LOCUS       sequence-223538-        6498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses E. coli Rho.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6498)
  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 6498)
  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: EcRho
FEATURES             Location/Qualifiers
     source          1..6498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1282..1299
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1352..1370)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1366..1413
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1450..1905
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1537..1556)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1747..1768
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1997..2812)
                     /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(1997..2812)
                     /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     2720..2739
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2804..2823
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2934..3522
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3423..3442
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3676..3693
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3708..3847
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3773..3792)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3933..3955
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3949..4140)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4949..6031)
                     /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     5993..6012
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6032..6109)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6315..6334
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6418..6437
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6418..6436
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6437..6461
                     /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             6476..6498
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 tataccatgg gccatatgaa tcttaccgaa ttaaagaata cgccggtttc tgaactgatc
       61 actctcggcg aaaatatggg gctggaaaac ctggctcgta tgcgtaagca ggacattatt
      121 tttgccatcc tgaagcagca cgcaaagagt ggcgaagata tctttggtga tggcgtactg
      181 gagatattgc aggatggatt tggtttcctc cgttccgcag acagctccta cctcgccggt
      241 cctgatgaca tctacgtttc ccctagccaa atccgccgtt tcaacctccg cactggtgat
      301 accatctctg gtaagattcg cccaccgaaa gaaggtgaac gctattttgc gctgctgaaa
      361 gttaacgaag ttaacttcga caaacctgaa aacgcccgca acaaaatcct ctttgagaac
      421 ttaaccccgc tgcacgcaaa ctctcgtctg cgtatggaac gtggtaacgg ttctactgaa
      481 gatttaactg ctcgcgtact ggatctggca tcacctatcg gtcgtggtca gcgtggtctg
      541 attgtggcac cgccgaaagc cggtaaaacc atgctgctgc agaacattgc tcagagcatt
      601 gcttacaacc acccggattg tgtgctgatg gttctgctga tcgacgaacg tccggaagaa
      661 gtaaccgaga tgcagcgtct ggtaaaaggt gaagttgttg cttctacctt tgacgaaccc
      721 gcatctcgcc acgttcaggt tgcggaaatg gtgatcgaga aggccaaacg cctggttgag
      781 cacaagaaag acgttatcat tctgctcgac tccatcactc gtctggcgcg cgcttacaac
      841 accgttgttc cggcgtcagg taaagtgttg accggtggtg tggatgccaa cgccctgcat
      901 cgtccgaaac gcttctttgg tgcggcgcgt aacgtggaag agggcggcag cctgaccatt
      961 atcgcgacgg cgcttatcga taccggttct aaaatggacg aagttatcta cgaagagttt
     1021 aaaggtacag gcaacatgga actgcacctc tctcgtaaga tcgctgaaaa acgcgtcttc
     1081 ccggctatcg actacaaccg ttctggtacc cgtaaagaag agctgctcac gactcaggaa
     1141 gaactgcaga aaatgtggat cctgcgcaaa atcattcacc cgatgggcga aatcgatgca
     1201 atggaattcc tcattaataa actggcaatg accaagacca atgacgattt cttcgaaatg
     1261 atgaaacgct cataactcga gcaccaccac caccaccact gagatccggc tgctaacaaa
     1321 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
     1381 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggattg
     1441 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     1501 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     1561 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     1621 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     1681 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     1741 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     1801 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     1861 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     1921 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1981 gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca
     2041 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     2101 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     2161 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     2221 caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga
     2281 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     2341 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat
     2401 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     2461 cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg
     2521 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     2581 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     2641 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg
     2701 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     2761 tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc ataacacccc
     2821 ttgtattact gtttatgtaa gcagacagtt ttattgttca tgaccaaaat cccttaacgt
     2881 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2941 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3001 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3061 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     3121 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3181 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3241 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3301 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3361 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3421 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3481 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3541 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3601 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3661 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     3721 tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct
     3781 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     3841 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     3901 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     3961 accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag
     4021 cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt
     4081 taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac
     4141 tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga
     4201 gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga
     4261 gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg
     4321 ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat
     4381 gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac
     4441 acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc
     4501 gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca
     4561 gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtgggg ccgccatgcc
     4621 ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg
     4681 agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca
     4741 gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg
     4801 catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa
     4861 ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt
     4921 gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4981 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5041 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
     5101 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
     5161 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca
     5221 ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
     5281 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
     5341 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
     5401 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
     5461 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
     5521 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag
     5581 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
     5641 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
     5701 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
     5761 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
     5821 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
     5881 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
     5941 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
     6001 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
     6061 ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga
     6121 cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg
     6181 agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
     6241 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     6301 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     6361 gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa
     6421 tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa ataattttgt
     6481 ttaactttaa gaaggaga
//
