LOCUS       sequence-406793-        5244 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FIREmate-ECFP-Fis1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5244)
  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 5244)
  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: pAG1237 FIREmate-ECFP-Fis1
FEATURES             Location/Qualifiers
     source          1..5244
                     /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             join(984..986,987..989,990..1700)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1029..1050)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1290..1309)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1637..1658
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2032..2153
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2069..2088)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2123..2142
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2160..2615)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2297..2318)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2509..2528
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2642..2746
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2748..3105
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2748..2939
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2771..2791)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2956..3091
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3140..3934
                     /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             3140..3934
                     /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(3194..3213)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3804..3823
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4122..4141)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4166..4213
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4542..5130
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5031..5050
                     /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 gctagcgcta
      601 ccggactcag atctatgatg gagcatgttg cctttggcag tgaggacatc gagaacactc
      661 tggccaatat ggacgacgaa caactggata ggttggcctt tggcgtaatt cagctcgatg
      721 gtgacgggaa tatcctgctg tacaatgctg ctgaagggga catcactggc agagatccca
      781 aacaggtgat tgggaagaac ttcttcaagg atgttgcacc tggaacggat actcccgagt
      841 tttacggcaa attcaaggaa ggcgcagcgt cagggaatct gaacaccatg ttcgaatgga
      901 cgataccgac aagcagggga ccaaccaagg tcaaggtgca cttgaagaaa gccctttccg
      961 gtggtgctag tggtggtgct agtatggtga gcaagggcga ggagctgttc accggggtgg
     1021 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1081 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     1141 agctgcccgt gccctggccc accctcgtga ccaccctgac ctggggcgtg cagtgcttca
     1201 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1261 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1321 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1381 aggacggcaa catcctgggg cacaagctgg agtacaacta catcagccac aacgtctata
     1441 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg
     1501 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1561 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     1621 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1681 gcatggacga gctgtacaag tgtacaagtc tagaggcggc ggcttacgtc cgcgggttgc
     1741 tgcagacaga gccccagaac aaccaggcca aggaactgga gcggctcatt gacaaggcca
     1801 tgaagaaaga tggactcgtg ggcatggcca tcgtgggagg catggccctg ggtgtggcgg
     1861 gactggccgg actcatcgga cttgctgtgt ccaagtccaa atcctaatga gcggccgcga
     1921 ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     1981 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     2041 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     2101 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta
     2161 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     2221 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     2281 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     2341 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     2401 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     2461 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     2521 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     2581 acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt
     2641 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2701 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag
     2761 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2821 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2881 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     2941 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3001 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3061 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac
     3121 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     3181 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     3241 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     3301 cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg
     3361 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     3421 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     3481 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     3541 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     3601 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     3661 caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     3721 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     3781 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     3841 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     3901 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     3961 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     4021 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     4081 ctcatgctgg agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca
     4141 ataccggaag gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt
     4201 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     4261 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     4321 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc
     4381 tcaggttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4441 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4501 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4561 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4621 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4681 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4741 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4801 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4861 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4921 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4981 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5041 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5101 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5161 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5221 gataaccgta ttaccgccat gcat
//
