LOCUS       sequence-474350-        5369 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the I3-01 nanocage subunit N-terminally tagged with one 
            mEmerald in mammalian cells. Assembles into nanocages tagged with 60
            mEmerald FPs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5369)
  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 5369)
  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_I3
FEATURES             Location/Qualifiers
     source          1..5369
                     /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"
     primer_bind     complement(1428..1449)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1689..1708)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2036..2057
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2860..2981
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2897..2916)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2951..2970
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2988..3443)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3125..3146)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3337..3356
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3470..3574
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3576..3933
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3576..3767
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3599..3619)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3784..3919
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3968..4762
                     /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             3968..4762
                     /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(4022..4041)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4632..4651
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4950..4969)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4994..5041
                     /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 gtggtagcgg tggtagcggt ggtagcggtg gtagcgagga actctttaag aagcataaaa
     2161 tcgtagctgt gctcagagca aattcagtag aggaggcgaa gaaaaaggcc ctggccgttt
     2221 tcctgggcgg agtccacctc atcgaaatta cttttactgt ccccgacgca gatacagtca
     2281 taaaagagtt gagttttctc aaagagatgg gggcgatcat cggtgccggg acggttacgt
     2341 cagtcgagca gtgccggaaa gcagtggagt caggagcgga atttatcgta tctccacatc
     2401 tggatgagga aatatctcaa ttctgtaaag aaaaaggtgt cttttacatg cccggcgtta
     2461 tgacgccaac ggagctggtg aaggccatga agctgggaca cacgatactg gctctctttc
     2521 caggcgaagt ggtcggtcct caattcgtca aggcaatgaa agggcctttc cccaacgtta
     2581 aattcgtccc gactggggga gtaaacttgg acaatgtttg cgaatggttt aaggcagggg
     2641 tactcgctgt tggagttggt tccgcattgg taaaaggtac gcctgtcgaa gtggcagaaa
     2701 aggcaaaggc tttcgttgag aagatccgcg gatgtactga ataggcgact ctagatcata
     2761 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     2821 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     2881 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     2941 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
     3001 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     3061 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     3121 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     3181 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     3241 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     3301 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     3361 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     3421 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3481 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3541 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     3601 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3661 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3721 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3781 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3841 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3901 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     3961 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4021 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4081 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4141 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4201 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4261 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4321 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4381 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4441 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     4501 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4561 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4621 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4681 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4741 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4801 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4861 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4921 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     4981 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     5041 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     5101 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     5161 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     5221 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5281 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5341 gaccccgtag aaaagatcaa aggatcttc
//
