LOCUS       sequence-474987-        5354 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the I3-01 nanocage subunit N-terminally tagged with 
            mCherry in mammalian cells. Assembles into nanocages tagged with 60 
            FPs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5354)
  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 5354)
  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: pmCherry_I3
FEATURES             Location/Qualifiers
     source          1..5354
                     /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"
     polyA_signal    2845..2966
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2882..2901)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2936..2955
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2973..3428)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3110..3131)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3322..3341
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3455..3559
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3561..3918
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3561..3752
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3584..3604)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3769..3904
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3953..4747
                     /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             3953..4747
                     /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(4007..4026)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4617..4636
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4935..4954)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4979..5026
                     /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 aaaaggcgag gaggacaata tggcgattat caaggaattt atgcgcttta
     1441 aagtccatat ggaggggtcc gttaacgggc acgagttcga gatcgaaggt gaaggtgagg
     1501 gcaggcccta cgaaggcact cagaccgcta agctgaaggt gactaaagga ggccccctcc
     1561 cctttgcctg ggacatactc agccctcagt ttatgtacgg gtcaaaggcc tacgtcaagc
     1621 accctgcaga tataccagac tacttgaaat tatcatttcc cgaggggttt aaatgggaga
     1681 gagttatgaa ttttgaggat ggcggcgtag ttactgtaac acaggatagt agcctgcagg
     1741 atggggaatt tatttacaaa gtgaaattac gagggactaa ctttccttcc gatggaccag
     1801 tgatgcagaa aaagactatg ggttgggagg cgtcctcaga aagaatgtat cctgaggatg
     1861 gtgcactcaa aggagagatc aagcaaaggc tgaagctgaa ggatggtgga cattatgacg
     1921 ccgaagttaa aaccacatac aaggcaaaga aaccggtaca gttgccgggt gcatacaatg
     1981 tgaatattaa actggacatc acctcacaca atgaagacta caccattgtc gaacagtatg
     2041 agcgtgctga aggacgacat agtactggcg gatccggtgg tagcggtggt agcggtggta
     2101 gcggtggtag cggtggtagc gaggaactct ttaagaagca taaaatcgta gctgtgctca
     2161 gagcaaattc agtagaggag gcgaagaaaa aggccctggc cgttttcctg ggcggagtcc
     2221 acctcatcga aattactttt actgtccccg acgcagatac agtcataaaa gagttgagtt
     2281 ttctcaaaga gatgggggcg atcatcggtg ccgggacggt tacgtcagtc gagcagtgcc
     2341 ggaaagcagt ggagtcagga gcggaattta tcgtatctcc acatctggat gaggaaatat
     2401 ctcaattctg taaagaaaaa ggtgtctttt acatgcccgg cgttatgacg ccaacggagc
     2461 tggtgaaggc catgaagctg ggacacacga tactggctct ctttccaggc gaagtggtcg
     2521 gtcctcaatt cgtcaaggca atgaaagggc ctttccccaa cgttaaattc gtcccgactg
     2581 ggggagtaaa cttggacaat gtttgcgaat ggtttaaggc aggggtactc gctgttggag
     2641 ttggttccgc attggtaaaa ggtacgcctg tcgaagtggc agaaaaggca aaggctttcg
     2701 ttgagaagat ccgcggatgt actgaatagg cgactctaga tcataatcag ccataccaca
     2761 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     2821 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     2881 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     2941 ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa
     3001 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     3061 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     3121 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     3181 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     3241 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     3301 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     3361 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     3421 gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3481 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3541 atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     3601 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3661 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3721 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3781 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3841 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3901 ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga
     3961 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4021 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4081 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga
     4141 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4201 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4261 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4321 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4381 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4441 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga
     4501 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4561 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4621 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4681 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4741 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4801 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4861 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct
     4921 agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg
     4981 gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt
     5041 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     5101 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     5161 gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt
     5221 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5281 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5341 atcaaaggat cttc
//
