LOCUS       sequence-474358-        5357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the I3-01 nanocage subunit N-terminally tagged with 
            mStayGold 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 5357)
  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 5357)
  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_I3
FEATURES             Location/Qualifiers
     source          1..5357
                     /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"
     polyA_signal    2848..2969
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2885..2904)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2939..2958
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2976..3431)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3113..3134)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3325..3344
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3458..3562
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3564..3921
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3564..3755
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3587..3607)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3772..3907
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3956..4750
                     /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             3956..4750
                     /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(4010..4029)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4620..4639
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4938..4957)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4982..5029
                     /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 ggtgggtcag gcggatccgg tggtagcggt ggtagcggtg
     2101 gtagcggtgg tagcggtggt agcgaggaac tctttaagaa gcataaaatc gtagctgtgc
     2161 tcagagcaaa ttcagtagag gaggcgaaga aaaaggccct ggccgttttc ctgggcggag
     2221 tccacctcat cgaaattact tttactgtcc ccgacgcaga tacagtcata aaagagttga
     2281 gttttctcaa agagatgggg gcgatcatcg gtgccgggac ggttacgtca gtcgagcagt
     2341 gccggaaagc agtggagtca ggagcggaat ttatcgtatc tccacatctg gatgaggaaa
     2401 tatctcaatt ctgtaaagaa aaaggtgtct tttacatgcc cggcgttatg acgccaacgg
     2461 agctggtgaa ggccatgaag ctgggacaca cgatactggc tctctttcca ggcgaagtgg
     2521 tcggtcctca attcgtcaag gcaatgaaag ggcctttccc caacgttaaa ttcgtcccga
     2581 ctgggggagt aaacttggac aatgtttgcg aatggtttaa ggcaggggta ctcgctgttg
     2641 gagttggttc cgcattggta aaaggtacgc ctgtcgaagt ggcagaaaag gcaaaggctt
     2701 tcgttgagaa gatccgcgga tgtactgaat aggcgactct agatcataat cagccatacc
     2761 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     2821 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     2881 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2941 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt
     3001 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     3061 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     3121 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     3181 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     3241 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     3301 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     3361 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     3421 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3481 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     3541 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3601 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3661 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3721 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3781 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3841 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3901 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     3961 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4021 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4081 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     4141 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4201 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4261 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4321 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4381 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4441 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     4501 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4561 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4621 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4681 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4741 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4801 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4861 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4921 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     4981 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     5041 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     5101 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     5161 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     5221 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5281 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5341 aagatcaaag gatcttc
//
