LOCUS       sequence-407767-        5633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TOM20-ECFP-FIREmate in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5633)
  AUTHORS   Bottone S, Joliot O, Cakil ZV, El Hajji L, Rakotoarison LM, 
            Boncompain G, Perez F, Gautier A
  TITLE     A fluorogenic chemically induced dimerization technology for 
            controlling, imaging and sensing protein proximity.
  JOURNAL   Nat Methods. 2023 Aug 28. doi: 10.1038/s41592-023-01988-8.
  PUBMED    37640938
REFERENCE   2  (bases 1 to 5633)
  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 
            Methods. 2023 Aug 28. doi: 10.1038/s41592-023-01988-8."
COMMENT     Sequence Label: pAG1159 TOM20-ECFP-FIREmate
FEATURES             Location/Qualifiers
     source          1..5633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(146..503)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      160..295
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        312..503
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     460..480
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(505..609)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      636..1091
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(723..742)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     933..954
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1098..1219
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1109..1128)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1163..1182
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(1805..2515,2516..2518,2519..2521))
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1847..1868)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2196..2215
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2455..2476
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2650..2659
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2703..2906)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2732..2752)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2907..3210
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3385..3973)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3465..3484)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4302..4349
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4374..4393
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4581..5375)
                     /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             complement(4581..5375)
                     /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(4692..4711)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     promoter        complement(5410..5606)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        complement(5418..5628)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5424..5559
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
       61 ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga
      121 tcctcatcct gtctcttgat cgatctttgc aaaagcctag gcctccaaaa aagcctcctc
      181 actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa
      241 ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg
      301 gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc
      361 ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt
      421 gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc
      481 acagctggtt ctttccgcct caggactctt cctttttcaa tattattgaa gcatttatca
      541 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
      601 ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag
      661 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
      721 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
      781 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
      841 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
      901 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
      961 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     1021 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     1081 gcttacaatt tacgccttaa gatacattga tgagtttgga caaaccacaa ctagaatgca
     1141 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     1201 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     1261 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta tggctgatta
     1321 tgatcagtta tctagatccg gtggatctgt cgactgcaga gttcgaagct taagctcgag
     1381 gaccaccagt accaccagca ctaccaccag cactaccacc agcactacca ccagcactac
     1441 caccagcact ggaaagggct ttcttcaagt gcaccttgac cttggttggt cccctgcttg
     1501 tcggtatcgt ccattcgaac atggtgttca gattccctga cgctgcgcct tccttgaatt
     1561 tgccgtaaaa ctcgggagta tccgttccag gtgcaacatc cttgaagaag ttcttcccaa
     1621 tcacctgttt gggatctctg ccagtgatgt ccccttcagc agcattgtac agcaggatat
     1681 tcccgtcacc atcgagctga attacgccaa aggccaacct atccagttgt tcgtcgtcca
     1741 tattggccag agtgttctcg atgtcctcac tgccaaaggc aacatgctcc attccacttg
     1801 ctcccttgta cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca
     1861 ggaccatgtg atcgcgcttc tcgttggggt ctttgctcag ggcggactgg gtgctcaggt
     1921 agtggttgtc gggcagcagc acggggccgt cgccgatggg ggtgttctgc tggtagtggt
     1981 cggcgagctg cacgctgccg tcctcgatgt tgtggcggat cttgaagttg gccttgatgc
     2041 cgttcttctg cttgtcggcg gtgatataga cgttgtggct gatgtagttg tactccagct
     2101 tgtgccccag gatgttgccg tcctccttga agtcgatgcc cttcagctcg atgcggttca
     2161 ccagggtgtc gccctcgaac ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga
     2221 agaagatggt gcgctcctgg acgtagcctt cgggcatggc ggacttgaag aagtcgtgct
     2281 gcttcatgtg gtcggggtag cggctgaagc actgcacgcc ccaggtcagg gtggtcacga
     2341 gggtgggcca gggcacgggc agcttgccgg tggtgcagat gaacttcagg gtcagcttgc
     2401 cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc
     2461 cgtccagctc gaccaggatg ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca
     2521 taccaccagc actaccacca gcactggatc cgaagttggg gtcacttcgt cttttgcggt
     2581 caaagtagat gcagtaccct atgaagaggg caccgcacac gcccgcggcg atggcgctgt
     2641 tccgacccac catggtggcg aattcaccgg tagcgctagc ggatctgacg gttcactaaa
     2701 ccagctctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg
     2761 gcggagttgt tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca
     2821 ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca
     2881 ttgatgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac
     2941 tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta
     3001 ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag
     3061 tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt
     3121 actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca
     3181 ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat
     3241 gaccccgtaa ttgattacta ttaataacta atgcatggcg gtaatacggt tatccacaga
     3301 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3361 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3421 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3481 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3541 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3601 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3661 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3721 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtga
     3781 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3841 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3901 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3961 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4021 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4081 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaac ctgaggctat
     4141 ggcagggcct gccgccccga cgttggctgc gagccctggg ccttcacccg aacttggggg
     4201 gtggggtggg gaaaaggaag aaacgcgggc gtattggccc caatggggtc tcggtggggt
     4261 atcgacagag tgccagccct gggaccgaac cccgcgttta tgaacaaacg acccaacacc
     4321 gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg gttccttccg gtattgtctc
     4381 cttccgtgtt tcagttagcc tccccctagg gtgggcgaag aactccagca tgagatcccc
     4441 gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata
     4501 gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat
     4561 ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc
     4621 tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca
     4681 agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc
     4741 agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag
     4801 caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg
     4861 gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca
     4921 agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat
     4981 gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact
     5041 ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc
     5101 agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc
     5161 gtggccagcc acgatagccg cgctgcctcg tcttgcagtt cattcagggc accggacagg
     5221 tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca
     5281 gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc
     5341 ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct
     5401 tgatcgatct ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag
     5461 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
     5521 cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac
     5581 tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg ggg
//
