LOCUS       sequence-194021-        5565 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Part of FerriTag expression system. Encodes human Ferritin with an 
            FRB domain and mCherry tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5565)
  AUTHORS   Clarke NI, Royle SJ
  TITLE     FerriTag is a new genetically-encoded inducible tag for correlative 
            light-electron microscopy.
  JOURNAL   Nat Commun. 2018 Jul 4;9(1):2604. doi: 10.1038/s41467-018-04993-0.
  PUBMED    29973588
REFERENCE   2  (bases 1 to 5565)
  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: "Nat 
            Commun."; date: "2018-07-4"; volume: "9(1):2604. doi"; pages: " 
            10.1038/s41467-018-04993-0"
COMMENT     Sequence Label: FRB-mCherry-FTH1
FEATURES             Location/Qualifiers
     source          1..5565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        62..365
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        366..569
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     520..540
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             608..886
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISK"
     CDS             620..889
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS
                     FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQ"
     regulatory      905..914
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             911..1618
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1056..1074)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1324..1343
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2354..2475
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2391..2410)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2445..2464
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2482..2937)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2619..2640)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2831..2850
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2964..3068
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3070..3427
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3070..3261
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3093..3113)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3278..3413
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3462..4256
                     /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             3462..4256
                     /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(3516..3535)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4126..4145
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4444..4463)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4488..4535
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4864..5452
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5353..5372
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
       61 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      121 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      181 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      241 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      301 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      361 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      421 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      481 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      541 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcggc
      601 caccatgatc ctctggcatg agatgtggca tgaaggcctg gaagaggcat ctcgtttgta
      661 ctttggggaa aggaacgtga aaggcatgtt tgaggtgctg gagcccttgc atgctatgat
      721 ggaacggggc ccccagactc tgaaggaaac atcctttaat caggcctatg gtcgagattt
      781 aatggaggcc caagagtggt gcaggaagta catgaaatca gggaatgtca aggacctcac
      841 ccaagcctgg gacctctatt atcatgtgtt ccgacgaatc tcaaagcagg ccggcccacc
      901 ggtcgccacc atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat
      961 gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga
     1021 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg
     1081 ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta
     1141 cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa
     1201 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc
     1261 cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga
     1321 cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc
     1381 cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca
     1441 ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc
     1501 ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga
     1561 acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagtc
     1621 cggactcaga tctcgagcca tgacgaccgc gtccacctcg caggtgcgcc agaactacca
     1681 ccaggactca gaggccgcca tcaaccgcca gatcaacctg gagctctacg cctcctacgt
     1741 ttacctgtcc atgtcttact actttgaccg cgatgatgtg gctttgaaga actttgccaa
     1801 atactttctt caccaatctc atgaggagag ggaacatgct gagaaactga tgaagctgca
     1861 gaaccaacga ggtggccgaa tcttccttca ggatatcaag aaaccagact gtgatgactg
     1921 ggagagcggg ctgaatgcaa tggagtgtgc attacatttg gaaaaaaatg tgaatcagtc
     1981 actactggaa ctgcacaaac tggccactga caaaaatgac ccccatttgt gtgacttcat
     2041 tgagacacat tacctgaatg agcaggtgaa agccatcaaa gaattgggtg accacgtgac
     2101 caacttgcgc aagatgggag cgcccgaatc tggcttggcg gaatatctct ttgacaagca
     2161 caccctggga gacagtgata atgaaagcta accgaattct gcagtcgacg gtaccgcggg
     2221 cccgggatcc accggatcta gataactgat cataatcagc cataccacat ttgtagaggt
     2281 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     2341 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     2401 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     2461 catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2521 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     2581 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     2641 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     2701 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     2761 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     2821 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     2881 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     2941 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3001 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3061 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     3121 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3181 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3241 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3301 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3361 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3421 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3481 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3541 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3601 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     3661 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3721 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3781 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3841 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3901 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     3961 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     4021 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4081 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4141 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4201 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4261 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4321 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4381 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     4441 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     4501 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     4561 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     4621 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     4681 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     4741 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4801 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4861 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4921 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4981 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5041 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5101 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5161 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5221 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5281 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5341 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5401 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5461 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5521 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgca
//
