LOCUS       sequence-196622-        8174 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses iPOLYMER component in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8174)
  AUTHORS   Nakamura H, Lee AA, Afshar AS, Watanabe S, Rho E, Razavi S, Suarez 
            A, Lin YC, Tanigawa M, Huang B, DeRose R, Bobb D, Hong W, Gabelli 
            SB, Goutsias J, Inoue T
  TITLE     Intracellular production of hydrogels and synthetic RNA granules by 
            multivalent molecular interactions.
  JOURNAL   Nat Mater. 2017 Nov 6. pii: nmat5006. doi: 10.1038/nmat5006.
  PUBMED    29115293
REFERENCE   2  (bases 1 to 8174)
  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 Mater. 
            2017 Nov 6. pii: nmat5006. doi: 10.1038/nmat5006."
COMMENT     Sequence Label: TIA1 RRM-mCherry-iLIDx6
FEATURES             Location/Qualifiers
     source          1..8174
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      597..606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1266..1275
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1272..1979
                     /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(1417..1435)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1685..1704
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4962..5083
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4999..5018)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5053..5072
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5090..5545)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5227..5248)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5439..5458
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5572..5676
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5678..6035
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5678..5869
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5701..5721)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5886..6021
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6070..6864
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6070..6864
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6124..6143)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6734..6753
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7052..7071)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7096..7143
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7472..8060
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7961..7980
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcca
      601 ccatggagga cgagatgccc aagactctat acgtcggtaa cctttccaga gatgtgacag
      661 aagctctaat tctgcaactc tttagccaga ttggaccttg taaaaactgc aaaatgatta
      721 tggatacagc tggaaatgat ccctattgtt ttgtggagtt tcatgagcat cgtcatgcag
      781 ctgcagcatt agctgctatg aatggacgga agataatggg taaggaagtc aaagtgaatt
      841 gggcaacaac ccctagcagt caaaagaaag atacaagcaa tcatttccat gtctttgttg
      901 gtgatctcag cccagaaatt acaactgaag atataaaagc tgcttttgca ccatttggaa
      961 gaatatcaga tgcccgagtg gtaaaagaca tggcaacagg aaagtctaag ggatatggct
     1021 ttgtctcctt tttcaacaaa tgggatgctg aaaacgccat tcaacagatg ggtggccagt
     1081 ggcttggtgg aagacaaatc agaactaact gggcaacccg aaagcctccc gctccaaaga
     1141 gtacatatga gtcaaatacc aaacagctat catatgatga ggttgtaaat cagtctagtc
     1201 caagcaactg tactgtatac tgtggaggtg ttacttctgg gctaacagaa caactaggac
     1261 cggtcgccac catggtaagc aagggcgagg aggataacat ggccatcatc aaggagttca
     1321 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
     1381 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
     1441 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
     1501 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
     1561 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
     1621 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
     1681 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc
     1741 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
     1801 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     1861 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
     1921 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt
     1981 ccggactcag atctagcgct ggaggcagcg ctggcggaga gttcctggca accacactgg
     2041 agcggatcga gaagaatttt gtgatcaccg acccccgcct gcctgataac ccaatcatct
     2101 tcgccagcga ctcctttctg cagctgacag agtacagcag ggaggagatc ctgggcagga
     2161 attgcagatt cctgcagggc cccgagacag atagagccac agtgcggaag atccgcgacg
     2221 ccatcgataa ccagaccgag gtgacagtgc agctgatcaa ttataccaag tccggcaaga
     2281 agttctggaa cgtgtttcac ctgcagccta tgcgggacta caagggcgat gtgcagtact
     2341 tcatcggagt gcagctggac ggaaccgaga ggctgcacgg agcagccgag agagaggccg
     2401 tgtgcctgat caagaagaca gccttccaga tcgctgaggc cgccaacgat gagaactact
     2461 ttaagctttc cgcaggcggg tccgcagggg gcgagttcct ggccaccaca ctggagagaa
     2521 ttgagaagaa tttcgtcatc accgacccca gactgccaga caatcctatc atcttcgcct
     2581 ctgacagctt tctgcagctg accgaatact ctagagaaga gatcctgggc aggaactgta
     2641 gattcctgca gggaccagag acagataggg caacagtgcg gaagatcagg gacgcaatcg
     2701 ataatcagac tgaagtcacc gtgcagctga tcaactatac caagagtggc aagaagtttt
     2761 ggaacgtgtt ccacctgcag cccatgcgcg attataaagg cgatgtgcaa tacttcatcg
     2821 gcgtgcagct ggacggcacc gagcggctgc acggagcagc agagagggaa gccgtgtgcc
     2881 tgatcaagaa gacagcgttt cagattgccg aagctgctaa tgatgaaaat tacttcgtcg
     2941 actctgccgg gggatctgcc ggaggcgagt tcctggctac cacactggaa agaattgaga
     3001 agaattttgt cattaccgac cccagactgc ccgacaatcc catcatcttc gcctccgact
     3061 cttttctgca gctgaccgag tattctagag aggagatcct gggaagaaac tgcagattcc
     3121 tgcagggacc tgagacagac agggctacag tgcggaaaat tagagatgct attgacaatc
