LOCUS       sequence-382871-        6922 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 6922)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 6922)
  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-11-6His-Dmp68_B
FEATURES             Location/Qualifiers
     source          1..6922
                     /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             1787..1804
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1857..1875)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1871..1918
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1955..2410
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2042..2061)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2252..2273
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2502..3317)
                     /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(2502..3317)
                     /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     3225..3244
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3309..3328
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3439..4027
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3928..3947
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4181..4198
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4213..4352
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4278..4297)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4438..4460
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4454..4645)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5454..6536)
                     /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     6498..6517
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6537..6614)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6820..6839
                     /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 atggcaatga gtaatgctat taaccaaaat ggcacaggtc tagagcagca agttgctggt
      901 ctggacttga atggcggctc tgctgactac agcggcccca taaccagcaa gacttccact
      961 aactcggtca ccggtggtgt gtatgtgccc ccgcaccttc gtggtggtgg tggcaataac
     1021 aacgcagcgg acgcagagag ccagggccaa ggacagggcc agggtcaggg cttcgactcc
     1081 agatccggga atccgcgcca ggagactagg gaccctcaac agtcgcgcgg aggcggaggc
     1141 gaataccgca gaggcggcgg cggtggtggt cgcggcttca atcgccagag cggcgactac
     1201 ggttacggaa gcggcggcgg aggacgacgt ggaggcggtg gacgtttcga ggacaactac
     1261 aacggtggtg aattcgattc gcgtcgcggc ggtgactgga atcgcagcgg cggaggaggt
     1321 cgaggatttg gacgcggacc atcgtaccgc ggcggtggcg gcggaagcgg tagcaacctc
     1381 aacgagcaga ctgccgagga tggccaggca cagcagcagc agcagccgcg caacgatcgt
     1441 tggcaggagc cagaacgtcc tgccggattc gatggcagcg agggcggaca gtctgccggc
     1501 ggcaacagaa gctacaacaa tcgcggcgag cgcggaggcg gcggttacaa cagccgctgg
     1561 aaggagggcg gcggctcaaa cgtcgactac acaaagctgg gcgcccgaga tgagcgcctg
     1621 gaggtggagc tgttcggcgt gggcaatacg ggaatcaatt ttgacaaata cgaggatata
     1681 cccgtggagg cgacgggcca gaatgtgccc ccgaacatca catcgttcga ttagggtacc
     1741 ggatccgaat tcgagctccg tcgacaagct tgcggccgca ctcgagcacc accaccacca
     1801 ccactgagat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     1861 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     1921 gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     1981 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2041 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2101 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2161 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2221 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2281 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2341 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     2401 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     2461 tctaaataca ttcaaatatg tatccgctca tgaattaatt cttagaaaaa ctcatcgagc
     2521 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     2581 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     2641 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     2701 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     2761 aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     2821 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
     2881 acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
     2941 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
     3001 gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
     3061 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
     3121 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
     3181 ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     3241 tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca agacgtttcc
     3301 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt
     3361 gttcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3421 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3481 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3541 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     3601 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3661 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3721 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     3781 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     3841 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     3901 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     3961 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4021 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4081 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     4141 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     4201 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4261 caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     4321 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     4381 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     4441 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     4501 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     4561 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     4621 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
     4681 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
     4741 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
     4801 agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac
     4861 agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact
     4921 tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg
     4981 tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct
     5041 gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca
     5101 tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg
     5161 tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg
     5221 acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg
     5281 ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga
     5341 tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg
     5401 gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     5461 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5521 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc
     5581 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
     5641 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     5701 ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac
     5761 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     5821 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     5881 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     5941 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     6001 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
     6061 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     6121 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     6181 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
     6241 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     6301 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     6361 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     6421 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
     6481 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
     6541 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc
     6601 cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca
     6661 gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga
     6721 gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
     6781 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
     6841 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat
     6901 cgagatctcg atcccgcgaa at
//
