LOCUS       sequence-474359-        5330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the I3-01 nanocage subunit N-terminally tagged with 
            mScarlet3 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 5330)
  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 5330)
  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: pmScarlet3_I3
FEATURES             Location/Qualifiers
     source          1..5330
                     /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..2069
                     /codon_start=1
                     /product="bright cysteine-free monomeric red fluorescent 
                     protein with fast and complete maturation (Gadella et al., 
                     2023)"
                     /label=mScarlet3
                     /translation="MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LRVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYWKQSFPEGFKWERVMNFEDGGA
                     VSVAQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDVVLKGDIKM
                     ALRLKDGGRYLADFKTTYRAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     polyA_signal    2821..2942
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2858..2877)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2912..2931
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2949..3404)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3086..3107)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3298..3317
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3431..3535
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3537..3894
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3537..3728
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3560..3580)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3745..3880
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3929..4723
                     /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             3929..4723
                     /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(3983..4002)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4593..4612
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4911..4930)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4955..5002
                     /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 ccatggattc aaccgaagcc gtgatcaaag aatttatgcg ttttaaggtc cacatggaag
     1441 gcagcatgaa tggccacgaa tttgaaatcg aaggtgaggg tgagggtcgg ccttacgagg
     1501 gtactcagac cgctaagtta agagtgacca agggtggccc attgccattc tcctgggaca
     1561 tcttatctcc ccaattcatg tacgggtccc gggcgtttac caaacaccct gccgatattc
     1621 ctgactattg gaagcagtct ttccccgagg gatttaaatg ggagcgcgtg atgaactttg
     1681 aagatggcgg agccgtgtcc gtggctcagg acacttcact ggaggatggc accttaatct
     1741 acaaggttaa gctcagaggg actaacttcc ccccggacgg gcccgtaatg cagaaaaaga
     1801 ccatggggtg ggaagcctcc accgagcgat tgtaccctga agatgtggtg ctgaagggag
     1861 atattaagat ggctttaaga ctcaaggatg gcggccgcta tcttgccgac ttcaagacca
     1921 cttaccgtgc taaaaaacca gtgcagatgc ccggcgcctt caacatcgac cggaaactgg
     1981 atatcaccag tcacaacgag gattatacag tggtagagca gtacgagagg tccgtggcaa
     2041 gacattctac cggcggatcc ggtggtagcg gtggtagcgg tggtagcggt ggtagcgagg
     2101 aactctttaa gaagcataaa atcgtagctg tgctcagagc aaattcagta gaggaggcga
     2161 agaaaaaggc cctggccgtt ttcctgggcg gagtccacct catcgaaatt acttttactg
     2221 tccccgacgc agatacagtc ataaaagagt tgagttttct caaagagatg ggggcgatca
     2281 tcggtgccgg gacggttacg tcagtcgagc agtgccggaa agcagtggag tcaggagcgg
     2341 aatttatcgt atctccacat ctggatgagg aaatatctca attctgtaaa gaaaaaggtg
     2401 tcttttacat gcccggcgtt atgacgccaa cggagctggt gaaggccatg aagctgggac
     2461 acacgatact ggctctcttt ccaggcgaag tggtcggtcc tcaattcgtc aaggcaatga
     2521 aagggccttt ccccaacgtt aaattcgtcc cgactggggg agtaaacttg gacaatgttt
     2581 gcgaatggtt taaggcaggg gtactcgctg ttggagttgg ttccgcattg gtaaaaggta
     2641 cgcctgtcga agtggcagaa aaggcaaagg ctttcgttga gaagatccgc ggatgtactg
     2701 aataggcgac tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta
     2761 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     2821 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2881 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2941 taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     3001 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     3061 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     3121 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     3181 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     3241 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3301 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3361 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     3421 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3481 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     3541 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3601 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3661 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3721 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3781 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     3841 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     3901 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3961 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4021 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4081 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     4141 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4201 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4261 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4321 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4381 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4441 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4501 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4561 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4621 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4681 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4741 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4801 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4861 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     4921 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     4981 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     5041 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     5101 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     5161 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     5221 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5281 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
