGCCT CGTCTCACGC AGCTGGAGCC
301 CACGAATGCC ACCACCACAC CCTTCTTCCC CCCCCCTCCC TCTTCTTCCC ACGGCGGCGA
361 CGACGACGGG GGCTCAGCTC ACGCTTCTGT AGGGTATTAT ATTAAAGCAC ATGTGCGTGC
421 TGTGCTTCGG CTTCTTGTTG GTGGTCGTTC GCTTTCAAGT CCACGACTCC TGCCGTTGCT
481 CACCGCCGGG CTGTCTTCTT TCGCCCCTCC CTCGGTGCCT CTTCCGCCGC CGCACGCGTG
541 CGCTGATCAC GCGTCTTGCG TGATGTGCCT GTGTGTATAC ACACCGTGCA CGGAGAGAGC
601 GAGCGAGCGA GAGAGAGAGA GGGCGAGAGG AAGAATGGCC GGTGCCTCGC GTGGGCAAGC
661 GTGCGCGAGT TTGTCGTGTC GCTGCGCCGC GTGGATGTCG CGCAGCGCAA GACCCCGTGC
721 GCCTGAAACG TGAAGGGAGG TCGAGAAGCG TGCCGCATGA GGCCTTAAGG CAGGAAAGTG
781 AAAAGAGCTC GACGCGAGTG CAGCACGCGC GACATCGAAC AGGACGGAAT AGACACTCAG
841 CCCCTTCCCC TTTCAAAAAC TGAATGGACG GACATCGTAA CGCGCTTCTC TCCCCTCCAC
901 TCCCCCTGTC CCTGTCTCCT TGACGTGTGC TGCTCTCGTG AGCGACAGAT GAAGCGAGGC
961 GTAGAAGGAA AGGGGGGGGA GCGAGGCCTG CACACACACA CACACACACA CACACACACA
1021 CACACACACA CACACACACA CACACACGAA AAGCAAGTTT GAGGCAGACG TGGTTTCACA
1081 CGATTCCTGT GCAAGAAGCG TGCCTGCGTG ACGGTGTGGC TGTGTATGCA TGCGTGAAGG
1141 GTTACCGCAA CGATGCGCAA TGGACTCCCC CGCTTTCCAT TTCGTCACCT TCCGCCTCTC
1201 TCTCTCTCTC TCTCACCATC TACGCGTGCA CATCATCAAC TGTCTCTTGT CGGTGCTCAC
1261 CACCCTCAAC CACCCCTCAC TTTCAAGGCT TCCCGAACGC ACACAAAAGG CGTGAAAACC
1321 GCTCGCGTGT GTTGAGCCGT CCACCGTAGC CCTCCCCCTG TCCCCGGGGG ATCCGGCCGT
1381 GCCGGCCGAG ATATCGTGAG CAAGGGCGAG GAGCTGTTCA CCGGGGTGGT GCCCATCCTG
1441 GTCGAGCTGG ACGGCGACGT GAACGGCCAC AAGTTCAGCG TGTCCGGCGA GGGCGAGGGC
1501 GATGCCACCT ACGGCAAGCT GACCCTGAAG TTCATCTGCA CCACCGGCAA GCTGCCCGTG
1561 CCCTGGCCCA CCCTCGTGAC CACCTTCACC TACGGCGTGC AGTGCTTCAG CCGCTACCCC
1621 GACCACATGA AGCAGCACGA CTTCTTCAAG TCCGCCATGC CCGAAGGCTA CGTCCAGGAG
1681 CGCACCATCT TCTTCAAGGA CGACGGCAAC TACAAGACCC GCGCCGAGGT GAAGTTCGAG
1741 GGCGACACCC TGGTGAACCG CATCGAGCTG AAGGGCATCG ACTTCAAGGA GGACGGCAAC
1801 ATCCTGGGGC ACAAGCTGGA GTACAACTAC AACAGCCACA ACGTCTATAT CATGGCCGAC
1861 AAGCAGAAGA ACGGCATCAA GGTGAACTTC AAGATCCGCC ACAACATCGA GGACGGCAGC
1921 GTGCAGCTCG CCGACCACTA CCAGCAGAAC ACCCCCATCG GCGACGGCCC CGTGCTGCTG
1981 CCCGACAACC ACTACCTGAG CACCCAGTCC GCCCTGAGCA AAGACCCCAA CGAGAAGCGC
2041 GATCACATGG TCCTGCTGGA GTTCGTCTCG CTGACCGCCG CCGGGATCAC TCACGGCATG
