LOCUS       sequence-245541-        5222 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresion of RGG-RGG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5222)
  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 5222)
  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
FEATURES             Location/Qualifiers
     source          1..5222
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     30..49
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     133..152
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        133..151
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    152..176
                     /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             191..213
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1250..1267
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1320..1338)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1334..1381
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1418..1873
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1505..1524)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1715..1736
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1988..2803)
                     /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(1988..2803)
                     /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     2711..2730
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2795..2814
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2956..3544
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3445..3464
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3698..3715
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(3886..4968)
                     /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     4930..4949
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4969..5046)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
       61 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg
      121 atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac aattcccctc
      181 tagaaataat tttgtttaac tttaagaagg agatatacat atggaatcca atcaatccaa
      241 taacggtggc tcgggtaatg ccgctctgaa tcgtggtggc cgctacgtgc cgccgcatct
      301 gcgtggtggc gatggtggtg cagcagcagc tgcatctgct ggcggtgatg accgtcgcgg
      361 cggtgcgggc ggtggcggtt atcgtcgcgg cggtggcaat agtggtggcg gtggcggtgg
      421 cggttatgat cgtggttaca atgacaaccg tgatgaccgc gataaccgtg gcggttccgg
      481 cggttacggt cgtgatcgta attatgaaga ccgtggttac aacggcggtg gcggtggcgg
      541 tggcaatcgt ggctataaca ataaccgtgg tggcggtggc ggtggctaca atcgccagga
      601 tcgtggtgac ggtggcagct ctaacttttc gcgcggtggc tataataacc gtgatgaagg
      661 cagcgacaac cgcggtagtg gccgttccta caataacgat cgtcgcgaca atggtggcga
      721 tggcgaattc gggaagctta tggaatccaa tcaatccaat aacggtggct cgggtaatgc
      781 cgctctgaat cgtggtggcc gctacgtgcc gccgcatctg cgtggtggcg atggtggtgc
      841 agcagcagct gcatctgctg gcggtgatga ccgtcgcggc ggtgcgggcg gtggcggtta
      901 tcgtcgcggc ggtggcaata gtggtggcgg tggcggtggc ggttatgatc gtggttacaa
      961 tgacaaccgt gatgaccgcg ataaccgtgg cggttccggc ggttacggtc gtgatcgtaa
     1021 ttatgaagac cgtggttaca acggcggtgg cggtggcggt ggcaatcgtg gctataacaa
     1081 taaccgtggt ggcggtggcg gtggctacaa tcgccaggat cgtggtgacg gtggcagctc
     1141 taacttttcg cgcggtggct ataataaccg tgatgaaggc agcgacaacc gcggtagtgg
     1201 ccgttcctac aataacgatc gtcgcgacaa tggtggcgat ggcctcgagc accaccacca
     1261 ccaccactga gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     1321 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     1381 gctgaaagga ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta
     1441 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     1501 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     1561 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     1621 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     1681 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     1741 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     1801 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     1861 acgcttacaa tttaggtggc acttttcggg gtacccaggt ggcacttttc ggggaaatgt
     1921 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgaa
     1981 ttaattctta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
     2041 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
     2101 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
     2161 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
     2221 tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa
     2281 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     2341 gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
     2401 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     2461 caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc
     2521 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     2581 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     2641 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
     2701 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     2761 atcgcggcct agagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac
     2821 tgtttatgta agcagacagt tttattgttc atgaccaaaa tcccttaacg tgacctaggt
     2881 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     2941 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3001 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3061 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3121 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3181 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3241 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3301 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3361 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3421 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3481 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3541 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3601 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     3661 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     3721 ggaagagcgc cataatccat gataaagaag acagtcataa gtgcggcgac gatagtcatg
     3781 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     3841 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     3901 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3961 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     4021 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     4081 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     4141 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     4201 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     4261 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     4321 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     4381 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     4441 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     4501 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     4561 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     4621 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     4681 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     4741 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     4801 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     4861 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     4921 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     4981 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     5041 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
     5101 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     5161 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     5221 gc
//
