LOCUS       sequence-245489-        6260 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of RFP-SZ2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6260)
  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 6260)
  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_RFP-SZ2
FEATURES             Location/Qualifiers
     source          1..6260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        101..480
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        481..684
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     635..655
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     729..748
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        729..747
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             801..1508
                     /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(946..964)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1214..1233
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1512..1532
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1717..1734)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1723..1947
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1993..2421
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2080..2099)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2290..2311
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2430..2450)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2435..2764
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2615..2750
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2677..2696
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2831..3625
                     /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(2885..2904)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3495..3514
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3799..3920
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3836..3855)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3890..3909
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3969..3985)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3969..3985)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3982..4004)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3993..4009
                     /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(4017..4023,4024..4041,4042..4047))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4062..4083
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4200..4217)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4371..4959)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4451..4470)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5130..5921,5922..5990))
                     /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     5753..5772
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5991..6095)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6170..6189)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
       61 tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt
      121 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      181 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      241 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      301 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      361 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      421 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      481 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      541 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      601 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      661 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
      721 atcgaaatta atacgactca ctatagggag acccaagctg gctagcgttt aaacttaagc
      781 ttggtaccga gctcggatcc atggtgagca agggcgagga ggataacatg gccatcatca
      841 aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga
      901 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga
      961 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct
     1021 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg
     1081 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc
     1141 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact
     1201 tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc
     1261 ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg
     1321 acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc
     1381 tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca
     1441 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc
     1501 tgtacaaggg ggaaaatctg tatttccagg gagcgcgtaa cgcatatctt cgtaaaaaga
     1561 ttgcacgcct gaagaaggac aatttacaat tggaacgcga cgaacaaaat ttagagaaaa
     1621 tcattgctaa tctgcgcgac gagattgctc gcttagaaaa cgaagtcgca tcacatgagc
     1681 agtaatctag agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     1741 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     1801 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     1861 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     1921 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     1981 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     2041 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     2101 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     2161 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     2221 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     2281 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2341 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2401 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     2461 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     2521 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2581 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     2641 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     2701 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     2761 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     2821 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     2881 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     2941 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3001 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     3061 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     3121 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     3181 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     3241 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     3301 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     3361 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     3421 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     3481 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     3541 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     3601 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     3661 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     3721 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     3781 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3841 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3901 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     3961 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4021 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4081 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4141 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4201 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4261 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4321 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4381 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4441 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4501 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4561 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4621 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4681 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4741 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4801 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4861 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4921 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4981 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5041 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5101 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5161 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5221 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5281 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5341 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5401 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5461 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5521 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5581 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5641 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5701 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5761 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5821 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5881 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5941 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6001 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6061 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6121 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     6181 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     6241 agtgcgcgag caaaatttaa
//