2101 GACGAGCTGT ACAAGTAAAG ATCCACTAGT TCTAGTTCTA GGGGGCGAGC TCGAATTCAG
2161 ATCCTCGTGT GAGCGTTCGC GGAATCGGTC GCTCGTGTTT ATGCCCGTCT TGGTGTTGTG
2221 CTCGCAAGGC GGTGCAGCAG GATACCGTCG CCCTCCTCTC TCCTTGCTTC TCTGTTCTTC
2281 AATTCGCGAT CTCACAGAGG CCGGCTGTGC ACGCCCTTCC TCACCCTCCT TTTCCCACCT
2341 CTCGGCCACC GGTCGGCTCC GTTCCGTCTG CCGTCGAGAA GGGACGGGCA TGTGCAGCTC
2401 CTCCCTTTCT CTCGCGCGCG CATCTTCTCT TGCTTGTGGC ACTCACGCTC ATGCGTCAAG
2461 GCGGCCCCAC GCGAGCCCCT GCGCTCCCTT CCCTCTTGCG CATCCGTAGC CGGACTGGTC
2521 GATGCGCAAG GCCGGCATGA AGGAGCGCGT GCCCTCAAGA GCGGACTATC ATGCCCTACG
2581 TGGGCCACGC AGCGATGAGG CCGGCTTCGC GGAGATGCGT CACGCACGTG CCAGATGATG
2641 CCGTACGCCT TCCTTGACTT GCGCCCCCCT CTCTTCCTCC GTCTCTCACT CTCTCTCTCT
2701 CACACACACA CACACACACA CACACACACA AAGCTTCCTT GCACACAACG TCCCCCTCGT
2761 TGTGCGGGCT GACGCCTCCC GTTGACAACT TGCCTTACGC ACGCAGCACT CGGCCAGCCA
2821 ACGAGGCCGC AGCGGCATCA CGTGTATCTT TCCACAGAAA CCGCGTGCTT TCGGATACTC
2881 TAGTATTTTT GTTTGTGGTC GAATCTCCCT TTAGTGAGGG TTAATTAGTC CTGCATTAAT
2941 GAATCGGCCA ACGCGCGGGG AGAGGCGGTT TGCGTATTGG GCGCTCTTCC GCTTCCTCGC
3001 TCACTGACTC GCTGCGCTCG GTCGTTCGGC TGCGGCGAGC GGTATCAGCT CACTCAAAGG
3061 CGGTAATACG GTTATCCACA GAATCAGGGG ATAACGCAGG AAAGAACATG TGAGCAAAAG
3121 GCCAGCAAAA GGCCAGGAAC CGTAAAAAGG CCGCGTTGCT GGCGTTTTTC CATAGGCTCC
3181 GCCCCCCTGA CGAGCATCAC AAAAATCGAC GCTCAAGTCA GAGGTGGCGA AACCCGACAG
3241 GACTATAAAG ATACCAGGCG TTTCCCCCTG GAAGCTCCCT CGTGCGCTCT CCTGTTCCGA
3301 CCCTGCCGCT TACCGGATAC CTGTCCGCCT TTCTCCCTTC GGGAAGCGTG GCGCTTTCTC
3361 AATGCTCACG CTGTAGGTAT CTCAGTTCGG TGTAGGTCGT TCGCTCCAAG CTGGGCTGTG
3421 TGCACGAACC CCCCGTTCAG CCCGACCGCT GCGCCTTATC CGGTAACTAT CGTCTTGAGT
3481 CCAACCCGGT AAGACACGAC TTATCGCCAC TGGCAGCAGC CACTGGTAAC AGGATTAGCA
3541 GAGCGAGGTA TGTAGGCGGT GCTACAGAGT TCTTGAAGTG GTGGCCTAAC TACGGCTACA
3601 CTAGAAGGAC AGTATTTGGT ATCTGCGCTC TGCTGAAGCC AGTTACCTTC GGAAAAAGAG
3661 TTGGTAGCTC TTGATCCGGC AAACAAACCA CCGCTGGTAG CGGTGGTTTT TTTGTTTGCA
3721 AGCAGCAGAT TACGCGCAGA AAAAAAGGAT CTCAAGAAGA TCCTTTGATC TTTTCTACGG
3781 GGTCTGACGC TCAGTGGAAC GAAAACTCAC GTTAAGGGAT TTTGGTCATG AGATTATCAA
3841 AAAGGATCTT CACCTAGATC CTTTTAAATT AAAAATGAAG TTTTAAATCA ATCTAAAGTA
3901 TATATGAGTA AACTTGGTCT GACAGTTACC AATGCTTAAT CAGTGAGGCA CCTATCTCAG
