LOCUS       sequence-245479-        4703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of RGG domain from LAF-1 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4703)
  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 4703)
  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
FEATURES             Location/Qualifiers
     source          1..4703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..62
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     146..165
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        146..164
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    165..189
                     /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             204..226
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             744..761
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(814..832)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      828..875
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      912..1367
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(999..1018)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1209..1230
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1482..2297)
                     /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(1482..2297)
                     /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     2205..2224
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2289..2308
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2450..3038
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2939..2958
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3192..3209
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(3380..4462)
                     /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     4424..4443
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4463..4540)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
       61 ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag
      121 gatcgagatc tcgatcccgc gaaattaata cgactcacta taggggaatt gtgagcggat
      181 aacaattccc ctctagaaat aattttgttt aactttaaga aggagatata catatggaat
      241 ccaatcaatc caataacggt ggctcgggta atgccgctct gaatcgtggt ggccgctacg
      301 tgccgccgca tctgcgtggt ggcgatggtg gtgcagcagc agctgcatct gctggcggtg
      361 atgaccgtcg cggcggtgcg ggcggtggcg gttatcgtcg cggcggtggc aatagtggtg
      421 gcggtggcgg tggcggttat gatcgtggtt acaatgacaa ccgtgatgac cgcgataacc
      481 gtggcggttc cggcggttac ggtcgtgatc gtaattatga agaccgtggt tacaacggcg
      541 gtggcggtgg cggtggcaat cgtggctata acaataaccg tggtggcggt ggcggtggct
      601 acaatcgcca ggatcgtggt gacggtggca gctctaactt ttcgcgcggt ggctataata
      661 accgtgatga aggcagcgac aaccgcggta gtggccgttc ctacaataac gatcgtcgcg
      721 acaatggtgg cgatggcctc gagcaccacc accaccacca ctgagatccg gctgctaaca
      781 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
      841 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat
      901 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
      961 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     1021 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     1081 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     1141 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     1201 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     1261 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     1321 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     1381 cggggtaccc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1441 tctaaataca ttcaaatatg tatccgctca tgaattaatt cttagaaaaa ctcatcgagc
     1501 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     1561 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     1621 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     1681 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     1741 aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     1801 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
     1861 acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
     1921 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
     1981 gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
     2041 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
     2101 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
     2161 ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     2221 tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca agacgtttcc
     2281 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt
     2341 gttcatgacc aaaatccctt aacgtgacct aggtcatgac caaaatccct taacgtgagt
     2401 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2461 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2521 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2581 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2641 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2701 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2761 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2821 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2881 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2941 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3001 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3061 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3121 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     3181 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgccataat ccatgataaa
     3241 gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac
     3301 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     3361 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3421 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     3481 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     3541 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     3601 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     3661 tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     3721 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     3781 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     3841 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
     3901 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
     3961 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
     4021 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
     4081 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
     4141 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
     4201 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
     4261 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
     4321 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
     4381 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
     4441 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
     4501 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
     4561 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc
     4621 gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct
     4681 gccaccatac ccacgccgaa aca
//
