LOCUS       sequence-474988-        6014 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the I3-01 nanocage subunit N-terminally tagged with two 
            copies of mStayGold in mammalian cells. Assembles into nanocages 
            tagged with 120 FPs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6014)
  AUTHORS   Viola G, Ibrahim YW, Jacobs KA, Lemiere J, Kutys ML, Wittmann T
  TITLE     Quantitative comparison of fluorescent proteins using protein 
            nanocages in live cells.
  JOURNAL   J Cell Sci. 2025 May 15;138(10):jcs263858. doi: 10.1242/jcs.263858. 
            Epub 2025 May 21.
  PUBMED    40326462
REFERENCE   2  (bases 1 to 6014)
  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: "J Cell 
            Sci."; date: "2025-05-15"; volume: "138(10):jcs263858. doi"; pages: 
            " 10.1242/jcs.263858. Epub 2025 May 21"
COMMENT     Sequence Label: pmStayGold_mStayGold_I3
FEATURES             Location/Qualifiers
     source          1..6014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        764..1067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1068..1271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1222..1242
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    1312..1368
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      1377..1386
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1383..2060
                     /codon_start=1
                     /product="bright and highly photostable monomeric green 
                     fluorescent protein (Ando et al., 2023)"
                     /label=mStayGold(J)
                     /note="includes multiple mutations plus an N-terminal 
                     iinker sequence"
                     /translation="MVSTGEELFTGVVPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKG
                     KYVCTSGKLPMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQYKGDGS
                     IHAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMDVSLPNEVQHIPIDDGVECTVTL
                     QYPLLSDESKCVEAYQNTIIKPLHNQPAPDVPFHWIRKQYTQSKDDTEERDHIIQSETL
                     EAHL"
     CDS             2070..2744
                     /codon_start=1
                     /product="bright and highly photostable monomeric green 
                     fluorescent protein (Ando et al., 2023)"
                     /label=mStayGold(J)
                     /note="includes multiple mutations plus an N-terminal 
                     iinker sequence"
                     /translation="VSTGEELFTGVVPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKGK
                     YVCTSGKLPMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQYKGDGSI
                     HAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMDVSLPNEVQHIPIDDGVECTVTLQ
                     YPLLSDESKCVEAYQNTIIKPLHNQPAPDVPFHWIRKQYTQSKDDTEERDHIIQSETLE
                     AHL"
     polyA_signal    3505..3626
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3542..3561)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3596..3615
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3633..4088)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3770..3791)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3982..4001
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4115..4219
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4221..4578
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4221..4412
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4244..4264)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4429..4564
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4613..5407
                     /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             4613..5407
                     /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(4667..4686)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5277..5296
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5595..5614)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5639..5686
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
      721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1261 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact cagatctcga
     1321 gctcaagctt cgaattctgc agtcgacggt accgcgggcc cgggatccaa ccggtcgcca
     1381 ccatggtgtc tacaggcgag gagctgttta ccggcgtggt gcccttcaag ttccagctga
     1441 agggcaccat caacggcaag agcttcaccg tggaaggcga gggcgagggc aatagccacg
     1501 agggcagcca caaaggcaag tacgtgtgca ccagcggcaa actgccaatg tcttgggccg
     1561 ccctgggaac tagcttcggc tatggcatga agtactacac caagtacccc agcggcctga
     1621 agaactggtt ccacgaggtg atgcccgagg gcttcaccta cgacagacac atccagtaca
     1681 agggcgacgg cagcatccac gccaagcacc agcacttcat gaagaacggc acctaccaca
     1741 acatcgtgga gttcaccggc caggacttca aggagaacag ccccgtgctg accggcgaca
     1801 tggacgtgag cctgcccaac gaggtgcagc acatccccat agatgacggc gtggagtgca
     1861 cagtgaccct gcagtaccct ctgctgagcg acgagagcaa gtgcgtggag gcctaccaga
     1921 acaccatcat caagcccctg cacaatcagc cagcccccga tgtgccattc cactggatca
     1981 gaaagcagta cacccagagc aaggacgaca ccgaggagag agaccacatc atccagagcg
     2041 agaccctgga ggcccacctg ggaggcagcg tgtctacagg cgaggagctg tttaccggcg
     2101 tggtgccctt caagttccag ctgaagggca ccatcaacgg caagagcttc accgtggaag
     2161 gcgagggcga gggcaatagc cacgagggca gccacaaagg caagtacgtg tgcaccagcg
     2221 gcaaactgcc aatgtcttgg gccgccctgg gaactagctt cggctatggc atgaagtact
     2281 acaccaagta ccccagcggc ctgaagaact ggttccacga ggtgatgccc gagggcttca
     2341 cctacgacag acacatccag tacaagggcg acggcagcat ccacgccaag caccagcact
     2401 tcatgaagaa cggcacctac cacaacatcg tggagttcac cggccaggac ttcaaggaga
     2461 acagccccgt gctgaccggc gacatggacg tgagcctgcc caacgaggtg cagcacatcc
     2521 ccatagatga cggcgtggag tgcacagtga ccctgcagta ccctctgctg agcgacgaga
     2581 gcaagtgcgt ggaggcctac cagaacacca tcatcaagcc cctgcacaat cagccagccc
     2641 ccgatgtgcc attccactgg atcagaaagc agtacaccca gagcaaggac gacaccgagg
     2701 agagagacca catcatccag agcgagaccc tggaggccca cctgggtggg tcaggcggat
     2761 ccggtggtag cggtggtagc gaggaactct ttaagaagca taaaatcgta gctgtgctca
     2821 gagcaaattc agtagaggag gcgaagaaaa aggccctggc cgttttcctg ggcggagtcc
     2881 acctcatcga aattactttt actgtccccg acgcagatac agtcataaaa gagttgagtt
     2941 ttctcaaaga gatgggggcg atcatcggtg ccgggacggt tacgtcagtc gagcagtgcc
     3001 ggaaagcagt ggagtcagga gcggaattta tcgtatctcc acatctggat gaggaaatat
     3061 ctcaattctg taaagaaaaa ggtgtctttt acatgcccgg cgttatgacg ccaacggagc
     3121 tggtgaaggc catgaagctg ggacacacga tactggctct ctttccaggc gaagtggtcg
     3181 gtcctcaatt cgtcaaggca atgaaagggc ctttccccaa cgttaaattc gtcccgactg
     3241 ggggagtaaa cttggacaat gtttgcgaat ggtttaaggc aggggtactc gctgttggag
     3301 ttggttccgc attggtaaaa ggtacgcctg tcgaagtggc agaaaaggca aaggctttcg
     3361 ttgagaagat ccgcggatgt actgaatagg cgactctaga tcataatcag ccataccaca
     3421 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     3481 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     3541 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3601 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
     3661 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     3721 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     3781 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     3841 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     3901 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     3961 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     4021 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     4081 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4141 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4201 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     4261 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4321 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4381 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4441 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4501 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4561 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     4621 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4681 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4741 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     4801 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4861 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4921 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4981 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     5041 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     5101 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     5161 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     5221 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     5281 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     5341 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     5401 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     5461 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5521 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     5581 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     5641 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     5701 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     5761 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     5821 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     5881 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5941 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6001 atcaaaggat cttc
//
