Molecular Tech Fall 2017 Human Malate Dehydrogenase 2 Info
MDH2Human [>sp|P40926|25-338]. Without Transit SequenceC term His tag plus TEV
MAKVAVLGAS GGIGQPLSLL LKNSPLVSRL TLYDIAHTPG VAADLSHIET
KAAVKGYLGP EQLPDCLKGC DVVVIPAGVP RKPGMTRDDL FNTNATIVAT
LTAACAQHCP EAMICVIANP VNSTIPITAE VFKKHGVYNP NKIFGVTTLD
IVRANTFVAE LKGLDPARVN VPVIGGHAGK TIIPLISQCT PKVDFPQDQL
TALTGRIQEA GTEVVKAKAG AGSATLSMAY AGARFVFSLV DAMNGKEGVV
ECSFVKSQET ECTYFSTPLL LGKKGIEKNL GIGKVSSFEE KMISDAIPEL
KASIKKGEDF VKTLKENLYF QGHHHHHH
TEV cleavage site is shown in red
6xHis-tag is shown in blue
cDNA Sequence of MDH2 (Codon-optimized for expression in BL21 (DE3).
CCATGGCTAAAGTAGCTGTTCTGGGTGCATCTGGTGGTATTGGTCAACCGCTGTCCCTGCTGCTGAAGAACTCTCCACTGGTTAGCCGTCTGACCCTGTACGATATCGCACATACTCCAGGCGTTGCTGCTGACCTGTCCCACATTGAGACTAAAGCTGCGGTGAAAGGCTACCTGGGTCCAGAACAGCTGCCAGATTGTCTGAAAGGTTGTGACGTGGTAGTTATCCCAGCAGGTGTTCCACGTAAACCAGGTATGACCCGTGATGACCTGTTCAATACTAATGCGACTATCGTTGCGACCCTGACCGCTGCTTGCGCTCAGCACTGTCCGGAAGCTATGATCTGTGTGATTGCTAATCCGGTTAACAGCACCATCCCGATTACTGCCGAGGTATTCAAGAAACACGGTGTTTACAACCCGAACAAGATCTTCGGTGTTACCACCCTGGATATCGTGCGTGCAAACACCTTTGTGGCTGAACTGAAAGGTCTGGATCCAGCTCGCGTAAACGTTCCGGTAATCGGTGGTCACGCTGGCAAGACCATCATTCCGCTGATTTCCCAGTGCACTCCGAAAGTAGACTTCCCACAGGACCAGCTGACCGCTCTGACTGGTCGTATCCAGGAGGCAGGCACTGAAGTGGTGAAAGCGAAAGCAGGTGCTGGCTCTGCGACTCTGAGCATGGCCTACGCTGGTGCTCGTTTCGTATTCTCTCTGGTTGACGCAATGAACGGCAAGGAAGGTGTAGTGGAATGTTCCTTCGTGAAGTCTCAAGAAACCGAATGCACCTACTTCAGCACTCCACTGCTGCTGGGCAAGAAAGGCATCGAGAAGAACCTGGGCATCGGTAAAGTTTCTTCTTTCGAAGAGAAGATGATCTCCGACGCAATTCCGGAACTGAAAGCGAGCATCAAGAAGGGTGAAGATTTCGTGAAGACCCTGAAAGAGAACCTGTACTTCCAGGGTCATCATCACCACCATCACTAACTCGAG
Map of hMDH2/AA25-338
Synthetic gene was cloned into NcoI/XhoIdigested pET28a.
Open reading frame orientation as illustrated. Not all unique restriction sites are shown in the map. Extra nucleotides or unique restriction sites may be found on both ends of your gene for subcloning purpose.
Seq:
LOCUS GS61493-1 pET28a-hMDH2_AA25-338 6219 bpds-DNA circular SYN 25-Jun-2017
DEFINITION .
ACCESSION .
VERSION .
KEYWORDS GS61493-1 pET28a-hMDH2_AA25-338
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6219)
AUTHORS .
