LOCUS       sequence-111024-        5191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  His-GST-Smt3-NpuDnaEN102-CBD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5191)
  AUTHORS   Guerrero F, Ciragan A, Iwai H
  TITLE     Tandem SUMO fusion vectors for improving soluble protein expression 
            and purification.
  JOURNAL   Protein Expr Purif. 2015 Aug 20. pii: S1046-5928(15)30039-5. doi: 
            10.1016/j.pep.2015.08.019.
  PUBMED    26297996
REFERENCE   2  (bases 1 to 5191)
  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"; journalName: "Protein 
            Expr Purif. 2015 Aug 20. pii: S1046-5928(15)30039-5. doi: 
            10.1016/j.pep.2015.08.019."
COMMENT     Sequence Label: pLJSRSF3
FEATURES             Location/Qualifiers
     source          1..5191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             121..771
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="SPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKF
                     ELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYG
                     VSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLY
                     MDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     729..751
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             775..1065
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="SDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLR
                     RLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIETHREQIGG"
     CDS             1561..1716
                     /codon_start=1
                     /gene="Bacillus circulans chiA"
                     /product="chitin binding domain from chitinase A1 (Watanabe
                     et al., 1994)"
                     /label=CBD
                     /translation="TTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNV
                     PALWQLQ"
     CDS             1745..1789
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1827..1845)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1841..1888
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2118..2933)
                     /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(2118..2933)
                     /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     2841..2860
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2934..3025)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3041..3790)
                     /direction=LEFT
                     /label=RSF ori
                     /note="Plasmids containing the RSF 1030 origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     primer_bind     complement(3051..3068)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3291..3310)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3986..5068)
                     /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     5030..5049
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5069..5146)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg ggcagcagcc atcatcatca tcatcacggc
      121 tcccctatac taggttattg gaaaattaag ggccttgtgc aacccactcg acttcttttg
      181 gaatatcttg aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg tgataaatgg
      241 cgaaacaaaa agtttgaatt gggtttggag tttcccaatc ttccttatta tattgatggt
      301 gatgttaaat taacacagtc tatggccatc atacgttata tagctgacaa gcacaacatg
      361 ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc ggttttggat
      421 attagatacg gtgtttcgag aattgcatat agtaaagact ttgaaactct caaagttgat
      481 tttcttagca agctacctga aatgctgaaa atgttcgaag atcgtttatg tcataaaaca
      541 tatttaaatg gtgatcatgt aacccatcct gacttcatgt tgtatgacgc tcttgatgtt
      601 gttttataca tggacccaat gtgcctggat gcgttcccaa aattagtttg ttttaaaaaa
      661 cgtattgaag ctatcccaca aattgataag tacttgaaat ccagcaagta tatagcatgg
      721 cctttgcagg gctggcaagc cacgtttggt ggtggcgacc atcctccaaa aactagtgac
      781 tcagaagtca atcaagaagc taagccagag gtcaagccag aagtcaagcc tgagactcac
      841 atcaatttaa aggtgtccga tggatcttca gaaatcttct ttaagatcaa aaagaccact
      901 cctttaagaa ggctgatgga agcgttcgct aaaagacagg gtaaggaaat ggactcctta
      961 agattcttgt acgacggtat tagaattcaa gctgatcaga cccctgaaga tttggacatg
     1021 gaggataacg atattattga gactcacaga gaacagattg gtggatccca tatgcagtac
     1081 aaacttatcc tgaacggtaa aaccctgaaa ggtgaaacca ccaccgaagc tgttgacgct
     1141 gctaccgcgg aaaaagtttt caaacagtac gctaacgaca acggtgttga cggtgaatgg
     1201 acctacgacg acgctaccaa aacctacacc gtaacggaag gatcctgttt aagctatgaa
     1261 acggaaatat tgacagtaga atatggatta ttaccgattg gtaaaattgt agaaaagcgc
     1321 atcgaatgta ctgtttatag cgttgataat aatggaaata tttatacaca acctgtagca
     1381 caatggcacg atcgcggaga acaagaggtg tttgagtatt gtttggaaga tggttcattg
     1441 attcgggcaa caaaagacca taagtttatg actgttgatg gtcaaatgtt gccaattgat
     1501 gaaatatttg aacgtgaatt ggatttgatg cgggttgata atttgccgaa cgaattcggt
     1561 accacaaatc ctggtgtatc cgcttggcag gtcaacacag cttatactgc gggacaattg
     1621 gtcacatata acggcaagac gtataaatgt ttgcagcccc acacctcctt ggcaggatgg
     1681 gaaccatcca acgttcctgc cttgtggcag cttcaataag cttgcggccg cactcgagtc
     1741 tggtaaagaa accgctgctg cgaaatttga acgccagcac atggactcgt ctactagcgc
     1801 agcttaatta acctaggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     1861 ctctaaacgg gtcttgaggg gttttttgct gaaacctcag gcatttgaga agcacacggt
     1921 cacactgctt ccggtagtca ataaaccggt aaaccagcaa tagacataag cggctattta
     1981 acgaccctgc cctgaaccga cgacaagctg acgaccgggt ctccgcaagt ggcacttttc
     2041 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2101 cgctcatgaa ttaattctta gaaaaactca tcgagcatca aatgaaactg caatttattc
     2161 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
     2221 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
     2281 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     2341 tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag
     2401 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     2461 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc ggtcgctgtt aaaaggacaa
     2521 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     2581 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg
     2641 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     2701 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     2761 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc
     2821 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     2881 ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct catactcttc
     2941 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3001 gaatgtattt agaaaaataa acaaataggc atgcagcgct cttccgcttc ctcgctcact
     3061 gactcgctac gctcggtcgt tcgactgcgg cgagcggtgt cagctcactc aaaagcggta
     3121 atacggttat ccacagaatc aggggataaa gccggaaaga acatgtgagc aaaaagcaaa
     3181 gcaccggaag aagccaacgc cgcaggcgtt tttccatagg ctccgccccc ctgacgagca
     3241 tcacaaaaat cgacgctcaa gccagaggtg gcgaaacccg acaggactat aaagatacca
     3301 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3361 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgttg
     3421 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3481 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3541 cgacttatcg ccactggcag cagccattgg taactgattt agaggacttt gtcttgaagt
     3601 tatgcacctg ttaaggctaa actgaaagaa cagattttgg tgagtgcggt cctccaaccc
     3661 acttaccttg gttcaaagag ttggtagctc agcgaacctt gagaaaacca ccgttggtag
     3721 cggtggtttt tctttattta tgagatgatg aatcaatcgg tctatcaagt caacgaacag
     3781 ctattccgtt actctagatt tcagtgcaat ttatctcttc aaatgtagca cctgaagtca
     3841 gccccatacg atataagttg taattctcat gttagtcatg ccccgcgccc accggaagga
     3901 gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag
     3961 ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     4021 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca
     4081 gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc
     4141 cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt
     4201 tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta
     4261 ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg
     4321 ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca
     4381 tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa
     4441 tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac
     4501 ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc
     4561 ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga
     4621 catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt
     4681 catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg
     4741 ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca
     4801 gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac
     4861 tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt
     4921 tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa
     4981 cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg
     5041 cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc
     5101 gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc
     5161 tctcccttat gcgactcctg cattaggaaa t
//
