Target-serine-threonine+phosophatase+2C+Toxoplasma+Gondii

**ORGANISM : ** Toxoplasma gondii ME49

**GENE of INTEREST: ** TGME49_031850 (serine-threonine phosophatase 2C)

**BACKGROUND/ DISEASE: **

**IMAGE: **

**NCBI GENE # or RefSeq#**: 7898899

**EC# OF ENZYME: **  3 .1.3.16

**CDS GENE SEQUENCE: ** code format="genbank" 1 atgaagtcct ctgctgaaat taggcggacc atggatgtcc ctcctaccat tcatgtacct 61 ctccctccaa cgtcgtatcc cgctttcgat gctgcgatct tcacagacat cggtgggcgc 121 aagcatcagg aagataggtt cactctctgt ccgcagctcg ttcccggccg agacgactgc 181 gccttcttcg gtgtcttcga tggcactgtt ggagatttcg ccagcgaaaa tgtgaaggat 241 cttgttgttc cacagttgat ttcctcgccc gcgtggcagg aggtgactga gatgctgaga 301 tcagacgtac ccgccactga ggtggacgag aagctccctc agttgcttga tcaggcagtc 361 gatgacatgt acaagaacgc agacaacgaa cttgtgaaga tgtgcgagca gcttaacaaa 421 gactacgcga gcagcacctc cgtcacggca gtcttggcca aaggcttcgt ggctgttggt 481 catctgggcg acagccgcat cgctatggga gtcgagacgc caaacgggtt gaactgcgag 541 ttcttgaccg ttgaccacaa gccggatatg ccacatgaga aactgcgcat catgcgcaat 601 ggaggcagtg ttgagtatct ccacaaccac aacaacaaac cgttcattcg aggtggtgac 661 ttctcgttcc ggaagtcgcg cggagagcag ccgatgcagc tccagtactc ccgagctttc 721 ggcgggaagg acctgaagat gtacggtttg agcaaccaac ccgacgtacg cgttgtccgc 781 gtcacgcctc aacaccgagt gatgattctc gcgactgatg gcttgtggga cgtcatgtct 841 gcggcgcaag ctgtagagat cgctatgcag gcacgacagg aaggaaggaa cccagcgcag 901 gcgctggtgg agatgaccct cgctgagcag cagagccgca accaaagtgc agacaacatt 961 actgcgatga cagtgttctt caagaagact gattag code

**AMINO ACID SEQUENCE: **

SLTMDVPPTIHVPLPPTSYPAFDAAIFTDIGGRKHQEDRFTLCPQLVPGRDDCAFFGVFDGTVGDFASENVKD

LVVPQLISSPAWQEVTEMLRSDVPATEVDEKLPQLLDQAVDDMYKNADNELVKMCEQLNKDYASSTSVTAVLA

KGFVAVGHLGDSRIAMGVETPNGLNCEFLTVDHKPDMPHEKLRIMRNGGSVEYLHNHNNKPFIRGGDFSFRKS

RGEQPMQLQYSRAFGGKDLKMYGLSNQPDVRVVRVTPQHRVMILATDGLWDVMSAAQAVEIAMQARQEGRNPA

QALVEMTLAEQQSRNQSADNTAMTVFFKKTD

[]
 * PDB:**

Primer Design - Larry

Upstream 5’ __TACTTCCAATCCATG__AAGTCCTCTGCTGAAAT 3’

Downstream 5’ __TATCCACCTTTACTGCTA__ATCAGTCTTCTTGAAGA 3’ Downstream Reverse Complement 5’ TCTTCAAGAAGACTGAT__TAGCAGTAAAGGTGGATA__ 3’