TITLE DirectSubmission
FEATURES Location/Qualifiers
source 1..6219
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 26..73
/note="T7 terminator"
/note="transcriptionterminatorforbacteriophage T7 RNA
polymerase"
CDS complement(140..157)
/codon_start=1
/product="6xHis affinitytag"
/note="6xHis"
/translation="HHHHHH"
gene complement(158..1152)
/note="hMDH2_AA25-338"
CDS complement(167..184)
/codon_start=1
/product="6xHis affinitytag"
/note="6xHis"
/translation="HHHHHH"
protein_bind 1194..1218
/bound_moiety="lacrepressorencodedbylacI"
/note="lacoperator"
/note="Thelacrepressorbinds to thelacoperator to
inhibittranscription in E. coli. Thisinhibition can be
relievedbyaddinglactoseor
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(1219..1237)
/note="T7 promoter"
/note="promoterforbacteriophage T7 RNA polymerase"
promoter 1546..1623
/gene="lacI"
/note="lacIpromoter"
/note="
"
CDS 1624..2706
/codon_start=1
/gene="lacI"
/product="lacrepressor"
/note="lacI"
/note="Thelacrepressorbinds to thelacoperator to
inhibittranscription in E. coli. Thisinhibition can be
relievedbyaddinglactoseor
ORIGIN
1 atccggatatagttcctcctttcagcaaaaaacccctcaagacccgtttagaggccccaa
61 ggggttatgctagttattgctcagcggtggcagcagccaactcagcttcctttcgggctt
121 tgttagcagccggatctcagtggtggtggtggtggtgctcgagttagtgatggtggtgat
181 gatgaccctggaagtacaggttctctttcagggtcttcacgaaatcttcacccttcttga
241 tgctcgctttcagttccggaattgcgtcggagatcatcttctcttcgaaagaagaaactt
301 taccgatgcccaggttcttctcgatgcctttcttgcccagcagcagtggagtgctgaagt
361 aggtgcattcggtttcttgagacttcacgaaggaacattccactacaccttccttgccgt
421 tcattgcgtcaaccagagagaatacgaaacgagcaccagcgtaggccatgctcagagtcg
481 cagagccagcacctgctttcgctttcaccacttcagtgcctgcctcctggatacgaccag
541 tcagagcggtcagctggtcctgtgggaagtctactttcggagtgcactgggaaatcagcg
601 gaatgatggtcttgccagcgtgaccaccgattaccggaacgtttacgcgagctggatcca
661 gacctttcagttcagccacaaaggtgtttgcacgcacgatatccagggtggtaacaccga
721 agatcttgttcgggttgtaaacaccgtgtttcttgaatacctcggcagtaatcgggatgg
781 tgctgttaaccggattagcaatcacacagatcatagcttccggacagtgctgagcgcaag
841 cagcggtcagggtcgcaacgatagtcgcattagtattgaacaggtcatcacgggtcatac
901 ctggtttacgtggaacacctgctgggataactaccacgtcacaacctttcagacaatctg
961 gcagctgttctggacccaggtagcctttcaccgcagctttagtctcaatgtgggacaggt
1021 cagcagcaacgcctggagtatgtgcgatatcgtacagggtcagacggctaaccagtggag
1081 agttcttcagcagcagggacagcggttgaccaataccaccagatgcacccagaacagcta
1141 ctttagccatggtatatctccttcttaaagttaaacaaaattatttctagaggggaattg
1201 ttatccgctcacaattcccctatagtgagtcgtattaatttcgcgggatcgagatctcga
1261 tcctctacgccggacgcatcgtggccggcatcaccggcgccacaggtgcggttgctggcg
1321 cctatatcgccgacatcaccgatggggaagatcgggctcgccacttcgggctcatgagcg
1381 cttgtttcggcgtgggtatggtggcaggccccgtggccgggggactgttgggcgccatct
