LOCUS       sequence-406791-        5528 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses H2B-ECFP-FIREmate in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5528)
  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 5528)
  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: pAG1247 H2B-ECFP-FIREmate
FEATURES             Location/Qualifiers
     source          1..5528
                     /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(1014..1016,1017..1019,1020..1730)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1059..1080)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1320..1339)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1667..1688
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2316..2437
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2353..2372)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2407..2426
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2444..2899)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2581..2602)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2793..2812
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2926..3030
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3032..3389
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3032..3223
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3055..3075)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3240..3375
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3424..4218
                     /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             3424..4218
                     /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(3478..3497)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4088..4107
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4406..4425)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4450..4497
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4826..5414
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5315..5334
                     /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 ccggtgaatt cgccaccatg atgcccgaac ctgcgaagtc agcgcccgct cccaaaaaag
      661 gctctaaaaa agctgtcgcc aagacccaga agaaggggga taagaaaagg cgtaagacca
      721 ggaaagagag ttacgccatt tacgtgtaca aagtactaaa acaagtccac ccggacactg
      781 gcatctcctc aaaggcgatg ggcattatga actcatttgt aaacgacatc ttcgagcgca
      841 tcgccggaga agcgtcgcgc ctggcgcatt acaacaagcg ctccactatc acatcccggg
      901 agatccagac ggccgtgcgc ctgctcctgc ccggagaact ggccaaacac gctgtgtctg
      961 agggcacaaa ggccgtgacc aagtacacca gctccaaggg tagtgctggt ggtatggtga
     1021 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     1081 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     1141 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     1201 ccaccctgac ctggggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
     1261 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     1321 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     1381 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     1441 agtacaacta catcagccac aacgtctata tcaccgccga caagcagaag aacggcatca
     1501 aggccaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     1561 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1621 gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1681 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag ggagcaagtg
     1741 gaatggagca tgttgccttt ggcagtgagg acatcgagaa cactctggcc aatatggacg
     1801 acgaacaact ggataggttg gcctttggcg taattcagct cgatggtgac gggaatatcc
     1861 tgctgtacaa tgctgctgaa ggggacatca ctggcagaga tcccaaacag gtgattggga
     1921 agaacttctt caaggatgtt gcacctggaa cggatactcc cgagttttac ggcaaattca
     1981 aggaaggcgc agcgtcaggg aatctgaaca ccatgttcga atggacgata ccgacaagca
     2041 ggggaccaac caaggtcaag gtgcacttga agaaagccct ttccagtgct ggtggtagtg
     2101 ctggtggtag tgctggtggt agtgctggtg gtagtgctgg tggtactggt ggtcctcgag
     2161 cttaagcttc gaactctgca gtcgacagat ccaccggatc tagataactg atcataatca
     2221 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2281 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2341 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     2401 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     2461 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     2521 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     2581 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     2641 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     2701 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     2761 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     2821 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     2881 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2941 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3001 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     3061 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3121 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3181 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3241 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3301 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3361 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     3421 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3481 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3541 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3601 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3661 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3721 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3781 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3841 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3901 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     3961 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4021 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4081 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4141 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4201 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4261 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4321 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4381 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     4441 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     4501 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     4561 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     4621 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     4681 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4741 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4801 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4861 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4921 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4981 tgtagccgta gttaggccac cacttcaaga actctgtatc accgcctaca tacctcgctc
     5041 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5101 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5161 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5221 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5281 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5341 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5401 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5461 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     5521 ccatgcat
//
