LOCUS       sequence-196613-        7515 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 7515)
  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 7515)
  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: mCherry-iLIDx6
FEATURES             Location/Qualifiers
     source          1..7515
                     /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      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1320
                     /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(758..776)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1026..1045
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4303..4424
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4340..4359)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4394..4413
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4431..4886)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4568..4589)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4780..4799
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4913..5017
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5019..5376
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5019..5210
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5042..5062)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5227..5362
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5411..6205
                     /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             5411..6205
                     /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(5465..5484)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6075..6094
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6393..6412)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6437..6484
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6813..7401
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7302..7321
                     /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 gctagcgcta
      601 ccggtcgcca ccatggtaag caagggcgag gaggataaca tggccatcat caaggagttc
      661 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      721 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
      781 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
      841 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
      901 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
      961 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     1021 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     1081 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     1141 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     1201 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     1261 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     1321 tccggactca gatctagcgc tggaggcagc gctggcggag agttcctggc aaccacactg
     1381 gagcggatcg agaagaattt tgtgatcacc gacccccgcc tgcctgataa cccaatcatc
     1441 ttcgccagcg actcctttct gcagctgaca gagtacagca gggaggagat cctgggcagg
     1501 aattgcagat tcctgcaggg ccccgagaca gatagagcca cagtgcggaa gatccgcgac
     1561 gccatcgata accagaccga ggtgacagtg cagctgatca attataccaa gtccggcaag
     1621 aagttctgga acgtgtttca cctgcagcct atgcgggact acaagggcga tgtgcagtac
     1681 ttcatcggag tgcagctgga cggaaccgag aggctgcacg gagcagccga gagagaggcc
     1741 gtgtgcctga tcaagaagac agccttccag atcgctgagg ccgccaacga tgagaactac
     1801 tttaagcttt ccgcaggcgg gtccgcaggg ggcgagttcc tggccaccac actggagaga
     1861 attgagaaga atttcgtcat caccgacccc agactgccag acaatcctat catcttcgcc
     1921 tctgacagct ttctgcagct gaccgaatac tctagagaag agatcctggg caggaactgt
     1981 agattcctgc agggaccaga gacagatagg gcaacagtgc ggaagatcag ggacgcaatc
     2041 gataatcaga ctgaagtcac cgtgcagctg atcaactata ccaagagtgg caagaagttt
     2101 tggaacgtgt tccacctgca gcccatgcgc gattataaag gcgatgtgca atacttcatc
     2161 ggcgtgcagc tggacggcac cgagcggctg cacggagcag cagagaggga agccgtgtgc
     2221 ctgatcaaga agacagcgtt tcagattgcc gaagctgcta atgatgaaaa ttacttcgtc
     2281 gactctgccg ggggatctgc cggaggcgag ttcctggcta ccacactgga aagaattgag
     2341 aagaattttg tcattaccga ccccagactg cccgacaatc ccatcatctt cgcctccgac
     2401 tcttttctgc agctgaccga gtattctaga gaggagatcc tgggaagaaa ctgcagattc
     2461 ctgcagggac ctgagacaga cagggctaca gtgcggaaaa ttagagatgc tattgacaat
     2521 cagactgaag tcactgtcca gctgatcaat tacactaagt ctggcaagaa gttctggaac
     2581 gtgtttcacc tgcaacccat gagagattat aagggcgatg tgcagtattt catcggggtc
     2641 cagctggatg gcactgagag gctgcatggc gccgctgaaa gggaggccgt gtgcctgatc
     2701 aagaagacag cttttcagat tgctgaagct gccaacgacg agaattattt tggatctagc
     2761 gctggaggca gcgctggcgg agagttcctg gcaaccacac tggagcggat cgagaagaat
     2821 tttgtgatca ccgacccccg cctgcctgat aacccaatca tcttcgccag cgactccttt
     2881 ctgcagctga cagagtacag cagggaggag atcctgggca ggaattgcag attcctgcag
     2941 ggccccgaga cagatagagc cacagtgcgg aagatccgcg acgccatcga taaccagacc
     3001 gaggtgacag tgcagctgat caattatacc aagtccggca agaagttctg gaacgtgttt
     3061 cacctgcagc ctatgcggga ctacaagggc gatgtgcagt acttcatcgg agtgcagctg
     3121 gacggaaccg agaggctgca cggagcagcc gagagagagg ccgtgtgcct gatcaagaag
     3181 acagccttcc agatcgctga ggccgccaac gatgagaact actttaagct ttccgcaggc
     3241 gggtccgcag ggggcgagtt cctggccacc acactggaga gaattgagaa gaatttcgtc
     3301 atcaccgacc ccagactgcc agacaatcct atcatcttcg cctctgacag ctttctgcag
     3361 ctgaccgaat actctagaga agagatcctg ggcaggaact gtagattcct gcagggacca
     3421 gagacagata gggcaacagt gcggaagatc agggacgcaa tcgataatca gactgaagtc
     3481 accgtgcagc tgatcaacta taccaagagt ggcaagaagt tttggaacgt gttccacctg
     3541 cagcccatgc gcgattataa aggcgatgtg caatacttca tcggcgtgca gctggacggc
     3601 accgagcggc tgcacggagc agcagagagg gaagccgtgt gcctgatcaa gaagacagcg
     3661 tttcagattg ccgaagctgc taatgatgaa aattacttcg tcgactctgc cgggggatct
     3721 gccggaggcg agttcctggc taccacactg gaaagaattg agaagaattt tgtcattacc
     3781 gaccccagac tgcccgacaa tcccatcatc ttcgcctccg actcttttct gcagctgacc
     3841 gagtattcta gagaggagat cctgggaaga aactgcagat tcctgcaggg acctgagaca
     3901 gacagggcta cagtgcggaa aattagagat gctattgaca atcagactga agtcactgtc
     3961 cagctgatca attacactaa gtctggcaag aagttctgga acgtgtttca cctgcaaccc
     4021 atgagagatt ataagggcga tgtgcagtat ttcatcgggg tccagctgga tggcactgag
     4081 aggctgcatg gcgccgctga aagggaggcc gtgtgcctga tcaagaagac agcttttcag
     4141 attgctgaag ctgccaacga cgagaattat tttggatcca ccggatctag ataactgatc
     4201 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     4261 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     4321 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     4381 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag
     4441 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     4501 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     4561 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     4621 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     4681 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     4741 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     4801 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     4861 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     4921 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4981 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     5041 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5101 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     5161 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5221 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5281 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     5341 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     5401 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     5461 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     5521 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     5581 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     5641 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     5701 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5761 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5821 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5881 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     5941 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     6001 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     6061 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     6121 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     6181 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     6241 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     6301 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     6361 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     6421 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     6481 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     6541 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     6601 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     6661 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6721 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6781 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6841 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6901 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6961 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7021 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7081 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7141 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7201 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7261 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7321 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7381 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7441 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     7501 attaccgcca tgcat
//
