LOCUS       sequence-245504-        5237 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of RGG-RGG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5237)
  AUTHORS   Schuster BS, Reed EH, Parthasarathy R, Jahnke CN, Caldwell RM, 
            Bermudez JG, Ramage H, Good MC, Hammer DA
  TITLE     Controllable protein phase separation and modular recruitment to 
            form responsive membraneless organelles.
  JOURNAL   Nat Commun. 2018 Jul 30;9(1):2985. doi: 10.1038/s41467-018-05403-1.
  PUBMED    30061688
REFERENCE   2  (bases 1 to 5237)
  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: "Nat 
            Commun."; date: "2018-07-30"; volume: "9(1):2985. doi"; pages: " 
            10.1038/s41467-018-05403-1"
COMMENT     Sequence Label: pET_RGG-RGG_withTEVsite
FEATURES             Location/Qualifiers
     source          1..5237
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     86..105
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        86..104
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    105..129
                     /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             144..166
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             684..704
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1218..1235
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1288..1306)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1302..1349
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1386..1841
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1473..1492)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1683..1704
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1956..2771)
                     /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(1956..2771)
                     /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     2679..2698
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2763..2782
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2924..3512
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3413..3432
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3666..3683
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(3854..4936)
                     /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     4898..4917
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4937..5014)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(5220..5237,1..2)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
       61 gatcgagatc tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat
      121 aacaattccc ctctagaaat aattttgttt aactttaaga aggagatata catatggaat
      181 ccaatcaatc caataacggt ggctcgggta atgccgctct gaatcgtggt ggccgctacg
      241 tgccgccgca tctgcgtggt ggcgatggtg gtgcagcagc agctgcatct gctggcggtg
      301 atgaccgtcg cggcggtgcg ggcggtggcg gttatcgtcg cggcggtggc aatagtggtg
      361 gcggtggcgg tggcggttat gatcgtggtt acaatgacaa ccgtgatgac cgcgataacc
      421 gtggcggttc cggcggttac ggtcgtgatc gtaattatga agaccgtggt tacaacggcg
      481 gtggcggtgg cggtggcaat cgtggctata acaataaccg tggtggcggt ggcggtggct
      541 acaatcgcca ggatcgtggt gacggtggca gctctaactt ttcgcgcggt ggctataata
      601 accgtgatga aggcagcgac aaccgcggta gtggccgttc ctacaataac gatcgtcgcg
      661 acaatggtgg cgatggcgaa ttcgaaaatt tgtatttcca aggcggatcc gaatctaatc
      721 aatcgaataa cggcgggagc ggtaacgcag cgcttaacag aggggggaga tacgtccccc
      781 ctcatttgag agggggtgat gggggagcgg cagcagctgc ttccgctgga ggtgatgacc
      841 gtcggggtgg agctgggggc ggcggctatc gcagaggggg cggtaatagc ggagggggcg
      901 ggggtggcgg gtatgacaga gggtacaacg ataaccggga tgaccgggat aatcgtgggg
      961 ggtccggcgg ctatggccgc gacagaaatt acgaagatag aggatacaat ggtggcggcg
     1021 gtggcggcgg aaatcggggt tacaataata atagaggcgg gggaggtggg ggttacaacc
     1081 gtcaggatcg gggtgacgga gggtcatcga atttttccag aggaggctat aacaatcggg
     1141 atgagggtag tgataacaga ggctctggcc gttcttataa caatgatcgc agagataatg
     1201 ggggcgatgg actcgagcac caccaccacc accactgaga tccggctgct aacaaagccc
     1261 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
     1321 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga
     1381 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     1441 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     1501 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     1561 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     1621 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     1681 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     1741 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     1801 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcggggt
     1861 acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1921 tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc gagcatcaaa
     1981 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     2041 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     2101 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     2161 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt
     2221 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     2281 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     2341 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     2401 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     2461 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     2521 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     2581 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     2641 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     2701 tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga
     2761 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     2821 gaccaaaatc ccttaacgtg acctaggtca tgaccaaaat cccttaacgt gagttttcgt
     2881 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2941 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3001 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3061 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3121 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3181 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3241 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3301 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3361 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3421 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3481 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3541 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3601 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3661 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcca taatccatga taaagaagac
     3721 agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
     3781 gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt
     3841 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     3901 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
     3961 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
     4021 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
     4081 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg
     4141 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     4201 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     4261 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     4321 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     4381 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     4441 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     4501 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     4561 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     4621 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     4681 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     4741 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     4801 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     4861 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     4921 acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca
     4981 taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc
     5041 gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca
     5101 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     5161 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
     5221 gtgatgtcgg cgatata
//
