LOCUS       sequence-386314-        7353 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  protein expression of the GST-tagged catalytic domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7353)
  TITLE     Knapp lab plasmids
REFERENCE   2  (bases 1 to 7353)
  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: pGTVL2-SENP6
FEATURES             Location/Qualifiers
     source          1..7353
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     41..63
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(57..248)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1057..2139)
                     /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     2101..2120
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2140..2217)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2423..2442
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2526..2545
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2526..2544
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2545..2569
                     /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             2584..2606
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2617..2634
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2641..3294
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGADHPPK"
     CDS             3322..3342
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             4743..4760
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4813..4831)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4827..4874
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4911..5366
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4998..5017)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5208..5229
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5458..6273)
                     /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(5458..6273)
                     /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     6181..6200
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6265..6284
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6395..6983
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6884..6903
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7137..7154
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7169..7308
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7234..7253)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
       61 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
      121 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
      181 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
      241 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
      301 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
      361 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
      421 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
      481 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
      541 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
      601 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
      661 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc
      721 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
      781 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
      841 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
      901 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
      961 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     1021 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     1081 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     1141 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     1201 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     1261 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     1321 gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     1381 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     1441 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     1501 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     1561 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     1621 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     1681 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     1741 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     1801 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     1861 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     1921 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     1981 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     2041 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     2101 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     2161 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     2221 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
     2281 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
     2341 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     2401 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     2461 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc
     2521 gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc ctctagaaat
     2581 aattttgttt aactttaaga aggagatata catatgcacc atcatcatca tcattcttct
     2641 atgtccccta tactaggtta ttggaaaatt aagggccttg tgcaacccac tcgacttctt
     2701 ttggaatatc ttgaagaaaa atatgaagag catttgtatg agcgcgatga aggtgataaa
     2761 tggcgaaaca aaaagtttga attgggtttg gagtttccca atcttcctta ttatattgat
     2821 ggtgatgtta aattaacaca gtctatggcc atcatacgtt atatagctga caagcacaac
     2881 atgttgggtg gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg agcggttttg
     2941 gatattagat acggtgtttc gagaattgca tatagtaaag actttgaaac tctcaaagtt
     3001 gattttctta gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt atgtcataaa
     3061 acatatttaa atggtgatca tgtaacccat cctgacttca tgttgtatga cgctcttgat
     3121 gttgttttat acatggaccc aatgtgcctg gatgcgttcc caaaattagt ttgttttaaa
     3181 aaacgtattg aagctatccc acaaattgat aagtacttga aatccagcaa gtatatagca
     3241 tggcctttgc agggctggca agccacgttt ggtggtgccg accatcctcc aaaatcgagc
     3301 tcaggtgtag atctgggtac cgagaacctg tacttccaat ccatggaaaa actgattgtg
     3361 tatccgccgc cgccggccaa aggcggtatc agcgtgacca atgaagatct gcactgcctg
     3421 aacgagggtg aatttctgaa tgacgtgatt atcgatttct atctgaaata cctggttctg
     3481 gaaaaactga aaaaagaaga cgccgatcgc atccatatct tttcatcgtt tttctataaa
     3541 cgcctgaacc agcgtgaacg tcgcaaccac gaaaccacga atctgagcat tcagcaaaaa
     3601 cgccatggcc gtgtgaaaac ctggacgcgt cacgttgaca tcttcgaaaa agatttcatc
     3661 ttcgttccgc tgaatgaagc ggcccattgg tttctggcag tggtttgctt cccgggtctg
     3721 gaaaaaccga aatatgaacc gaacccgcat taccacgaaa atgctgtcat tcagaaatgc
     3781 tcaaccgtgg aagattcgtg tatcagctct agtgcgagcg aaatggaatc ttgcagtcag
     3841 aactcctcag ccaaaccggt tattaagaaa atgctgaaca aaaaacattg tattgcggtc
     3901 atcgactcta atccgggcca ggaagaaagt gatccgcgct acaaacgtaa catttgttcc
     3961 gttaaatact cagtgaagaa aattaaccac accgcgagcg aaaacgaaga attcaacaag
     4021 ggtgaatcca cctcacagaa agtggccgat cgcacgaaat cggaaaacgg cctgcaaaat
     4081 gaaagcctgt cgagcaccca tcacacggac ggtctgtcga aaattcgtct gaactactcg
     4141 gatgaaagcc cggaagccgg caaaatgctg gaagacgaac tggttgattt ttcagaagac
     4201 caggataacc aagacgattc tagtgacgat ggcttcctgg cagacgataa ttgctcctca
     4261 gaaattggcc agtggcatct gaaaccgacc atttgcaaac aaccgtgtat cctgctgatg
     4321 gattctctgc gcggcccgtc gcgtagcaat gtcgtgaaaa tcctgcgcga atatctggaa
     4381 gtggaatggg aagttaaaaa aggcagtaaa cgttctttca gtaaagatgt catgaaaggt
     4441 tccaacccga aagtgccgca gcaaaacaat ttctcagatt gtggtgttta tgtcctgcag
     4501 tacgtggaaa gctttttcga aaatccgatt ctgagttttg aactgccgat gaacctggcg
     4561 aattggtttc cgccgccgcg tatgcgcacc aaacgcgaag aaatccgtaa tattatcctg
     4621 aaactgcagg aagaccaaag caaagaaaaa cgtaaacaca aagatacgta ttcaaccgaa
     4681 gcttaaaggt ggatacggat ccgaattcga gctccgtcga caagcttgcg gccgcactcg
     4741 agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt
     4801 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
     4861 tgaggggttt tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg
     4921 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4981 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5041 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5101 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     5161 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5221 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     5281 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     5341 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     5401 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgaa ttaattctta
     5461 gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc
     5521 atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag
     5581 gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat
     5641 taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga
     5701 atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc
     5761 attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc
     5821 ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg
     5881 caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc
     5941 ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc
     6001 aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag
     6061 tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa
     6121 ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt
     6181 atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta atcgcggcct
     6241 agagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta
     6301 agcagacagt tttattgttc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6361 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6421 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6481 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6541 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6601 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6661 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6721 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6781 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6841 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6901 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6961 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     7021 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     7081 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     7141 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     7201 gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     7261 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     7321 ccaacacccg ctgacgcgcc ctgacgggct tgt
//
