LOCUS       sequence-245487-        7330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of RGG-RFP-RGG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7330)
  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 7330)
  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-RFP-RGG
FEATURES             Location/Qualifiers
     source          1..7330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        120..499
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        500..703
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     654..674
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     748..767
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        748..766
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1345..2052
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1490..1508)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1758..1777
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2806..2823)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2812..3036
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3082..3510
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3169..3188)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3379..3400
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3519..3539)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3524..3853
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3704..3839
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3766..3785
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3920..4714
                     /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(3974..3993)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4584..4603
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4888..5009
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4925..4944)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4979..4998
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5058..5074)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5058..5074)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5071..5093)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5082..5098
                     /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(5106..5112,5113..5130,5131..5136))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5151..5172
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5289..5306)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5460..6048)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5540..5559)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6219..7010,7011..7079))
                     /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     6842..6861
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7080..7184)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7259..7278)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
       61 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
      121 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
      181 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      241 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      301 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      361 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      421 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      481 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      541 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      601 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      661 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga
      721 acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg
      781 ctagcgttta aacttaagct tggtaccgag ctcggatcca tggaatcgaa tcaatctaac
      841 aacggtggat cgggaaacgc agcattgaat cgtgggggcc gttacgtacc accacacttg
      901 agaggcggag atggtggtgc cgccgctgct gcctcagccg gaggcgacga cagacgcggt
      961 ggggccgggg gtggtggcta tcgtagaggt ggtggaaatt cagggggcgg cggcggagga
     1021 ggctacgatc gtggatacaa cgacaaccgg gatgaccgtg acaatcgggg cggctccggc
     1081 ggttatggcc gggacagaaa ttacgaggat cgcggttata acgggggtgg tgggggagga
     1141 ggcaatcgcg gctacaataa taaccgcggt gggggtggag gcggctataa tcggcaggac
     1201 cgtggagacg gcggttctag taatttctcg cggggagggt ataacaacag agatgagggc
     1261 tcagacaaca gaggctcagg cagatcatac aacaacgatc ggcgcgacaa tggtggtgac
     1321 gggggcggag gcggttccct cgagatggtg agcaagggcg aggaggataa catggccatc
     1381 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc
     1441 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     1501 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac
     1561 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc
     1621 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg
     1681 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc
     1741 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc
     1801 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg
     1861 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg
     1921 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac
     1981 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac
     2041 gagctgtaca aggaattcgg aggcggtgga agcgagtcta atcagagcaa taacgggggt
     2101 agtggcaatg ccgcgctcaa tcggggaggc cgctacgtgc ctcctcacct cagagggggc
     2161 gatgggggcg ccgctgccgc tgcgagcgcc ggtggcgatg atcggcgggg aggagcgggt
     2221 ggaggtggtt atcgccgagg tggcgggaac agtgggggtg gaggaggcgg tggatatgat
     2281 cgaggttaca acgacaatcg ggacgatcga gataacagag ggggttcagg tggatacgga
     2341 agagacagga attacgagga cagaggatac aatggcggag gcggtggggg aggtaataga
     2401 ggatacaaca ataatcgagg aggcgggggg ggagggtaca atcgccaaga ccgcggtgat
     2461 ggaggaagtt ctaactttag cagaggtgga tacaacaata gggatgaagg ctcagacaac
     2521 agaggttctg gtcgaagcta caataacgac agacgcgaca acggtggcga tgggtaataa
     2581 tctagagggc ccgtttatcg aggattacca ctacgctacg tgtacaaacg aggacctgat
     2641 gcactttgac gacttgggat cgacgagagc agcgcgacaa cggtggcgat gggtaataat
     2701 ctagagggcc cgtttgtcga ggattaccac tacgctacgt gcctcctcac ctcagagggg
     2761 gcgatgggta ataatctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt
     2821 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     2881 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     2941 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     3001 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     3061 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3121 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3181 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3241 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3301 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3361 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3421 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3481 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     3541 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3601 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3661 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3721 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3781 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3841 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     3901 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3961 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     4021 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     4081 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     4141 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     4201 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     4261 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     4321 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     4381 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     4441 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     4501 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     4561 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     4621 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     4681 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     4741 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4801 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4861 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     4921 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4981 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     5041 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5101 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5161 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5221 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5281 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5341 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5401 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5461 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5521 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5581 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5641 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5701 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5761 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5821 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5881 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5941 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6001 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6061 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6121 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6181 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6241 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6301 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6361 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6421 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6481 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6541 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6601 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6661 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6721 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6781 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6841 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6901 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6961 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7021 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7081 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7141 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7201 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta
     7261 caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg
     7321 tcgctgagta
//
