LOCUS       sequence-406795-        6381 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses p65-mCherry-FIREtag in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6381)
  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 6381)
  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: pAG1244 p65-mCherry-FIREtag
FEATURES             Location/Qualifiers
     source          1..6381
                     /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             1896..2267
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALLSQISS"
     CDS             2283..2990
                     /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(2428..2446)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2696..2715
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3169..3290
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3206..3225)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3260..3279
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3297..3752)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3434..3455)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3646..3665
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3779..3883
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3885..4242
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3885..4076
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3908..3928)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4093..4228
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4277..5071
                     /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             4277..5071
                     /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(4331..4350)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4941..4960
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5259..5278)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5303..5350
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5679..6267
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6168..6187
                     /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 atctatggac gaactgttcc ccctcatctt cccggcagag ccagcccagg
      661 cctctggccc ctatgtggag atcattgagc agcccaagca gcggggcatg cgcttccgct
      721 acaagtgcga ggggcgctcc gcgggcagca tcccaggcga gaggagcaca gataccacca
      781 agacccaccc caccatcaag atcaatggct acacaggacc agggacagtg cgcatctccc
      841 tggtcaccaa ggaccctcct caccggcctc acccccacga gcttgtagga aaggactgcc
      901 gggatggctt ctatgaggct gagctctgcc cggaccgctg catccacagt ttccagaacc
      961 tgggaatcca gtgtgtgaag aagcgggacc tggagcaggc tatcagtcag cgcatccaga
     1021 ccaacaacaa ccccttccaa gttcctatag aagagcagcg tggggactac gacctgaatg
     1081 ctgtgcggct ctgcttccag gtgacagtgc gggacccatc aggcaggccc ctccgcctgc
     1141 cgcctgtcct ttctcatccc atctttgaca atcgtgcccc caacactgcc gagctcaaga
     1201 tctgccgagt gaaccgaaac tctggcagct gcctcggtgg ggatgagatc ttcctactgt
     1261 gtgacaaggt gcagaaagag gacattgagg tgtatttcac gggaccaggc tgggaggccc
     1321 gaggctcctt ttcgcaagct gatgtgcacc gacaagtggc cattgtgttc cggacccctc
     1381 cctacgcaga ccccagcctg caggctcctg tgcgtgtctc catgcagctg cggcggcctt
     1441 ccgaccggga gctcagtgag cccatggaat tccagtacct gccagataca gacgatcgtc
     1501 accggattga ggagaaacgt aaaaggacat atgagacctt caagagcatc atgaagaaga
     1561 gtcctttcag cggacccacc gacccccggc ctccacctcg acgcattgct gtgccttccc
     1621 gcagctcagc ttctgtcccc aagccagcac cccagcccta tccctttacg tcatccctga
     1681 gcaccatcaa ctatgatgag tttcccacca tggtgtttcc ttctgggcag atcagccagg
     1741 cctcggcctt ggccccggcc cctccccaag tcctgcccca ggctccagcc cctgcccctg
     1801 ctccagccat ggtatcagct ctggcccagg ccccagcccc tgtcccagtc ctagccccag
     1861 gccctcctca ggctgtggcc ccacctgccc ccaagcccac ccaggctggg gaaggaacgc
     1921 tgtcagaggc cctgctgcag ctgcagtttg atgatgaaga cctgggggcc ttgcttggca
     1981 acagcacaga cccagctgtg ttcacagacc tggcatccgt cgacaactcc gagtttcagc
     2041 agctgctgaa ccagggcata cctgtggccc cccacacaac tgagcccatg ctgatggagt
     2101 accctgaggc tataactcgc ctagtgacag gggcccagag gccccccgac ccagctcctg
     2161 ctccactggg ggccccgggg ctccccaatg gcctcctttc aggagatgaa gacttctcct
     2221 ccattgcgga catggacttc tcagccctgc tgagtcagat cagctcctcc ggaggaggcg
     2281 gcatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
     2341 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     2401 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
     2461 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
     2521 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
     2581 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
     2641 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     2701 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     2761 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     2821 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     2881 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     2941 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag tctagaggcg
     3001 gcggctccgg tgacagatat tgggtctttg tgaaacgggt gtaatgagcg gccgcgactc
     3061 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     3121 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     3181 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     3241 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat
     3301 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     3361 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     3421 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     3481 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     3541 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     3601 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     3661 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3721 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     3781 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3841 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     3901 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3961 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4021 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4081 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4141 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4201 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     4261 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4321 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4381 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4441 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4501 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4561 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4621 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4681 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4741 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4801 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4861 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4921 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4981 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5041 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     5101 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5161 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5221 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     5281 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     5341 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     5401 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     5461 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     5521 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5581 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5641 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5701 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5761 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5821 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5881 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5941 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6001 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6061 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6121 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6181 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6241 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6301 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6361 aaccgtatta ccgccatgca t
//