1441 ccttgcatgcaccattccttgcggcggcggtgctcaacggcctcaacctactactgggct
1501 gcttcctaatgcaggagtcgcataagggagagcgtcgagatcccggacaccatcgaatgg
1561 cgcaaaacctttcgcggtatggcatgatagcgcccggaagagagtcaattcagggtggtg
1621 aatgtgaaaccagtaacgttatacgatgtcgcagagtatgccggtgtctcttatcagacc
1681 gtttcccgcgtggtgaaccaggccagccacgtttctgcgaaaacgcgggaaaaagtggaa
1741 gcggcgatggcggagctgaattacattcccaaccgcgtggcacaacaactggcgggcaaa
1801 cagtcgttgctgattggcgttgccacctccagtctggccctgcacgcgccgtcgcaaatt
1861 gtcgcggcgattaaatctcgcgccgatcaactgggtgccagcgtggtggtgtcgatggta
1921 gaacgaagcggcgtcgaagcctgtaaagcggcggtgcacaatcttctcgcgcaacgcgtc
1981 agtgggctgatcattaactatccgctggatgaccaggatgccattgctgtggaagctgcc
2041 tgcactaatgttccggcgttatttcttgatgtctctgaccagacacccatcaacagtatt
2101 attttctcccatgaagacggtacgcgactgggcgtggagcatctggtcgcattgggtcac
2161 cagcaaatcgcgctgttagcgggcccattaagttctgtctcggcgcgtctgcgtctggct
2221 ggctggcataaatatctcactcgcaatcaaattcagccgatagcggaacgggaaggcgac
2281 tggagtgccatgtccggttttcaacaaaccatgcaaatgctgaatgagggcatcgttccc
2341 actgcgatgctggttgccaacgatcagatggcgctgggcgcaatgcgcgccattaccgag
2401 tccgggctgcgcgttggtgcggatatctcggtagtgggatacgacgataccgaagacagc
2461 tcatgttatatcccgccgttaaccaccatcaaacaggattttcgcctgctggggcaaacc
2521 agcgtggaccgcttgctgcaactctctcagggccaggcggtgaagggcaatcagctgttg
2581 cccgtctcactggtgaaaagaaaaaccaccctggcgcccaatacgcaaaccgcctctccc
2641 cgcgcgttggccgattcattaatgcagctggcacgacaggtttcccgactggaaagcggg
2701 cagtgagcgcaacgcaattaatgtaagttagctcactcattaggcaccgggatctcgacc
2761 gatgcccttgagagccttcaacccagtcagctccttccggtgggcgcggggcatgactat
2821 cgtcgccgcacttatgactgtcttctttatcatgcaactcgtaggacaggtgccggcagc
2881 gctctgggtcattttcggcgaggaccgctttcgctggagcgcgacgatgatcggcctgtc
2941 gcttgcggtattcggaatcttgcacgccctcgctcaagccttcgtcactggtcccgccac
3001 caaacgtttcggcgagaagcaggccattatcgccggcatggcggccccacgggtgcgcat
3061 gatcgtgctcctgtcgttgaggacccggctaggctggcggggttgccttactggttagca
3121 gaatgaatcaccgatacgcgagcgaacgtgaagcgactgctgctgcaaaacgtctgcgac
3181 ctgagcaacaacatgaatggtcttcggtttccgtgtttcgtaaagtctggaaacgcggaa
3241 gtcagcgccctgcaccattatgttccggatctgcatcgcaggatgctgctggctaccctg
3301 tggaacacctacatctgtattaacgaagcgctggcattgaccctgagtgatttttctctg
3361 gtcccgccgcatccataccgccagttgtttaccctcacaacgttccagtaaccgggcatg
3421 ttcatcatcagtaacccgtatcgtgagcatcctctctcgtttcatcggtatcattacccc
3481 catgaacagaaatcccccttacacggaggcatcagtgaccaaacaggaaaaaaccgccct
3541 taacatggcccgctttatcagaagccagacattaacgcttctggagaaactcaacgagct
3601 ggacgcggatgaacaggcagacatctgtgaatcgcttcacgaccacgctgatgagcttta
3661 ccgcagctgcctcgcgcgtttcggtgatgacggtgaaaacctctgacacatgcagctccc
3721 ggagacggtcacagcttgtctgtaagcggatgccgggagcagacaagcccgtcagggcgc
3781 gtcagcgggtgttggcgggtgtcggggcgcagccatgacccagtcacgtagcgatagcgg