3961 CGATCTGTCT ATTTCGTTCA TCCATAGTTG CCTGACTCCC CGTCGTGTAG ATAACTACGA
4021 TACGGGAGGG CTTACCATCT GGCCCCAGTG CTGCAATGAT ACCGCGAGAC CCACGCTCAC
4081 CGGCTCCAGA TTTATCAGCA ATAAACCAGC CAGCCGGAAG GGCCGAGCGC AGAAGTGGTC
4141 CTGCAACTTT ATCCGCCTCC ATCCAGTCTA TTAATTGTTG CCGGGAAGCT AGAGTAAGTA
4201 GTTCGCCAGT TAATAGTTTG CGCAACGTTG TTGCCATTGC TACAGGCATC GTGGTGTCAC
4261 GCTCGTCGTT TGGTATGGCT TCATTCAGCT CCGGTTCCCA ACGATCAAGG CGAGTTACAT
4321 GATCCCCCAT GTTGTGCAAA AAAGCGGTTA GCTCCTTCGG TCCTCCGATC GTTGTCAGAA
4381 GTAAGTTGGC CGCAGTGTTA TCACTCATGG TTATGGCAGC ACTGCATAAT TCTCTTACTG
4441 TCATGCCATC CGTAAGATGC TTTTCTGTGA CTGGTGAGTA CTCAACCAAG TCATTCTGAG
4501 AATAGTGTAT GCGGCGACCG AGTTGCTCTT GCCCGGCGTC AATACGGGAT AATACCGCGC
4561 CACATAGCAG AACTTTAAAA GTGCTCATCA TTGGAAAACG TTCTTCGGGG CGAAAACTCT
4621 CAAGGATCTT ACCGCTGTTG AGATCCAGTT CGATGTAACC CACTCGTGCA CCCAACTGAT
4681 CTTCAGCATC TTTTACTTTC ACCAGCGTTT CTGGGTGAGC AAAAACAGGA AGGCAAAATG
4741 CCGCAAAAAA GGGAATAAGG GCGACACGGA AATGTTGAAT ACTCATACTC TTCCTTTTTC
4801 AATATTATTG AAGCATTTAT CAGGGTTATT GTCTCATGAG CGGATACATA TTTGAATGTA
4861 TTTAGAAAAA TAAACAAATA GGGGTTCCGC GCACATTTCC CCGAAAAGTG CCACCTGACG
4921 TCTAAGAAAC CATTATTATC ATGACATTAA CCTATAAAAA TAGGCGTATC ACGAGGCCCT
4981 TTCGTCTCGC GCGTTTCGGT GATGACGGTG AAAACCTCTG ACACATGCAG CTCCCGGAGA
5041 CGGTCACAGC TTGTCTGTAA GCGGATGCCG GGAGCAGACA AGCCCGTCAG GGCGCGTCAG
5101 CGGGTGTTGG CGGGTGTCGG GGCTGGCTTA ACTATGCGGC ATCAGAGCAG ATTGTACTGA
5161 GAGTGCACCA TATCGACGCT CTCCCTTATG CGACTCCTGC ATTAGGAAGC AGCCCAGTAG
5221 TAGGTTGAGG CCGTTGAGCA CCGCCGCCGC AAGGAATGGT GCATGCAAGG AGATGGCGCC
5281 CAACAGTCCC CCGGCCACGG GGCCTGCCAC CATACCCACG CCGAAACAAG CGCTCATGAG
5341 CCCGAAGTGG CGAGCCCGAT CTTCCCCATC GGTGATGTCG GCGATATAGG CGCCAGCAAC
5401 CGCACCTGTG GCGCCGGTGA TGCCGGCCAC GATGCGTCCG GCGTAGAGGA TCTGGCTAGC
5461 GATGACCCTG CTGATTGGTT CGCTGACCAT TTCCGGGGTG CGGAACGGCG TTACCAGAAA
5521 CTCAGAAGGT TCGTCCAACC AAACCGACTC TGACGGCAGT TTACGAGAGA GATGATAGGG
5581 TCTGCTTCAG TAAGCCAGAT GCTACACAAT TAGGCTTGTA CATATTGTCG TTAGAACGCG
5641 GCTACAATTA ATACATAACC TTATGTATCA TACACATACG ATTTAGGTGA CACTATAGAA
5701 TACAAGCTTG GGCTGCAGGT CGACGGGGTG ATGGAGAGAA TGGTTGACCA TGAGGACGCC
5761 GATGGGCGAT GCGCCAGCGC GTGCTTGCGT ACTTCAATCG TTTTTCATCT TGATTCAGAG
