LOCUS       sequence-360155-        7162 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmEDC3_331-440.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7162)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 7162)
  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-60-NusA-DmEDC3_331-440_H
FEATURES             Location/Qualifiers
     source          1..7162
                     /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             88..1569
                     /codon_start=1
                     /gene="nusA"
                     /product="transcription elongation protein (N utilization 
                     substance protein A)"
                     /label=NusA
                     /note="highly soluble in E. coli"
                     /translation="MKEILAVVEAVSNEKALPREKIFEALESALATATKKKYEQEIDVR
                     VQIDRKSGDFDTFRRWLVVDEVTQPTKEITLEAARYEDESLNLGDYVEDQIESVTFDRI
                     TTQTAKQVIVQKVREAERAMVVDQFREHEGEIITGVVKKVNRDNISLDLGNNAEAVILR
                     EDMLPRENFRPGDRVRGVLYSVRPEARGAQLFVTRSKPEMLIELFRIEVPEIGEEVIEI
                     KAAARDPGSRAKIAVKTNDKRIDPVGACVGMRGARVQAVSTELGGERIDIVLWDDNPAQ
                     FVINAMAPADVASIVVDEDKHTMDIAVEAGNLAQAIGRNGQNVRLASQLSGWELNVMTV
                     DDLQAKHQAEAHAAIDTFTKYLDIDEDFATVLVEEGFSTLEELAYVPMKELLEIEGLDE
                     PTVEALRERAKNALATIAQAQEESLGDNKPADDLLNLEGVDRDLAFKLAARGVCTLEDL
                     AEQGIDDLADIEGLTDEKAGALIMAARNICWFGDEA"
     CDS             1585..1602
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1612..1632
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2027..2044
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2097..2115)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2111..2158
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2195..2650
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2282..2301)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2492..2513
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2742..3557)
                     /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(2742..3557)
                     /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     3465..3484
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3549..3568
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3679..4267
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4168..4187
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4421..4438
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4453..4592
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4518..4537)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4678..4700
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4694..4885)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5694..6776)
                     /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     6738..6757
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6777..6854)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7060..7079
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg aaagaaattt tggctgtagt tgaagccgta
      121 tccaatgaaa aggcgctacc tcgcgagaag attttcgaag cattggaaag cgcgctggcg
      181 acagcaacaa agaaaaaata tgaacaagag atcgacgtcc gcgtacagat cgatcgcaaa
      241 agcggtgatt ttgacacttt ccgtcgctgg ttagttgttg atgaagtcac ccagccgacc
      301 aaggaaatca cccttgaagc cgcacgttat gaagatgaaa gcctgaacct gggcgattac
      361 gttgaagatc agattgagtc tgttaccttt gaccgtatca ctacccagac ggcaaaacag
      421 gttatcgtgc agaaagtgcg tgaagccgaa cgtgcgatgg tggttgatca gttccgtgaa
      481 cacgaaggtg aaatcatcac cggcgtggtg aaaaaagtaa accgcgacaa catctctctg
      541 gatctgggca acaacgctga agccgtgatc ctgcgcgaag atatgctgcc gcgtgaaaac
      601 ttccgccctg gcgaccgcgt tcgtggcgtg ctctattccg ttcgcccgga agcgcgtggc
      661 gcgcaactgt tcgtcactcg ttccaagccg gaaatgctga tcgaactgtt ccgtattgaa
      721 gtgccagaaa tcggcgaaga agtgattgaa attaaagcag cggctcgcga tccgggttct
      781 cgtgcgaaaa tcgcggtgaa aaccaacgat aaacgtatcg atccggtagg tgcttgcgta
      841 ggtatgcgtg gcgcgcgtgt tcaggcggtg tctactgaac tgggtggcga gcgtatcgat
      901 atcgtcctgt gggatgataa cccggcgcag ttcgtgatta acgcaatggc accggcagac
      961 gttgcttcta tcgtggtgga tgaagataaa cacactatgg acatcgccgt tgaagccggt
     1021 aatctggcgc aggcgattgg ccgtaacggt cagaacgtgc gtctggcttc gcaactgagc
     1081 ggttgggaac tcaacgtgat gaccgttgac gacctgcaag ctaagcatca ggcggaagcg
     1141 cacgcagcga tcgacacctt caccaaatat ctcgacatcg acgaagactt cgcgactgtt
     1201 ctggtagaag aaggcttctc gacgctggaa gaattggcct atgtgccgat gaaagagctg
     1261 ttggaaatcg aaggccttga tgagccgacc gttgaagcac tgcgcgagcg tgctaaaaat
     1321 gcactggcca ccattgcaca ggcccaggaa gaaagcctcg gtgataacaa accggctgac
     1381 gatctgctga accttgaagg ggtagatcgt gatttggcat tcaaactggc cgcccgtggc
     1441 gtttgtacgc tggaagatct cgccgaacag ggcattgatg atctggctga tatcgaaggg
     1501 ttgaccgacg aaaaagccgg agcactgatt atggctgccc gtaatatttg ctggttcggt
     1561 gacgaagcga ctagtggttc tggtcatcac catcaccatc actccgcggg tgagaatctt
     1621 tattttcagg gcgccatggt aacttttggc agcgaggcgg acgatccgct gatccacgag
     1681 gatttcgatt ttgagggcaa tctagcgctg ttcgacaagc aggcaatttg ggacgacatc
     1741 gagtcgacga cccaaaaacc agatgtggtg cggcacattg tgaacaatca tcatcataag
     1801 ccagagcaga agtatcggca cgacgagaac atcctggcca gcaaaccact acaattgcgc
     1861 cagatcgaaa gcatgttcgg cggcagccag gactttgtga cggacgacgg tctcattata
     1921 ccgacgattc cggcttacgt tcggaataaa atcgagataa gtgccgataa gtagggtacc
     1981 ggatccgaat tcgagctccg tcgacaagct tgcggccgca ctcgagcacc accaccacca
     2041 ccactgagat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     2101 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     2161 gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     2221 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2281 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2341 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2401 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     2461 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     2521 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2581 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     2641 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     2701 tctaaataca ttcaaatatg tatccgctca tgaattaatt cttagaaaaa ctcatcgagc
     2761 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     2821 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     2881 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     2941 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     3001 aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     3061 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
     3121 acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
     3181 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
     3241 gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
     3301 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
     3361 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
     3421 ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     3481 tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca agacgtttcc
     3541 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt
     3601 gttcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     3661 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     3721 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     3781 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     3841 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     3901 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     3961 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     4021 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     4081 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     4141 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     4201 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     4261 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     4321 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     4381 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     4441 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4501 caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     4561 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     4621 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     4681 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     4741 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     4801 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     4861 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
     4921 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
     4981 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
     5041 agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac
     5101 agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact
     5161 tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg
     5221 tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct
     5281 gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca
     5341 tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg
     5401 tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg
     5461 acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg
     5521 ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga
     5581 tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg
     5641 gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     5701 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5761 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc
     5821 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
     5881 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     5941 ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac
     6001 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     6061 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     6121 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     6181 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     6241 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
     6301 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     6361 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     6421 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
     6481 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     6541 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     6601 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     6661 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
     6721 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
     6781 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc
     6841 cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaagca
     6901 gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga
     6961 gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
     7021 gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc
     7081 gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat
     7141 cgagatctcg atcccgcgaa at
//