3841 agtgtatactggcttaactatgcggcatcagagcagattgtactgagagtgcaccatata
3901 tgcggtgtgaaataccgcacagatgcgtaaggagaaaataccgcatcaggcgctcttccg
3961 cttcctcgctcactgactcgctgcgctcggtcgttcggctgcggcgagcggtatcagctc
4021 actcaaaggcggtaatacggttatccacagaatcaggggataacgcaggaaagaacatgt
4081 gagcaaaaggccagcaaaaggccaggaaccgtaaaaaggccgcgttgctggcgtttttcc
4141 ataggctccgcccccctgacgagcatcacaaaaatcgacgctcaagtcagaggtggcgaa
4201 acccgacaggactataaagataccaggcgtttccccctggaagctccctcgtgcgctctc
4261 ctgttccgaccctgccgcttaccggatacctgtccgcctttctcccttcgggaagcgtgg
4321 cgctttctcatagctcacgctgtaggtatctcagttcggtgtaggtcgttcgctccaagc
4381 tgggctgtgtgcacgaaccccccgttcagcccgaccgctgcgccttatccggtaactatc
4441 gtcttgagtccaacccggtaagacacgacttatcgccactggcagcagccactggtaaca
4501 ggattagcagagcgaggtatgtaggcggtgctacagagttcttgaagtggtggcctaact
4561 acggctacactagaaggacagtatttggtatctgcgctctgctgaagccagttaccttcg
4621 gaaaaagagttggtagctcttgatccggcaaacaaaccaccgctggtagcggtggttttt
4681 ttgtttgcaagcagcagattacgcgcagaaaaaaaggatctcaagaagatcctttgatct
4741 tttctacggggtctgacgctcagtggaacgaaaactcacgttaagggattttggtcatga
4801 acaataaaactgtctgcttacataaacagtaatacaaggggtgttatgagccatattcaa
4861 cgggaaacgtcttgctctaggccgcgattaaattccaacatggatgctgatttatatggg
4921 tataaatgggctcgcgataatgtcgggcaatcaggtgcgacaatctatcgattgtatggg
4981 aagcccgatgcgccagagttgtttctgaaacatggcaaaggtagcgttgccaatgatgtt
5041 acagatgagatggtcagactaaactggctgacggaatttatgcctcttccgaccatcaag
5101 cattttatccgtactcctgatgatgcatggttactcaccactgcgatccccgggaaaaca
5161 gcattccaggtattagaagaatatcctgattcaggtgaaaatattgttgatgcgctggca
5221 gtgttcctgcgccggttgcattcgattcctgtttgtaattgtccttttaacagcgatcgc
5281 gtatttcgtctcgctcaggcgcaatcacgaatgaataacggtttggttgatgcgagtgat
5341 tttgatgacgagcgtaatggctggcctgttgaacaagtctggaaagaaatgcataaactt
5401 ttgccattctcaccggattcagtcgtcactcatggtgatttctcacttgataaccttatt
5461 tttgacgaggggaaattaataggttgtattgatgttggacgagtcggaatcgcagaccga
5521 taccaggatcttgccatcctatggaactgcctcggtgagttttctccttcattacagaaa
5581 cggctttttcaaaaatatggtattgataatcctgatatgaataaattgcagtttcatttg
5641 atgctcgatgagtttttctaagaattaattcatgagcggatacatatttgaatgtattta
5701 gaaaaataaacaaataggggttccgcgcacatttccccgaaaagtgccacctaaattgta
5761 agcgttaatattttgttaaaattcgcgttaaatttttgttaaatcagctcattttttaac
5821 caataggccgaaatcggcaaaatcccttataaatcaaaagaatagaccgagatagggttg
5881 agtgttgttccagtttggaacaagagtccactattaaagaacgtggactccaacgtcaaa
5941 gggcgaaaaaccgtctatcagggcgatggcccactacgtgaaccatcaccctaatcaagt
6001 tttttggggtcgaggtgccgtaaagcactaaatcggaaccctaaagggagcccccgattt
6061 agagcttgacggggaaagccggcgaacgtggcgagaaaggaagggaagaaagcgaaagga
6121 gcgggcgctagggcgctggcaagtgtagcggtcacgctgcgcgtaaccaccacacccgcc
6181 gcgcttaatgcgccgctacagggcgcgtcccattcgcca
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