LOCUS       sequence-430166-        7164 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the glutathionine S-transferase with N-terminal His tag and 
            fused to the N-terminus of a mitogen-activated protein kinase 6 from
            Arabidopsis thaliana.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7164)
  TITLE     Shanbhag lab plasmids
REFERENCE   2  (bases 1 to 7164)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: His-GST-MPK6
FEATURES             Location/Qualifiers
     source          1..7164
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1851..1868
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1921..1939)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1935..1982
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2019..2474
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2106..2125)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2316..2337
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2566..3381)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2566..3381)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3289..3308
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3373..3392
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3503..4091
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3992..4011
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4245..4262
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4277..4416
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4342..4361)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4502..4524
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4518..4709)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5518..6600)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6562..6581
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6601..6678)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6884..6903
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6987..7006
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6987..7005
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7006..7030
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             7045..7067
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             7086..7103
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             7113..7130
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             join(7134..7164,1..623)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
ORIGIN
        1 aaggccttgt tcagccgacg cgtttacttt tggaatatct ggaagaaaag tatgaagaac
       61 atttatacga acgtgacgaa ggggacaagt ggcgtaataa aaaattcgaa ttagggcttg
      121 agtttccgaa tctgccttat tatattgatg gtgatgtgaa gttaacgcag tcaatggcca
      181 ttatacgcta tattgcggat aaacacaaca tgctgggtgg gtgcccgaaa gagcgcgctg
      241 aaatcagtat gcttgagggt gcggttctgg atattcgata cggcgtgtca cgcatcgcct
      301 atagtaaaga tttcgagacc ctgaaagtgg actttctctc aaaactgcct gaaatgttaa
      361 aaatgtttga ggatcggctg tgtcataaaa cctatttgaa tggggaccat gttacacacc
      421 ctgatttcat gctgtatgat gctcttgatg ttgtattata tatggacccc atgtgccttg
      481 acgccttccc taagctggtt tgtttcaaaa aacgtattga ggcaatacct caaattgata
      541 aatatttgaa aagcagtaaa tatattgctt ggccgctcca gggttggcaa gctacatttg
      601 gaggcggtga tcatccgcca aaagggggcg gtggtagcgg tggagggggt tctatggatg
      661 gtggctctgg acaacctgct gctgatacag aaatgaccga agccccgggt ggttttccgg
      721 cagccgctcc aagtccgcag atgcctggca tagaaaacat tcctgcgact ttatctcatg
      781 gaggccgttt tatccagtac aacatctttg gaaatatttt cgaggttacg gccaagtaca
      841 aaccgcccat catgcccatc ggtaaaggag catatggtat cgtttgctcc gcgatgaact
      901 ctgaaaccaa tgaatctgta gctattaaaa aaattgcaaa tgcgtttgac aataaaattg
      961 atgctaaacg cacgcttcgt gaaattaagc tgctccgtca tatggatcat gagaatatcg
     1021 tcgcaataag agatatcatc ccgcctcctc tgcgtaatgc ttttaatgat gtgtatattg
     1081 cctatgaact tatggataca gacttgcatc agatcatacg ctcgaatcaa gctctgtctg
     1141 aagagcattg ccagtacttc ctctatcaga tcttacgtgg tctcaagtac attcattctg
     1201 caaatgtact tcatcgtgat ctcaaaccca gtaatttatt actgaacgcg aactgtgatc
     1261 tgaaaatctg cgattttggc ttagcacgtg ttacttccga atcagacttc atgaccgagt
     1321 atgttgttac ccgttggtat cgtgcaccgg aacttctgtt gaatagctcg gactacactg
     1381 ctgcgattga tgtatggtct gttggctgta ttttcatgga attaatggat cgtaaacctc
     1441 tgtttccagg acgcgaccat gttcatcagt tacgactctt aatggaactg attggtactc
     1501 catcagagga agagttagaa tttcttaatg aaaatgccaa aagatacatt cgccagctgc
     1561 ctccgtatcc gcgtcagagt atcacagata agtttccaac ggttcatccg ctggccattg
     1621 accttatcga aaagatgtta acttttgacc cgcgtcggcg cataacagtg ctggatgccc
     1681 tggcccatcc ttatctgaat tcccttcacg atatttctga tgaaccggag tgcactatac
     1741 catttaattt tgatttcgaa aatcatgccc tgtcagagga acagatgaag gaacttattt
     1801 accgtgaagc gttggctttc aacccggagt accaacagtg ataactcgag caccaccacc
     1861 accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     1921 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     1981 tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt
     2041 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2101 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2161 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2221 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2281 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     2341 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     2401 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     2461 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2521 tttttctaaa tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc
     2581 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
     2641 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     2701 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     2761 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     2821 tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     2881 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     2941 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     3001 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     3061 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     3121 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     3181 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     3241 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     3301 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt
     3361 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
     3421 tattgttcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3481 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3541 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3601 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     3661 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3721 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3781 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3841 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3901 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3961 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4021 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4081 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4141 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     4201 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     4261 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     4321 accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     4381 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     4441 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     4501 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
     4561 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
     4621 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
     4681 ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc
     4741 atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
     4801 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
     4861 gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
     4921 ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct
     4981 gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct
     5041 caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca
     5101 ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg
     5161 atcatgcgca cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt
     5221 ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca
     5281 agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag
     5341 agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg
     5401 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     5461 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     5521 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5581 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     5641 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
     5701 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
     5761 tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc
     5821 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
     5881 agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
     5941 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
     6001 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
     6061 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
     6121 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
     6181 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
     6241 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
     6301 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
     6361 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
     6421 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
     6481 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
     6541 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
     6601 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
     6661 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
     6721 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca
     6781 aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
     6841 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
     6901 aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga
     6961 ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga
     7021 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat accatgggca
     7081 gcagccatca tcatcatcat cacagcagcg gcctggtgcc gcgcggcagc catatgtcac
     7141 ctatattagg atactggaaa atta
//
