LOCUS       sequence-474362-        6098 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the I3-01 nanocage subunit N-terminally tagged with two 
            tandem mEmeralds in mammalian cells. Assembles into nanocages tagged
            with 120 mEmerald FPs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6098)
  AUTHORS   Dema A, Charafeddine RA, van Haren J, Rahgozar S, Viola G, Jacobs 
            KA, Kutys ML, Wittmann T
  TITLE     Doublecortin reinforces microtubules to promote growth cone advance 
            in soft environments.
  JOURNAL   Curr Biol. 2024 Nov 21:S0960-9822(24)01458-1. doi: 
            10.1016/j.cub.2024.10.063.
  PUBMED    39626666
REFERENCE   2  (bases 1 to 6098)
  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: "Curr Biol. 
            2024 Nov 21:S0960-9822(24)01458-1. doi: 10.1016/j.cub.2024.10.063."
COMMENT     Sequence Label: pmEmerald_mEmerald_I3
FEATURES             Location/Qualifiers
     source          1..6098
                     /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             join(1383..1385,1386..1388,1389..2099)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2115..2117,2118..2828)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIKV
                     NFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     polyA_signal    3589..3710
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3626..3645)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3680..3699
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3717..4172)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3854..3875)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4066..4085
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4199..4303
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4305..4662
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4305..4496
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4328..4348)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4513..4648
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4697..5491
                     /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             4697..5491
                     /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(4751..4770)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5361..5380
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5679..5698)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5723..5770
                     /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 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     1441 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     1501 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     1561 ccctcgtgac caccttgacc tacggcgtgc agtgcttcgc ccgctacccc gaccacatga
     1621 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     1681 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     1741 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1801 acaagctgga gtacaactac aacagccaca aggtctatat caccgccgac aagcagaaga
     1861 acggcatcaa ggtgaacttc aagacccgcc acaacatcga ggacggcagc gtgcagctcg
     1921 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     1981 actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg
     2041 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg
     2101 gtggtggtgg tagcgtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     2161 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     2221 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     2281 cctggcccac cctcgtgacc accttgacct acggcgtgca gtgcttcgcc cgctaccccg
     2341 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     2401 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     2461 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     2521 tcctggggca caagctggag tacaactaca acagccacaa ggtctatatc accgccgaca
     2581 agcagaagaa cggcatcaag gtgaacttca agacccgcca caacatcgag gacggcagcg
     2641 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     2701 ccgacaacca ctacctgagc acccagtcca agctgagcaa agaccccaac gagaagcgcg
     2761 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     2821 tgtacaaggg tggtagcggt ggtagcggtg gtagcggtgg tagcgaggaa ctctttaaga
     2881 agcataaaat cgtagctgtg ctcagagcaa attcagtaga ggaggcgaag aaaaaggccc
     2941 tggccgtttt cctgggcgga gtccacctca tcgaaattac ttttactgtc cccgacgcag
     3001 atacagtcat aaaagagttg agttttctca aagagatggg ggcgatcatc ggtgccggga
     3061 cggttacgtc agtcgagcag tgccggaaag cagtggagtc aggagcggaa tttatcgtat
     3121 ctccacatct ggatgaggaa atatctcaat tctgtaaaga aaaaggtgtc ttttacatgc
     3181 ccggcgttat gacgccaacg gagctggtga aggccatgaa gctgggacac acgatactgg
     3241 ctctctttcc aggcgaagtg gtcggtcctc aattcgtcaa ggcaatgaaa gggcctttcc
     3301 ccaacgttaa attcgtcccg actgggggag taaacttgga caatgtttgc gaatggttta
     3361 aggcaggggt actcgctgtt ggagttggtt ccgcattggt aaaaggtacg cctgtcgaag
     3421 tggcagaaaa ggcaaaggct ttcgttgaga agatccgcgg atgtactgaa taggcgactc
     3481 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     3541 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     3601 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     3661 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat
     3721 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     3781 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     3841 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     3901 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     3961 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     4021 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     4081 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     4141 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     4201 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4261 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     4321 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4381 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4441 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4501 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4561 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4621 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     4681 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4741 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4801 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4861 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4921 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4981 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5041 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5101 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5161 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5221 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5281 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5341 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5401 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5461 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     5521 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5581 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5641 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     5701 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     5761 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     5821 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     5881 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     5941 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6001 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6061 tgagcgtcag accccgtaga aaagatcaaa ggatcttc
//
