LOCUS       sequence-489470-        6922 bp    DNA     circular SYN 16-OCT-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 Oct 16, 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"
     CDS             98..118
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1068..1085
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1138..1156)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1152..1199
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1236..1691
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1323..1342)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1533..1554
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1783..2598)
                     /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(1783..2598)
                     /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     2506..2525
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2590..2609
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2720..3308
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3209..3228
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3462..3479
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3494..3633
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3559..3578)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3719..3741
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3735..3926)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4735..5817)
                     /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     5779..5798
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5818..5895)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6101..6120
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6204..6223
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6204..6222
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6223..6247
                     /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             6262..6284
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             6297..6314
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(6333..6922,1..61)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPP"
ORIGIN
        1 gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg accatcctcc
       61 aactagtgga tctggtggtg gtggcggatg gatgagcgag aatctttatt ttcagggcgc
      121 catggcaatg agtaatgcta ttaaccaaaa tggcacaggt ctagagcagc aagttgctgg
      181 tctggacttg aatggcggct ctgctgacta cagcggcccc ataaccagca agacttccac
      241 taactcggtc accggtggtg tgtatgtgcc cccgcacctt cgtggtggtg gtggcaataa
      301 caacgcagcg gacgcagaga gccagggcca aggacagggc cagggtcagg gcttcgactc
      361 cagatccggg aatccgcgcc aggagactag ggaccctcaa cagtcgcgcg gaggcggagg
      421 cgaataccgc agaggcggcg gcggtggtgg tcgcggcttc aatcgccaga gcggcgacta
      481 cggttacgga agcggcggcg gaggacgacg tggaggcggt ggacgtttcg aggacaacta
      541 caacggtggt gaattcgatt cgcgtcgcgg cggtgactgg aatcgcagcg gcggaggagg
      601 tcgaggattt ggacgcggac catcgtaccg cggcggtggc ggcggaagcg gtagcaacct
      661 caacgagcag actgccgagg atggccaggc acagcagcag cagcagccgc gcaacgatcg
      721 ttggcaggag ccagaacgtc ctgccggatt cgatggcagc gagggcggac agtctgccgg
      781 cggcaacaga agctacaaca atcgcggcga gcgcggaggc ggcggttaca acagccgctg
      841 gaaggagggc ggcggctcaa acgtcgacta cacaaagctg ggcgcccgag atgagcgcct
      901 ggaggtggag ctgttcggcg tgggcaatac gggaatcaat tttgacaaat acgaggatat
      961 acccgtggag gcgacgggcc agaatgtgcc cccgaacatc acatcgttcg attagggtac
     1021 cggatccgaa ttcgagctcc gtcgacaagc ttgcggccgc actcgagcac caccaccacc
     1081 accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     1141 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     1201 tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag
     1261 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     1321 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     1381 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     1441 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     1501 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     1561 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     1621 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     1681 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1741 ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa actcatcgag
     1801 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     1861 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     1921 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     1981 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     2041 caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     2101 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa
     2161 tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     2221 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     2281 tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     2341 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     2401 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     2461 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     2521 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc
     2581 ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag acagttttat
     2641 tgttcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     2701 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2761 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2821 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     2881 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2941 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3001 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3061 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3121 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3181 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3241 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3301 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3361 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     3421 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     3481 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3541 gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     3601 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     3661 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     3721 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     3781 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     3841 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     3901 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     3961 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     4021 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     4081 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     4141 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     4201 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     4261 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     4321 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     4381 atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg
     4441 gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc
     4501 gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc
     4561 gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg
     4621 atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     4681 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     4741 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4801 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     4861 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     4921 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     4981 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     5041 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     5101 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     5161 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     5221 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     5281 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     5341 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     5401 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     5461 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     5521 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     5581 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     5641 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     5701 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     5761 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     5821 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg
     5881 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc
     5941 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg
     6001 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     6061 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     6121 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
     6181 tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
     6241 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc atgaaacatc
     6301 accatcacca tcacaacact agtagcaatt ccatgtcccc tatactaggt tattggaaaa
     6361 ttaagggcct tgtgcaaccc actcgacttc ttttggaata tcttgaagaa aaatatgaag
     6421 agcatttgta tgagcgcgat gaaggtgata aatggcgaaa caaaaagttt gaattgggtt
     6481 tggagtttcc caatcttcct tattatattg atggtgatgt taaattaaca cagtctatgg
     6541 ccatcatacg ttatatagct gacaagcaca acatgttggg tggttgtcca aaagagcgtg
     6601 cagagatttc aatgcttgaa ggagcggttt tggatattag atacggtgtt tcgagaattg
     6661 catatagtaa agactttgaa actctcaaag ttgattttct tagcaagcta cctgaaatgc
     6721 tgaaaatgtt cgaagatcgt ttatgtcata aaacatattt aaatggtgat catgtaaccc
     6781 atcctgactt catgttgtat gacgctcttg atgttgtttt atacatggac ccaatgtgcc
     6841 tggatgcgtt cccaaaatta gtttgtttta aaaaacgtat tgaagctatc ccacaaattg
     6901 ataagtactt gaaatccagc aa
//