     3181 agactgaagt cactgtccag ctgatcaatt acactaagtc tggcaagaag ttctggaacg
     3241 tgtttcacct gcaacccatg agagattata agggcgatgt gcagtatttc atcggggtcc
     3301 agctggatgg cactgagagg ctgcatggcg ccgctgaaag ggaggccgtg tgcctgatca
     3361 agaagacagc ttttcagatt gctgaagctg ccaacgacga gaattatttt ggatctagcg
     3421 ctggaggcag cgctggcgga gagttcctgg caaccacact ggagcggatc gagaagaatt
     3481 ttgtgatcac cgacccccgc ctgcctgata acccaatcat cttcgccagc gactcctttc
     3541 tgcagctgac agagtacagc agggaggaga tcctgggcag gaattgcaga ttcctgcagg
     3601 gccccgagac agatagagcc acagtgcgga agatccgcga cgccatcgat aaccagaccg
     3661 aggtgacagt gcagctgatc aattatacca agtccggcaa gaagttctgg aacgtgtttc
     3721 acctgcagcc tatgcgggac tacaagggcg atgtgcagta cttcatcgga gtgcagctgg
     3781 acggaaccga gaggctgcac ggagcagccg agagagaggc cgtgtgcctg atcaagaaga
     3841 cagccttcca gatcgctgag gccgccaacg atgagaacta ctttaagctt tccgcaggcg
     3901 ggtccgcagg gggcgagttc ctggccacca cactggagag aattgagaag aatttcgtca
     3961 tcaccgaccc cagactgcca gacaatccta tcatcttcgc ctctgacagc tttctgcagc
     4021 tgaccgaata ctctagagaa gagatcctgg gcaggaactg tagattcctg cagggaccag
     4081 agacagatag ggcaacagtg cggaagatca gggacgcaat cgataatcag actgaagtca
     4141 ccgtgcagct gatcaactat accaagagtg gcaagaagtt ttggaacgtg ttccacctgc
     4201 agcccatgcg cgattataaa ggcgatgtgc aatacttcat cggcgtgcag ctggacggca
     4261 ccgagcggct gcacggagca gcagagaggg aagccgtgtg cctgatcaag aagacagcgt
     4321 ttcagattgc cgaagctgct aatgatgaaa attacttcgt cgactctgcc gggggatctg
     4381 ccggaggcga gttcctggct accacactgg aaagaattga gaagaatttt gtcattaccg
     4441 accccagact gcccgacaat cccatcatct tcgcctccga ctcttttctg cagctgaccg
     4501 agtattctag agaggagatc ctgggaagaa actgcagatt cctgcaggga cctgagacag
     4561 acagggctac agtgcggaaa attagagatg ctattgacaa tcagactgaa gtcactgtcc
     4621 agctgatcaa ttacactaag tctggcaaga agttctggaa cgtgtttcac ctgcaaccca
     4681 tgagagatta taagggcgat gtgcagtatt tcatcggggt ccagctggat ggcactgaga
     4741 ggctgcatgg cgccgctgaa agggaggccg tgtgcctgat caagaagaca gcttttcaga
     4801 ttgctgaagc tgccaacgac gagaattatt ttggatccac cggatctaga taactgatca
     4861 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     4921 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     4981 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5041 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc
     5101 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     5161 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     5221 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     5281 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     5341 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     5401 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     5461 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     5521 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5581 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5641 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     5701 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5761 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     5821 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     5881 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     5941 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     6001 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     6061 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     6121 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     6181 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     6241 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     6301 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     6361 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     6421 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     6481 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     6541 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     6601 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     6661 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     6721 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     6781 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     6841 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     6901 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     6961 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     7021 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     7081 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     7141 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     7201 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     7261 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     7321 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     7381 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     7441 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     7501 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     7561 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     7621 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     7681 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     7741 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7801 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7861 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7921 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     7981 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     8041 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     8101 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     8161 ttaccgccat gcat
//
