LOCUS       sequence-247690-        7357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresion of RGG_GFP-RGG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7357)
  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 7357)
  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: pcDNA_RGG-GFP-RGG_withTEVsite
FEATURES             Location/Qualifiers
     source          1..7357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..29)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        201..580
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        581..784
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     735..755
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     829..848
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        829..847
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1423..1443
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(1444..1446,1447..1449,1450..2160)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1489..1510)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2914..2931)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2920..3144
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3190..3618
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3277..3296)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3487..3508
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3627..3647)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3632..3961
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3812..3947
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3874..3893
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4028..4822
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4082..4101)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4692..4711
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4996..5117
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5033..5052)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5087..5106
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5166..5182)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5166..5182)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5179..5201)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5190..5206
                     /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)."
     promoter        complement(join(5214..5220,5221..5238,5239..5244))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5259..5280
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5397..5414)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5568..6156)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5648..5667)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6327..7118,7119..7187))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6950..6969
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7188..7292)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
       61 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
      121 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
      181 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
      241 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      301 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      361 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      421 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      481 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      541 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      601 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      661 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      721 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      781 agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca
      841 ctatagggag acccaagctg gctagcgttt aaacttaagc ttggtaccga gctcggatcc
      901 atggaatcga atcaatctaa caacggtgga tcgggaaacg cagcattgaa tcgtgggggc
      961 cgttacgtac caccacactt gagaggcgga gatggtggtg ccgccgctgc tgcctcagcc
     1021 ggaggcgacg acagacgcgg tggggccggg ggtggtggct atcgtagagg tggtggaaat
     1081 tcagggggcg gcggcggagg aggctacgat cgtggataca acgacaaccg ggatgaccgt
     1141 gacaatcggg gcggctccgg cggttatggc cgggacagaa attacgagga tcgcggttat
     1201 aacgggggtg gtgggggagg aggcaatcgc ggctacaata ataaccgcgg tgggggtgga
     1261 ggcggctata atcggcagga ccgtggagac ggcggttcta gtaatttctc gcggggaggg
     1321 tataacaaca gagatgaggg ctcagacaac agaggctcag gcagatcata caacaacgat
     1381 cggcgcgaca atggtggtga cgggggcgga ggcggttccc tcgagaattt atatttccag
     1441 ggcatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1501 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1561 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     1621 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgatatcc agatcatatg
     1681 aagcagcacg atttctttaa aagtgcgatg cccgagggtt atgttcagga gcgcaccatc
     1741 ttcttcaagg acgacgggaa ttataagacc cgcgctgagg ttaagtttga aggggatacc
     1801 cttgttaaca gaattgagct gaagggtata gactttaaag aagatggcaa tatattgggg
     1861 cacaagctcg aatataatta caacagtcat aacgtatata tcatggctga caagcaaaag
     1921 aatggcataa aagtgaattt taagatacga cacaatatag aggatggtag cgtccagctt
     1981 gcggatcatt accagcagaa tacaccaatt ggtgacggcc cagtactgct gccggataat
     2041 cactatctca gtacccagag tgcattgtcc aaagacccga acgagaaacg cgatcacatg
     2101 gtactgctgg aatttgtaac ggctgctgga attacattgg gaatggacga gttgtacaag
     2161 gaattcggag gcggtggaag cgagtctaat cagagcaata acgggggtag tggcaatgcc
     2221 gcgctcaatc ggggaggccg ctacgtgcct cctcacctca gagggggcga tgggggcgcc
     2281 gctgccgctg cgagcgccgg tggcgatgat cggcggggag gagcgggtgg aggtggttat
     2341 cgccgaggtg gcgggaacag tgggggtgga ggaggcggtg gatatgatcg aggttacaac
     2401 gacaatcggg acgatcgaga taacagaggg ggttcaggtg gatacggaag agacaggaat
     2461 tacgaggaca gaggatacaa tggcggaggc ggtgggggag gtaatagagg atacaacaat
     2521 aatcgaggag gcgggggggg agggtacaat cgccaagacc gcggtgatgg aggaagttct
     2581 aactttagca gaggtggata caacaatagg gatgaaggct cagacaacag aggttctggt
     2641 cgaagctaca ataacgacag acgcgacaac ggtggcgatg ggtaataatc tagagggccc
     2701 gtttatcgag gattaccact acgctacgtg tacaaacgag gacctgatgc actttgacga
     2761 cttgggatcg acgagagcag cgcgacaacg gtggcgatgg gtaataatct agagggcccg
     2821 tttgtcgagg attaccacta cgctacgtgc ctcctcacct cagagggggc gatgggtaat
     2881 aatctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     2941 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     3001 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     3061 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     3121 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     3181 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3241 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3301 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     3361 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3421 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3481 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3541 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     3601 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3661 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3721 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3781 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3841 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3901 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3961 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     4021 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4081 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4141 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4201 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4261 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4321 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4381 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4441 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4501 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     4561 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4621 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4681 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4741 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4801 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4861 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4921 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4981 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     5041 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     5101 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     5161 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     5221 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     5281 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5341 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5401 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5461 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5521 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5581 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5641 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5701 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5761 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5821 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5881 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5941 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6001 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6061 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6121 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6181 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6241 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6301 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6361 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6421 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6481 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6541 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6601 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6661 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6721 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6781 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6841 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6901 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6961 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7021 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7081 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7141 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7201 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7261 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7321 gtcgacggat cgggagatct cccgatcccc tatggtg
//