1 ATGAAGTCCT CTGCTGAAAT TAGGCGGACC ATGGATGTCC CTCCTACCAT TCATGTACCT 61 CTCCCTCCAA CGTCGTATCC CGCTTTCGAT GCTGCGATCT TCACAGACAT CGGTGGGCGC 121 AAGCATCAGG AAGATAGGTT CACTCTCTGT CCGCAGCTCG TTCCCGGCCG AGACGACTGC 181 GCCTTCTTCG GTGTCTTCGA TGGCACTGTT GGAGATTTCG CCAGCGAAAA TGTGAAGGAT 241 CTTGTTGTTC CACAGTTGAT TTCCTCGCCC GCGTGGCAGG AGGTGACTGA GATGCTGAGA 301 TCAGACGTAC CCGCCACTGA GGTGGACGAG AAGCTCCCTC AGTTGCTTGA TCAGGCAGTC 361 GATGACATGT ACAAGAACGC AGACAACGAA CTTGTGAAGA TGTGCGAGCA GCTTAACAAA 421 GACTACGCGA GCAGCACCTC CGTCACGGCA GTCTTGGCCA AAGGCTTCGT GGCTGTTGGT 481 CATCTGGGCG ACAGCCGCAT CGCTATGGGA GTCGAGACGC CAAACGGGTT GAACTGCGAG 541 TTCTTGACCG TTGACCACAA GCCGGATATG CCACATGAGA AACTGCGCAT CATGCGCAAT 601 GGAGGCAGTG TTGAGTATCT CCACAACCAC AACAACAAAC CGTTCATTCG AGGTGGTGAC 661 TTCTCGTTCC GGAAGTCGCG CGGAGAGCAG CCGATGCAGC TCCAGTACTC CCGAGCTTTC 721 GGCGGGAAGG ACCTGAAGAT GTACGGTTTG AGCAACCAAC CCGACGTACG CGTTGTCCGC 781 GTCACGCCTC AACACCGAGT GATGATTCTC GCGACTGATG GCTTGTGGGA CGTCATGTCT 841 GCGGCGCAAG CTGTAGAGAT CGCTATGCAG GCACGACAGG AAGGAAGGAA CCCAGCGCAG 901 GCGCTGGTGG AGATGACCCT CGCTGAGCAG CAGAGCCGCA ACCAAAGTGC AGACAACATT 961 ACTGCGATGA CAGTGTTCTT CAAGAAGACT GATTAG

Melting Temp
 * || GC Content || 0mM Mg++ || 1.5mM Mg++ || 2mM Mg++ || 4mM Mg++ || 6mM Mg++ ||
 * Forward || 40.0 % || 52.0 ºC || 59.0 ºC || 59.8 ºC || 61.3 ºC || 62.0 ºC ||
 * Reverse || 35.0 % || 47.0 ºC || 54.2 ºC || 55.0 ºC || 56.5 ºC || 57.2 ºC ||

**Primer Design - Justin**

<span style="font-family: Arial,Helvetica,sans-serif; font-size: 12px;">**__Forward Primer:__** <span style="font-family: Arial,Helvetica,sans-serif; font-size: 12px;"> 5’ __ TACTTCCAATCCATG ____GATGTCCCGCCGACCATC__ 3’ __33__ bp <span style="display: block; font-family: Arial,Helvetica,sans-serif; font-size: 12px; text-align: center;">GC Content __54.5__% <span style="font-family: Arial,Helvetica,sans-serif; font-size: 12px;">0 mM Mg2+ Tm __67.0__ oC 1.5 mM Mg2+ Tm __73.5__ oC 2 mM Mg2+ Tm __73.9__ oC <span style="font-family: Arial,Helvetica,sans-serif; font-size: 12px;">4 mM Mg2+ Tm __74.8__ oC 6 mM Mg2+ Tm __75.1__ oC

<span style="font-family: Arial,Helvetica,sans-serif; font-size: 12px;">**__Reverse Primer__:** <span style="font-family: Arial,Helvetica,sans-serif; font-size: 12px;"> 5’ ____ 3’

