LOCUS       sequence-196609-        6858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of iPOLYMER component in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6858)
  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 6858)
  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: YFP-SspBx6
FEATURES             Location/Qualifiers
     source          1..6858
                     /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             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3646..3767
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3683..3702)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3737..3756
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3774..4229)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3911..3932)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4123..4142
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4256..4360
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4362..4719
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4362..4553
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4385..4405)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4570..4705
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4754..5548
                     /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             4754..5548
                     /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(4808..4827)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5418..5437
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5736..5755)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5780..5827
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6156..6744
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6645..6664
                     /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 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccttcggc tacggcctgc agtgcttcgc ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctagcgct ggaggcagcg ctggcggaag ctccccaaag
     1381 aggcctaagc tgctgagaga gtactatgac tggctggtgg ataacagctt caccccctac
     1441 ctggtggtgg acgccacata tctgggcgtg aacgtgcctg tggagtacgt gaaggatggc
     1501 cagatcgtgc tgaatctgag cgcctccgcc accggaaacc tgcagctgac aaatgacttc
     1561 atccagttta atgcccagtt caagggcgtg tccagggagc tgtacatccc aatgggagcc
     1621 gccctggcaa tctatgcaag ggagaacggc gacggcgtga tgtttgagcc tgaagaaatc
     1681 tacgatgagc tgaatatcgg caagctttcc gcaggcgggt ccgcaggggg ctctagccca
     1741 aagcggccca agctgctgcg cgaatattac gattggctgg tggataactc ttttacccca
     1801 tacctggtcg tcgatgctac ctatctggga gtgaacgtgc ccgtggagta tgtcaaagac
     1861 ggacaaatcg tgctgaatct gtctgccagc gccaccggca acctgcagct gaccaacgat
     1921 ttcatccagt ttaatgctca gttcaagggc gtgagcagag aactgtacat cccaatgggc
     1981 gccgccctgg ccatctatgc cagagaaaat ggcgacggcg tgatgttcga gcccgaggag
     2041 atatacgacg agttgaacat tggagtcgac tctgccgggg gatctgccgg aggctcctct
     2101 cctaagaggc caaaactgct gagggaatat tacgactggc tggtcgataa ctccttcacc
     2161 ccttacctgg tcgtcgatgc aacctatctg ggggtgaacg tgccagtgga gtatgtcaaa
     2221 gatggacaga tcgtgctgaa tctgtccgcc tctgccactg gaaacctgca gctgaccaat
     2281 gacttcatcc agtttaacgc acagttcaag ggcgtgagcc gggagctgta catccctatg
     2341 ggcgccgccc tggctatcta tgccagagaa aacggcgacg gcgtgatgtt tgaacccgaa
     2401 gaaatatatg atgaactcaa tataggggga tctagcgctg gaggcagcgc tggcggaagc
     2461 tccccaaaga ggcctaagct gctgagagag tactatgact ggctggtgga taacagcttc
     2521 accccctacc tggtggtgga cgccacatat ctgggcgtga acgtgcctgt ggagtacgtg
     2581 aaggatggcc agatcgtgct gaatctgagc gcctccgcca ccggaaacct gcagctgaca
     2641 aatgacttca tccagtttaa tgcccagttc aagggcgtgt ccagggagct gtacatccca
     2701 atgggagccg ccctggcaat ctatgcaagg gagaacggcg acggcgtgat gtttgagcct
     2761 gaagaaatct acgatgagct gaatatcggc aagctttccg caggcgggtc cgcagggggc
     2821 tctagcccaa agcggcccaa gctgctgcgc gaatattacg attggctggt ggataactct
     2881 tttaccccat acctggtcgt cgatgctacc tatctgggag tgaacgtgcc cgtggagtat
     2941 gtcaaagacg gacaaatcgt gctgaatctg tctgccagcg ccaccggcaa cctgcagctg
     3001 accaacgatt tcatccagtt taatgctcag ttcaagggcg tgagcagaga actgtacatc
     3061 ccaatgggcg ccgccctggc catctatgcc agagaaaatg gcgacggcgt gatgttcgag
     3121 cccgaggaga tatacgacga gttgaacatt ggagtcgact ctgccggggg atctgccgga
     3181 ggctcctctc ctaagaggcc aaaactgctg agggaatatt acgactggct ggtcgataac
     3241 tccttcaccc cttacctggt cgtcgatgca acctatctgg gggtgaacgt gccagtggag
     3301 tatgtcaaag atggacagat cgtgctgaat ctgtccgcct ctgccactgg aaacctgcag
     3361 ctgaccaatg acttcatcca gtttaacgca cagttcaagg gcgtgagccg ggagctgtac
     3421 atccctatgg gcgccgccct ggctatctat gccagagaaa acggcgacgg cgtgatgttt
     3481 gaacccgaag aaatatatga tgaactcaat atagggggat ccaccggatc tagataactg
     3541 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3601 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3661 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3721 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     3781 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     3841 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     3901 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3961 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     4021 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     4081 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     4141 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     4201 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     4261 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4321 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     4381 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4441 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4501 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4561 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4621 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     4681 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     4741 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4801 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4861 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4921 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4981 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     5041 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     5101 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     5161 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     5221 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5281 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5341 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5401 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5461 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     5521 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     5581 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5641 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5701 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     5761 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     5821 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     5881 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     5941 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     6001 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6061 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6121 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6181 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6241 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6301 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6361 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6421 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6481 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6541 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6601 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6661 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6721 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6781 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6841 cgtattaccg ccatgcat
//
