LOCUS       sequence-189396-        5571 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 5571)
  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 5571)
  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..5571
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             613..891
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISK"
     CDS             625..894
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS
                     FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQ"
     regulatory      910..919
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             916..1623
                     /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(1061..1079)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1329..1348
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2359..2480
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2396..2415)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2450..2469
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2487..2942)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2624..2645)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2836..2855
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2969..3073
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3075..3432
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3075..3266
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3098..3118)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3283..3418
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3467..4261
                     /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             3467..4261
                     /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(3521..3540)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4131..4150
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4449..4468)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4493..4540
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4869..5457
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5358..5377
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcggat
      601 ccggccacca tgatcctctg gcatgagatg tggcatgaag gcctggaaga ggcatctcgt
      661 ttgtactttg gggaaaggaa cgtgaaaggc atgtttgagg tgctggagcc cttgcatgct
      721 atgatggaac ggggccccca gactctgaag gaaacatcct ttaatcaggc ctatggtcga
      781 gatttaatgg aggcccaaga gtggtgcagg aagtacatga aatcagggaa tgtcaaggac
      841 ctcacccaag cctgggacct ctattatcat gtgttccgac gaatctcaaa gcaggccggc
      901 ccaccggtcg ccaccatggt gagcaagggc gaggaggata acatggccat catcaaggag
      961 ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag
     1021 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     1081 ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag
     1141 gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc
     1201 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac
     1261 tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc
     1321 tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg
     1381 taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc
     1441 ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc
     1501 ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc
     1561 gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac
     1621 aagtccggac tcagatctcg agccatgacg accgcgtcca cctcgcaggt gcgccagaac
     1681 taccaccagg actcagaggc cgccatcaac cgccagatca acctggagct ctacgcctcc
     1741 tacgtttacc tgtccatgtc ttactacttt gaccgcgatg atgtggcttt gaagaacttt
     1801 gccaaatact ttcttcacca atctcatgag gagagggaac atgctgagaa actgatgaag
     1861 ctgcagaacc aacgaggtgg ccgaatcttc cttcaggata tcaagaaacc agactgtgat
     1921 gactgggaga gcgggctgaa tgcaatggag tgtgcattac atttggaaaa aaatgtgaat
     1981 cagtcactac tggaactgca caaactggcc actgacaaaa atgaccccca tttgtgtgac
     2041 ttcattgaga cacattacct gaatgagcag gtgaaagcca tcaaagaatt gggtgaccac
     2101 gtgaccaact tgcgcaagat gggagcgccc gaatctggct tggcggaata tctctttgac
     2161 aagcacaccc tgggagacag tgataatgaa agctaaccga attctgcagt cgacggtacc
     2221 gcgggcccgg gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta
     2281 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     2341 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     2401 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     2461 aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     2521 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     2581 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     2641 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     2701 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     2761 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     2821 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     2881 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     2941 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3001 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3061 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     3121 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3181 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3241 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3301 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3361 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3421 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3481 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3541 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3601 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     3661 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3721 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3781 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3841 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3901 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3961 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     4021 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4081 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4141 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4201 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4261 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4321 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4381 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     4441 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     4501 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     4561 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     4621 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     4681 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     4741 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4801 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4861 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4921 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4981 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     5041 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5101 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5161 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5221 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5281 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5341 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5401 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5461 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     5521 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
