LOCUS       sequence-430165-        5949 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the synthetic glutathionine S-transferase with N-terminal 
            His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5949)
  TITLE     Shanbhag lab plasmids
REFERENCE   2  (bases 1 to 5949)
  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
FEATURES             Location/Qualifiers
     source          1..5949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     331..350
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(370..447)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     653..672
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     756..775
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        756..774
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    775..799
                     /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             814..836
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             855..872
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             882..899
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             903..1559
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             1569..1586
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1639..1657)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1653..1700
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1737..2192
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1824..1843)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2034..2055
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2284..3099)
                     /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(2284..3099)
                     /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     3007..3026
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3091..3110
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3221..3809
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3710..3729
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3963..3980
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3995..4134
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4060..4079)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4220..4242
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4236..4427)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5236..5949,1..369))
                     /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"
ORIGIN
        1 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
       61 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
      121 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
      181 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
      241 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
      301 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
      361 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg
      421 aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct
      481 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
      541 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
      601 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
      661 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
      721 cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta taggggaatt
      781 gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga aggagatata
      841 ccatgggcag cagccatcat catcatcatc acagcagcgg cctggtgccg cgcggcagcc
      901 atatgtctcc aattctggga tattggaaaa ttaagggtct cgtccaaccg acgcgtcttc
      961 ttctggaata ccttgaggaa aagtatgaag agcacttgta cgaacgcgat gagggcgata
     1021 aatggcgtaa caaaaaattc gaacttgggc tcgagttccc taatttgccg tattatatcg
     1081 atggggatgt gaaacttacc caatcaatgg ccataatccg ctacattgcg gataaacata
     1141 atatgttagg tggttgccca aaggagcggg cagaaatcag tatgttagaa ggcgccgtac
     1201 ttgacatccg ttatggtgtt tctagaatcg catattcaaa agactttgaa actcttaaag
     1261 tggactttct tagtaagctc cctgaaatgc tgaaaatgtt tgaggaccgc ttatgccata
     1321 aaacctactt aaatggcgat catgtaactc acccagattt tatgctgtac gatgcgttag
     1381 atgtcgtgtt atacatggac cccatgtgcc tggatgcctt tcccaaatta gtgtgcttca
     1441 agaaacgtat tgaggccata ccgcagatcg acaaatatct caaaagctct aaatatatcg
     1501 cttggccgct tcagggttgg caggctacat ttggtggcgg cgatcatccg ccgaagtgat
     1561 aactcgagca ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag
     1621 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     1681 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc
     1741 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     1801 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     1861 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     1921 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     1981 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2041 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2101 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2161 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     2221 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattaa
     2281 ttcttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
     2341 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     2401 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     2461 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     2521 gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg
     2581 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     2641 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     2701 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
     2761 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
     2821 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     2881 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     2941 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
     3001 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     3061 cggcctagag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
     3121 tatgtaagca gacagtttta ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3181 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3241 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3301 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3361 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3421 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3481 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3541 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3601 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3661 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3721 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3781 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3841 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3901 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3961 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     4021 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     4081 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     4141 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4201 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4261 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     4321 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     4381 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     4441 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     4501 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     4561 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     4621 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     4681 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     4741 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     4801 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     4861 ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc
     4921 tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc
     4981 aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc
     5041 tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag
     5101 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg
     5161 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     5221 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5281 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     5341 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     5401 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     5461 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc
     5521 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     5581 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     5641 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     5701 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     5761 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     5821 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     5881 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     5941 atagttaat
//
