LOCUS       sequence-223540-        6871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminus of E. coli NusG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6871)
  AUTHORS   Lawson MR, Ma W, Bellecourt MJ, Artsimovitch I, Martin A, Landick R,
            Schulten K, Berger JM
  TITLE     Mechanism for the Regulated Control of Bacterial Transcription 
            Termination by a Universal Adaptor Protein.
  JOURNAL   Mol Cell. 2018 Sep 20;71(6):911-922.e4. doi: 
            10.1016/j.molcel.2018.07.014. Epub 2018 Aug 16.
  PUBMED    30122535
REFERENCE   2  (bases 1 to 6871)
  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"; doi: 
            "10.1016/j.molcel.2018.07.014"; journalName: "Mol Cell"; date: 
            "2018-09-20- 20"; volume: "71"; issue: "6"; pages: "911-922.e4"
COMMENT     Sequence Label: EcNusGNTD
FEATURES             Location/Qualifiers
     source          1..6871
                     /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             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..1227
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1201..1224
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1282..1302
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1735..1752
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1805..1823)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1819..1866
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1903..2358
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1990..2009)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2200..2221
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2451..3266)
                     /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(2451..3266)
                     /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     3174..3193
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3258..3277
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3388..3976
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3877..3896
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4130..4147
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4162..4301
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4227..4246)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4387..4409
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4403..4594)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5403..6485)
                     /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     6447..6466
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6486..6563)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6769..6788
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggttcttctc accatcacca tcaccatggt
      121 tcttctatga aaatcgaaga aggtaaactg gtaatctgga ttaacggcga taaaggctat
      181 aacggtctcg ctgaagtcgg taagaaattc gagaaagata ccggaattaa agtcaccgtt
      241 gagcatccgg ataaactgga agagaaattc ccacaggttg cggcaactgg cgatggccct
      301 gacattatct tctgggcaca cgaccgcttt ggtggctacg ctcaatctgg cctgttggct
      361 gaaatcaccc cggacaaagc gttccaggac aagctgtatc cgtttacctg ggatgccgta
      421 cgttacaacg gcaagctgat tgcttacccg atcgctgttg aagcgttatc gctgatttat
      481 aacaaagatc tgctgccgaa cccgccaaaa acctgggaag agatcccggc gctggataaa
      541 gaactgaaag cgaaaggtaa gagcgcgctg atgttcaacc tgcaagaacc gtacttcacc
      601 tggccgctga ttgctgctga cgggggttat gcgttcaagt atgaaaacgg caagtacgac
      661 attaaagacg tgggcgtgga taacgctggc gcgaaagcgg gtctgacctt cctggttgac
      721 ctgattaaaa acaaacacat gaatgcagac accgattact ccatcgcaga agctgccttt
      781 aataaaggcg aaacagcgat gaccatcaac ggcccgtggg catggtccaa catcgacacc
      841 agcaaagtga attatggtgt aacggtactg ccgaccttca agggtcaacc atccaaaccg
      901 ttcgttggcg tgctgagcgc aggtattaac gccgccagtc cgaacaaaga gctggcaaaa
      961 gagttcctcg aaaactatct gctgactgat gaaggtctgg aagcggttaa taaagacaaa
     1021 ccgctgggtg ccgtagcgct gaagtcttac gaggaagagt tggcgaaaga tccacgtatt
     1081 gccgccacta tggaaaacgc ccagaaaggt gaaatcatgc cgaacatccc gcagatgtcc
     1141 gctttctggt atgccgtgcg tactgcggtg atcaacgccg ccagcggtcg tcagactgtc
     1201 gatgaagccc tgaaagacgc gcagactaat tcgagctcga acaacaacaa caataacaat
     1261 aacaacaacc tcgggatcga ggaaaacctg tacttccaat ccaatgcaat gtctgaagct
     1321 cctaaaaagc gctggtacgt cgttcaggcg ttttccggtt ttgaaggccg cgtagcaacg
     1381 tcgctgcgtg agcatatcaa attacacaac atggaagatt tgtttggtga agtcatggta
     1441 ccaaccgaag aagtggttga aatccgtggc ggtcagcgtc gcaaaagcga acgtaaattc
     1501 ttccctggct acgtcctcgt tcagatggtg atgaacgacg cgagctggca cctggtgcgc
     1561 agcgtaccgc gtgtgatggg cttcatcggc ggtacttccg atcgtcctgc gccaatcagc
     1621 gataaagaag tcgatgcgat tatgaaccgc ctgcagcagg ttggttaata acattggaag
     1681 tggataacgg atccgaattc gagcgccgtc gacaagcttg cggccgcact cgagcaccac
     1741 caccaccacc actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     1801 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     1861 tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg
     1921 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     1981 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2041 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     2101 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2161 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2221 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     2281 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaac
     2341 tagtaacgtt tacaatttca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     2401 gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac
     2461 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     2521 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     2581 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     2641 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     2701 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
     2761 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     2821 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     2881 cgcaggaaca ctgccagcgc atcaacaatg ttttcacctg aatcaggata ttcttctaat
     2941 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     3001 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     3061 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     3121 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     3181 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa
     3241 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
     3301 agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3361 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3421 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3481 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     3541 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3601 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3661 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3721 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3781 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3841 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3901 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3961 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4021 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     4081 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     4141 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4201 ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     4261 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac
     4321 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     4381 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     4441 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
     4501 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
     4561 ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc
     4621 tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg
     4681 atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc
     4741 ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag
     4801 gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg
     4861 gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg
     4921 ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg
     4981 attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga
     5041 gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc ttctcgccga
     5101 aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata
     5161 ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga
     5221 cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg
     5281 cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca
     5341 agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc
     5401 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5461 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt
     5521 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg
     5581 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata
     5641 taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc
     5701 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc
     5761 atcgcagtgg gaacgatacc ctcattcagc atttgcatgg tttgttgaaa accggacatg
     5821 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta
     5881 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
     5941 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg
     6001 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
     6061 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
     6121 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
     6181 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
     6241 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
     6301 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
     6361 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg
     6421 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
     6481 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
     6541 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat
     6601 taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc
     6661 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
     6721 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     6781 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     6841 gtagaggatc gagatctcga tcccgcgaaa t
//
