LOCUS       sequence-382819-        7756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of Dmp68.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7756)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 7756)
  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: pETM-30-GST-Dmp68_B
FEATURES             Location/Qualifiers
     source          1..7756
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             94..111
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             130..780
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPP"
     CDS             817..837
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2621..2638
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2691..2709)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2705..2752
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2789..3244
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2876..2895)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3086..3107
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3336..4151)
                     /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(3336..4151)
                     /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     4059..4078
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4143..4162
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4273..4861
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4762..4781
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5015..5032
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5047..5186
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5112..5131)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5272..5294
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5288..5479)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6288..7370)
                     /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     7332..7351
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7371..7448)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7654..7673
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg aaacatcacc atcaccatca caacactagt
      121 agcaattcca tgtcccctat actaggttat tggaaaatta agggccttgt gcaacccact
      181 cgacttcttt tggaatatct tgaagaaaaa tatgaagagc atttgtatga gcgcgatgaa
      241 ggtgataaat ggcgaaacaa aaagtttgaa ttgggtttgg agtttcccaa tcttccttat
      301 tatattgatg gtgatgttaa attaacacag tctatggcca tcatacgtta tatagctgac
      361 aagcacaaca tgttgggtgg ttgtccaaaa gagcgtgcag agatttcaat gcttgaagga
      421 gcggttttgg atattagata cggtgtttcg agaattgcat atagtaaaga ctttgaaact
      481 ctcaaagttg attttcttag caagctacct gaaatgctga aaatgttcga agatcgttta
      541 tgtcataaaa catatttaaa tggtgatcat gtaacccatc ctgacttcat gttgtatgac
      601 gctcttgatg ttgttttata catggaccca atgtgcctgg atgcgttccc aaaattagtt
      661 tgttttaaaa aacgtattga agctatccca caaattgata agtacttgaa atccagcaag
      721 tatatagcat ggcctttgca gggctggcaa gccacgtttg gtggtggcga ccatcctcca
      781 actagtggat ctggtggtgg tggcggatgg atgagcgaga atctttattt tcagggcgcc
      841 atggcaccac acgatcgcga ctttggtcac agtggacgtg gcggtcgcgg aggcgatcga
      901 ggcggcgatg atcggcgagg aggcggcggc ggaggcaatc gcttcggtgg cggcggtggc
      961 ggtggcgatt accacggcat aaggaatgga cgcgtcgaga agcgacgcga tgaccgcgga
     1021 ggcggcaacc ggtttggagg tggtggtgga ttcggggatc gccgaggagg aggtggcggc
     1081 ggcagccaag acctccccat gcgtccggtc gacttctcca acctggctcc cttcaagaag
     1141 aacttttacc aggagcaccc taacgtagca aaccgatcgc cctacgaagt tcagaggtat
     1201 cgcgaagagc aggagatcac cgtgcgcgga caggtgccga accccatcca ggacttctcc
     1261 gaggtccatc tgcccgacta cgtcatgaag gagatccgcc gacagggcta caaggccccc
     1321 accgctatcc aagcgcaggg ctggcctatc gccatgagtg gctcgaactt cgtcggcatt
     1381 gccaagacgg gatccggaaa gaccctgggc tacatcctgc ccgctattgt ccacatcaac
     1441 aaccagcagc cgctgcagag gggcgacgga cctatcgccc tcgtgctggc ccccactcgg
     1501 gagctggccc aacagatcca gcaggtggcc accgaattcg gttcctcgtc gtatgttcgc
     1561 aacacctgcg tcttcggcgg cgccccgaag ggcggccaaa tgcgggatct gcagcgcggc
     1621 tgcgagatcg tcattgccac acccggacgt cttattgatt tcctctccgc cggctctact
     1681 aacctgaagc gttgcaccta cttggtgctg gacgaggccg atcgcatgtt agacatgggc
     1741 ttcgagcccc agatcagaaa gatcgtctca cagatccggc ccgaccgaca gacgctcatg
     1801 tggagcgcta cctggcccaa ggaggtcaag cagcttgccg aggacttttt gggcaactac
     1861 atccagatca acattggatc gctggagctg tccgccaatc acaacatccg ccaggtggtg
     1921 gacgtttgtg acgagttcag caaggaggag aaattgaaga ccctcctgtc agacatctac
     1981 gacaccagcg agagccccgg caagattatc atattcgtgg agacaaagcg acgcgtggac
     2041 aacctggtgc gcttcatccg cagcttcgga gtccgttgtg gagctattca cggtgacaag
     2101 tcgcaatcag aacgagactt tgtgctccgt gagttccgct cgggcaagtc caacattctg
     2161 gtggccaccg atgtggcggc ccgtggacta gacgtggacg gcatcaagta tgtcatcaac
     2221 tttgactacc cgcaaaacag cgaggactac atccatcgca tcggtcgcac aggacgatcc
     2281 aacacaaagg gcacctcttt cgccttcttc accaagaaca atgccaagca agccaaggcg
     2341 ctggtcgacg tactccgaga agctaatcag gaaatcaatc ctgccctgga aaatctggcc
     2401 cgcaactcgc gctacgacgg tggcggcgga cgctcccgtt atggaggcgg tggcggcggc
     2461 ggtcgcttcg gcggcggcgg cttcaagaag ggcagcctga gcaacggacg aggcttcgga
     2521 ggaggcggcg gcggcggcgg cgaaggcaga cactcgcgct tcgactaggg taccggatcc
     2581 gaattcgagc tccgtcgaca agcttgcggc cgcactcgag caccaccacc accaccactg
     2641 agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     2701 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     2761 aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     2821 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2881 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2941 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3001 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3061 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3121 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3181 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     3241 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     3301 tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc gagcatcaaa
     3361 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     3421 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     3481 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     3541 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt
     3601 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     3661 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     3721 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     3781 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     3841 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     3901 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     3961 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     4021 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     4081 tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga
     4141 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     4201 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4261 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4321 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4381 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     4441 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4501 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4561 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4621 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4681 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4741 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4801 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4861 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4921 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     4981 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     5041 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg
     5101 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     5161 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     5221 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     5281 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     5341 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     5401 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     5461 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     5521 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     5581 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     5641 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     5701 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     5761 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     5821 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     5881 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
     5941 cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg
     6001 gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc
     6061 cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg
     6121 gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca
     6181 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag
     6241 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
     6301 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6361 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
     6421 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
     6481 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
     6541 tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta
     6601 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
     6661 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
     6721 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
     6781 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
     6841 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
     6901 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
     6961 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
     7021 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
     7081 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
     7141 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
     7201 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
     7261 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
     7321 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
     7381 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
     7441 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     7501 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
     7561 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     7621 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     7681 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat
     7741 ctcgatcccg cgaaat
//
