LOCUS       sequence-360748-        7258 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial Expression of DmGe1_1220-1354.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7258)
  AUTHORS   Jinek M, Eulalio A, Lingel A, Helms S, Conti E, Izaurralde E
  TITLE     The C-terminal region of Ge-1 presents conserved structural features
            required for P-body localization.
  JOURNAL   RNA. 2008 Oct . 14(10):1991-8.
  PUBMED    18755833
REFERENCE   2  (bases 1 to 7258)
  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: "RNA. 2008 
            Oct . 14(10):1991-8."
COMMENT     Sequence Label: pETM-60-NusA-DmGe1_1220-1354_G
FEATURES             Location/Qualifiers
     source          1..7258
                     /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             2123..2140
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2193..2211)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2207..2254
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2291..2746
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2378..2397)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2588..2609
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2838..3653)
                     /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(2838..3653)
                     /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     3561..3580
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3645..3664
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3775..4363
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4264..4283
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4517..4534
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4549..4688
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4614..4633)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4774..4796
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4790..4981)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5790..6872)
                     /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     6834..6853
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6873..6950)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7156..7175
                     /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 gcgccatggc tacgcctgca ccaccttacg acctcaggga cagcattaag
     1681 cagcttttga tggctggaca aatcaacaag gccttccatc aggctctatt ggccaacgac
     1741 ttgggcctag tggagttcac tctccgacac acggacagca accaggcatt cgcaccagag
     1801 ggatgcagac tggagcaaaa ggttctactg tcgctcattc agcaaatatc cgctgacatg
     1861 accaatcaca atgagcttaa gcagagatat ctcaacgagg cactgttggc catcaatatg
     1921 gcagatccca ttacccgcga gcacgctcct aaagtgctca ccgagctgta tcgcaattgt
     1981 cagcagttca ttaagaacag ccccaagaat tcgcagttca gcaacgtccg tctgctgatg
     2041 aaggcaatca taacgtaccg cgatcagcta aaatgaggat ccgaattcga gctccgtcga
     2101 caagcttgcg gccgcactcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa
     2161 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
     2221 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt
     2281 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2341 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2401 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2461 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2521 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2581 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2641 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2701 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     2761 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     2821 cgctcatgaa ttaattctta gaaaaactca tcgagcatca aatgaaactg caatttattc
     2881 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
     2941 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
     3001 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     3061 tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag
     3121 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     3181 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa
     3241 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     3301 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg
     3361 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     3421 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     3481 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc
     3541 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     3601 ttggaattta atcgcggcct agagcaagac gtttcccgtt gaatatggct cataacaccc
     3661 cttgtattac tgtttatgta agcagacagt tttattgttc atgaccaaaa tcccttaacg
     3721 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3781 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3841 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3901 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     3961 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4021 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4081 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4141 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4201 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4261 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     4321 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     4381 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     4441 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     4501 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     4561 ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc
     4621 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     4681 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     4741 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     4801 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     4861 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     4921 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     4981 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag
     5041 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg
     5101 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat
     5161 gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga
     5221 tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa
     5281 cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt
     5341 cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc
     5401 agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc
     5461 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt
     5521 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc
     5581 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt
     5641 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
     5701 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     5761 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5821 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5881 gccagggtgg tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc
     5941 tggccctgag agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc
     6001 tgtttgatgg tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc
     6061 actaccgaga tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc
     6121 agcgccatct gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt
     6181 tgcatggttt gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc
     6241 tgaatttgat tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca
     6301 gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc
     6361 acgcccagtc gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca
     6421 gagacatcaa gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc
     6481 tggtcatcca gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc
     6541 accgccgctt tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca
     6601 cccagttgat cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc
     6661 agactggagg tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg
     6721 cggttgggaa tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca
     6781 gaaacgtggc tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac
     6841 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
     6901 gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg
     6961 acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt
     7021 gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc
     7081 cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc
     7141 ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc
     7201 ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc cgcgaaat
//