<span style="font-family: Arial,Helvetica,sans-serif; font-size: 12px;">**__Reverse Complemented:__** <span style="font-family: Arial,Helvetica,sans-serif; font-size: 12px;"><span style="font-family: Arial,Helvetica,sans-serif; font-size: 12px;"> 5’ __ TATCCACCTTTACTG __ 3’ __33__ b GC Content __33.3__% <span style="font-family: Arial,Helvetica,sans-serif; font-size: 12px;">0 mM Mg2+ Tm __58.7__ oC 1.5 mM Mg2+ Tm __66.8__ oC 2 mM Mg2+ Tm __67.3__ oC <span style="font-family: Arial,Helvetica,sans-serif; font-size: 12px;">4 mM Mg2+ Tm __68.5__ oC 6 mM Mg2+ Tm __69.0__ oC

<span style="font-family: Arial,Helvetica,sans-serif; font-size: 12px;">Primer Design - Jericho


 * OUTPUT FROM DNA WORKS**

TCTCTGACCATGGATGTACCACCTACCATTCACGTTCCTCTGCCGCCGACGTCTTACCCG GCGTTCGACGCGGCGATCTTCACCGACATCGGTGGCCGTAAACACCAGGAAGACCGCTTC ACCCTGTGCCCGCAGCTGGTTCCGGGTCGTGACGACTGCGCGTTCTTCGGTGTTTTCGAT GGTACCGTTGGTGACTTCGCGTCTGAAAACGTTAAGGACCTGGTTGTTCCTCAGCTCATC TCTTCTCCGGCGTGGCAGGAAGTGACCGAAATGCTGCGTTCTGATGTTCCAGCCACTGAG GTTGATGAGAAACTGCCGCAACTGCTGGACCAGGCGGTAGACGACATGTACAAAAACGCG GACAACGAGCTGGTGAAAATGTGCGAACAGCTGAACAAAGACTACGCGTCTTCTACCTCT GTTACCGCCGTTCTGGCGAAAGGTTTCGTTGCGGTTGGTCACCTGGGTGACTCTCGTATT GCGATGGGTGTTGAAACCCCGAACGGTCTCAACTGCGAATTCCTGACCGTTGACCACAAA CCGGACATGCCGCACGAAAAACTGCGTATTATGCGTAACGGTGGTTCTGTTGAATACCTG CACAACCACAACAACAAGCCGTTCATCCGTGGTGGCGACTTCTCTTTCCGTAAATCTCGT GGTGAACAGCCGATGCAGCTCCAGTATTCTCGTGCGTTCGGTGGTAAAGACCTCAAAATG TACGGTCTGTCTAACCAGCCGGACGTTCGTGTTGTGCGTGTTACTCCGCAGCACCGTGTT ATGATCCTGGCGACCGATGGTCTGTGGGACGTTATGTCTGCGGCGCAGGCCGTTGAGATT GCCATGCAAGCACGTCAAGAAGGTCGTAATCCGGCGCAAGCGCTCGTCGAAATGACCCTG GCGGAACAGCAGTCTCGTAACCAGAGCGCCGACAACACCGCGATGACGGTTTTCTTCAAA AAAACCGACTAA

Melting Temperatures

Upstream: 5’ TACTTCCAATCCATGATGGATGTACCACCTACCATT 3’ __36__ bp GC Content __41.7__ %

0 mM Mg2+ Tm __63.0__ ºC 1.5 mM Mg2+ Tm __70.5__ ºC 2 mM Mg2+ Tm __71.0__ ºC 4 mM Mg2+ Tm __72.0__ ºC 6 mM Mg2+ Tm __72.4__ ºC

Downstream: 5’ TTCTTCAAAAAAACCGACTAACAGTAAAGGTGGATA 3’

Reverse Complement: 5’ TATCCACCTTTACTGTTAGTCGGTTTTTTTGAAGAA 3’ __36__ bp GC Content __33.3__ %

0 mM Mg2+ Tm __59.9__ ºC 1.5 mM Mg2+ Tm __68.1__ ºC 2 mM Mg2+ Tm __68.6__ ºC 4 mM Mg2+ Tm __69.7__ ºC 6 mM Mg2+ Tm __70.2__ ºC