5821 CTAAGTAGTA GCACACCACA GCAAGCGTAA TCATGTAGCA TGTATGCAGG TGCACGAGTA
5881 GGCACTCTCT GTCTCTGTCC GGCATCTATG ATGCTCTCCG GCCGCGTGCG CGCGACGCCT
5941 CTGTGTGAGT GCTCGTGTGA GTGTGTGAGA GCTGTCCAGG CCTTTCCTCT GCCTACTCGT
6001 CCCACGCTCA GGCAGGCACA GAGGAATGAA AAGGAAGACG AGGACGCAAC GCCAGGCGCA
6061 TCGACATCGC GCACGTGTTT GCGGTGTACT GTGCGTCGAT GATCGAGCGG CCAAGCGTCC
6121 AGCTTCACGC GAGGAGGACT GCATCGCTGC TCCTGTGCGC GTGCGTGCAC GCGTCAAGGC
6181 ACGCTCGATT CGTTCCCAGG GTGGATACCC TCCCCCCTCC CCCAGCTGCA GACAACCCCC
6241 ACCCTCTCGA GGCTTGCACC CAGGCTCGTC AGTCTCTCTA CCACTTTCTG CCTTCTGTAG
6301 TGCACCCCAC CCCTGCATTC TCCTCTCTCT CACACACACA CACACACACA CACACACACA
6361 CACACACACA CGTACACACG TACACACACA CGTGCGCATG CTTGACGCAT ACGCGACGAA
6421 TTCGAAAGCT CACCTCATTC CTCCCTCCTC ACACCATCAT CGGCATCCAT AGAGACACGT
6481 GCGCGTAGAA CGATACAGCT CACGCGCACA GAGAGGCGTG TTCTGGTCGT GCGCATCATC
6541 GGCACAGCAC TGGCGGACGA AACTCGCACA CAGGCACGCC GCCTCCTTTC ACCCGTCATA
6601 GATAGTTGAA TTAGACGCCC TCCTCCTCCC TCATCATCGC CGTCGTCATC CGGGTCCGAG
6661 CACTAGTCGC GTGTGGCCTC GAACACCGAG CGACCCTGCA GCCAATATGG GATCGGCCAT
6721 TGAACAAGAT GGATTGCACG CAGGTTCTCC GGCCGCTTGG GTGGAGAGGC TATTCGGCTA
6781 TGACTGGGCA CAACAGACAA TCGGCTGCTC TGATGCCGCC GTGTTCCGGC TGTCAGCGCA
6841 GGGGCGCCCG GTTCTTTTTG TCAAGACCGA CCTGTCCGGT GCCCTGAATG AACTGCAGGA
6901 CGAGGCAGCG CGGCTATCGT GGCTGGCCAC GACGGGCGTT CCTTGCGCAG CTGTGCTCGA
6961 CGTTGTCACT GAAGCGGGAA GGGACTGGCT GCTATTGGGC GAAGTGCCGG GGCAGGATCT
7021 CCTGTCATCT CACCTTGCTC CTGCCGAGAA AGTATCCATC ATGGCTGATG CAATGCGGCG
7081 GCTGCATACG CTTGATCCGG CTACCTGCCC ATTCGACCAC CAAGCGAAAC ATCGCATCGA
7141 GCGAGCACGT ACTCGGATGG AAGCCGGTCT TGTCGATCAG GATGATCTGG ACGAAGAGCA
7201 TCAGGGGCTC GCGCCAGCCG AACTGTTCGC CAGGCTCAAG GCGCGCATGC CCGACGGCGA
7261 GGATCTCGTC GTGACCCATG GCGATGCCTG CTTGCCGAAT ATCATGGTGG AAAATGGCCG
7321 CTTTTCTGGA TTCATCGACT GTGGCCGGCT GGGTGTGGCG GACCGCTATC AGGACATAGC
7381 GTTGGCTACC CGTGATATTG CTGAAGAGCT TGGCGGCGAA TGGGCTGACC GCTTCCTCGT
7441 GCTTTACGGT ATCGCCGCTC CCGATTCGCA GCGCATCGCC TTCTATCGCC TTCTTGACGA
7501 GTTCTTCTGA GGGGATCGGC AATAAAAAGA CAGAATAAAA CGCACGGGTG TTGGGTCGTT
7561 TGTTCGGATC
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B2863 pXG-/GFP+