LOCUS       sequence-196618-        7517 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 7517)
  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 7517)
  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-YFP-SspBx6
FEATURES             Location/Qualifiers
     source          1..7517
                     /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             join(1272..1274,1275..1277,1278..1988)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1317..1338)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1578..1597)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1925..1946
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4305..4426
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4342..4361)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4396..4415
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4433..4888)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4570..4591)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4782..4801
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4915..5019
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5021..5378
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5021..5212
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5044..5064)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5229..5364
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5413..6207
                     /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             5413..6207
                     /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(5467..5486)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6077..6096
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6395..6414)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6439..6486
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6815..7403
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7304..7323
                     /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 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1321 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1381 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     1441 cctggcccac cctcgtgacc accttcggct acggcctgca gtgcttcgcc cgctaccccg
     1501 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1561 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1621 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1681 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     1741 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     1801 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1861 ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac gagaagcgcg
     1921 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1981 tgtacaagtc cggactcaga tctagcgctg gaggcagcgc tggcggaagc tccccaaaga
     2041 ggcctaagct gctgagagag tactatgact ggctggtgga taacagcttc accccctacc
     2101 tggtggtgga cgccacatat ctgggcgtga acgtgcctgt ggagtacgtg aaggatggcc
     2161 agatcgtgct gaatctgagc gcctccgcca ccggaaacct gcagctgaca aatgacttca
     2221 tccagtttaa tgcccagttc aagggcgtgt ccagggagct gtacatccca atgggagccg
     2281 ccctggcaat ctatgcaagg gagaacggcg acggcgtgat gtttgagcct gaagaaatct
     2341 acgatgagct gaatatcggc aagctttccg caggcgggtc cgcagggggc tctagcccaa
     2401 agcggcccaa gctgctgcgc gaatattacg attggctggt ggataactct tttaccccat
     2461 acctggtcgt cgatgctacc tatctgggag tgaacgtgcc cgtggagtat gtcaaagacg
     2521 gacaaatcgt gctgaatctg tctgccagcg ccaccggcaa cctgcagctg accaacgatt
     2581 tcatccagtt taatgctcag ttcaagggcg tgagcagaga actgtacatc ccaatgggcg
     2641 ccgccctggc catctatgcc agagaaaatg gcgacggcgt gatgttcgag cccgaggaga
     2701 tatacgacga gttgaacatt ggagtcgact ctgccggggg atctgccgga ggctcctctc
     2761 ctaagaggcc aaaactgctg agggaatatt acgactggct ggtcgataac tccttcaccc
     2821 cttacctggt cgtcgatgca acctatctgg gggtgaacgt gccagtggag tatgtcaaag
     2881 atggacagat cgtgctgaat ctgtccgcct ctgccactgg aaacctgcag ctgaccaatg
     2941 acttcatcca gtttaacgca cagttcaagg gcgtgagccg ggagctgtac atccctatgg
     3001 gcgccgccct ggctatctat gccagagaaa acggcgacgg cgtgatgttt gaacccgaag
     3061 aaatatatga tgaactcaat atagggggat ctagcgctgg aggcagcgct ggcggaagct
     3121 ccccaaagag gcctaagctg ctgagagagt actatgactg gctggtggat aacagcttca
     3181 ccccctacct ggtggtggac gccacatatc tgggcgtgaa cgtgcctgtg gagtacgtga
     3241 aggatggcca gatcgtgctg aatctgagcg cctccgccac cggaaacctg cagctgacaa
     3301 atgacttcat ccagtttaat gcccagttca agggcgtgtc cagggagctg tacatcccaa
     3361 tgggagccgc cctggcaatc tatgcaaggg agaacggcga cggcgtgatg tttgagcctg
     3421 aagaaatcta cgatgagctg aatatcggca agctttccgc aggcgggtcc gcagggggct
     3481 ctagcccaaa gcggcccaag ctgctgcgcg aatattacga ttggctggtg gataactctt
     3541 ttaccccata cctggtcgtc gatgctacct atctgggagt gaacgtgccc gtggagtatg
     3601 tcaaagacgg acaaatcgtg ctgaatctgt ctgccagcgc caccggcaac ctgcagctga
     3661 ccaacgattt catccagttt aatgctcagt tcaagggcgt gagcagagaa ctgtacatcc
     3721 caatgggcgc cgccctggcc atctatgcca gagaaaatgg cgacggcgtg atgttcgagc
     3781 ccgaggagat atacgacgag ttgaacattg gagtcgactc tgccggggga tctgccggag
     3841 gctcctctcc taagaggcca aaactgctga gggaatatta cgactggctg gtcgataact
     3901 ccttcacccc ttacctggtc gtcgatgcaa cctatctggg ggtgaacgtg ccagtggagt
     3961 atgtcaaaga tggacagatc gtgctgaatc tgtccgcctc tgccactgga aacctgcagc
     4021 tgaccaatga cttcatccag tttaacgcac agttcaaggg cgtgagccgg gagctgtaca
     4081 tccctatggg cgccgccctg gctatctatg ccagagaaaa cggcgacggc gtgatgtttg
     4141 aacccgaaga aatatatgat gaactcaata tagggggatc caccggatct agataactga
     4201 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     4261 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     4321 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4381 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta
     4441 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     4501 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     4561 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     4621 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     4681 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     4741 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4801 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4861 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4921 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4981 ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc
     5041 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     5101 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     5161 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     5221 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5281 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     5341 agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga
     5401 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     5461 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     5521 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5581 ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5641 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5701 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5761 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5821 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5881 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5941 agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     6001 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     6061 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     6121 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     6181 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     6241 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     6301 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     6361 tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg
     6421 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt
     6481 ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac
     6541 cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg
     6601 gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt
     6661 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6721 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6781 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6841 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6901 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6961 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     7021 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     7081 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     7141 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     7201 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     7261 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     7321 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     7381 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     7441 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     7501 gtattaccgc catgcat
//
