LOCUS       sequence-320526-        7222 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human BRAT1 with a N-terminal GFP-FLAG tag. Confers G418 
            resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7222)
  AUTHORS   Demange P, Joly E, Marcoux J, Zanon PRA, Listunov D, Rulliere P, 
            Barthes C, Noirot C, Izquierdo JB, Rozie A, Pradines K, Hee R, de 
            Brito MV, Marcellin M, Serre RF, Bouchez O, Burlet-Schiltz O, 
            Oliveira MCF, Ballereau S, Bernardes-Genisson V, Maraval V, Calsou 
            P, Hacker SM, Genisson Y, Chauvin R, Britton S
  TITLE     SDR enzymes oxidize specific lipidic alkynylcarbinols into cytotoxic
            protein-reactive species.
  JOURNAL   Elife. 2022 May 10;11. pii: 73913. doi: 10.7554/eLife.73913.
  PUBMED    35535493
REFERENCE   2  (bases 1 to 7222)
  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: "Elife."; 
            date: "2022-05-10"; volume: "11. pii: 73913. doi"; pages: " 
            10.7554/eLife.73913"
COMMENT     Sequence Label: pEGFP-C1-FLAG-BRAT1
FEATURES             Location/Qualifiers
     source          1..7222
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..3,4..6,7..717)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1..3,4..6,7..717)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(46..67)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(307..326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     654..675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             733..756
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3398..3519
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3435..3454)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3489..3508
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3526..3981)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3663..3684)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3875..3894
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4008..4112
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4114..4471
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4114..4305
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4137..4157)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4322..4457
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4506..5300
                     /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             4506..5300
                     /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(4560..4579)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5170..5189
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5488..5507)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5532..5579
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5908..6496
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6397..6416
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6671..6974
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6975..7178
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7129..7149
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7217..7222,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
       61 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      121 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      181 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
      241 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      301 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      361 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      421 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
      481 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
      541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      601 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      661 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtcc
      721 ggactcagat ctgattacaa ggatgacgac gataagctcg agggcggccg catggaccca
      781 gaatgcgccc agctgctccc ggctctctgt gctgttctgg tagatcccgg gcagccggtg
      841 gcagatgaca cctgtttgga gaagctcctg gactggttta aaacggtcac tgaaggagag
      901 tccagtgtcg tgctgctgca ggagcacccc tgcctggtgg agctgctgtc ccatgtgctg
      961 aaagtccagg acctgagttc tggggtcctc tccttctcac tgcgcctggc aggaaccttc
     1021 gcagcccagg aaaactgctt ccagtatctt cagcaggggg agttactacc agggctcttt
     1081 ggggagccag gacccctcgg ccgagcaacc tgggccgtcc ccaccgtgcg cagcggctgg
     1141 atccagggcc tgcgctccct ggcacagcac cccagcgccc tgcgcttcct ggccgaccat
     1201 ggtgcggtcg acaccatctt ctccctgcag ggagactcca gcctgtttgt ggcctcggcg
     1261 gccagtcagc tcctggtgca cgtcctggct ttgtccatgc gaggtggagc cgaggggcag
     1321 ccctgcctgc cggggggtga ctggcccgcg tgtgcccaga agatcatgga tcacgttgaa
     1381 gagtccttgt gctccgcggc cacccccaag gtcactcagg ccctgaacgt cctgaccacg
     1441 accttcgggc gctgccagag cccctggacg gaagccctgt gggtgcggct gagtccccgc
     1501 gtggcctgtc tgctggagag agaccccatc cccgccgcac actcgttcgt ggacctgctt
     1561 ctctgtgtgg ctcgttctcc cgtgttcagt tcttccgacg gcagcctgtg ggagacagtg
     1621 gcgcgggctc tgagctgcct gggtcccacc cacatgggac ccctggcttt ggggatcctg
     1681 aagctcgagc actgtccaca ggcactgagg acccaggcct tccaggtcct tctccagccc
     1741 ctggcctgtg tcctgaaggc cacggttcag gcccccggac ccccaggctt gctggacggg
     1801 acggcagacg atgccacgac ggtggacaca ctcctggcct ccaagtcgtc ctgcgccggc
     1861 ctcctgtgcc gcaccctggc tcacctggag gagctgcagc cgctgcccca gcgcccttca
     1921 ccgtggcccc aggcgtctct actgggggct acagtgactg tcctgcggct ctgtgacggc
     1981 tcggctgccc ctgcctccag tgtggggggc cacctctgtg ggaccctggc gggctgcgtc
     2041 cgggtccagc gagcagccct cgacttcctg gggacgctgt cacaggggac aggcccccag
     2101 gagctggtga cgcaggcgct tgctgtcctc ctggagtgcc tcgagagccc cggctccagc
     2161 cccacggttc tgaagaaggc cttccaggcc acgctcaggt ggctcctgag ctcacccaag
     2221 acccccggct gctctgatct cggccccctc atcccgcagt tcctcagaga gctgttccct
     2281 gtgctgcaga aacgcctgtg ccacccctgc tgggaggtga gggactccgc cctcgagttc
     2341 ctgacccagc tgagcaggca ctggggagga caggctgact tcagatgcgc actcttggct
     2401 tcagaggtgc ctcagctggc cctgcagctc ctccaggacc ctgagagtta tgtccgagcg
     2461 agtgcagtga ccgccatggg gcagctgtcc agccagggcc tgcacgcccc caccagccct
     2521 gagcatgcag aggcccggca gagcctgttc ctggagctcc tgcacatcct ctccgtagac
     2581 tcggagggct tcccacggcg ggcggtcatg caagtcttca ctgagtggct gcgggacggc
     2641 cacgccgacg cggcccagga cacggagcag ttcgtggcca ctgtgctgca ggcggcgagc
     2701 cgagacctgg actgggaggt ccgcgcccag ggcctggagc tggccctcgt gttcctgggc
     2761 cagactttgg ggccgccgcg tacccactgc ccctatgccg tggccctacc cgaggtggcc
     2821 ccagcccagc cactcaccga ggcactgagg gctctctgcc acgtggggct ctttgacttc
     2881 gccttttgtg ccttgtttga ctgcgaccgc cctgtggcgc agaagtcttg tgacctcctt
     2941 ctcttcctga gggacaagat tgcttcctac agcagcctgc gggaggccag gggcagcccc
     3001 aacactgcct ccgcagaggc caccctgccg aggtggcggg cgggtgagca ggcccagccc
     3061 ccaggggacc aggagcctga ggctgtgctg gccatgctca ggtccctaga cctggagggc
     3121 ctgcggagca cgctggccga gagcagcgac cacgtggaaa agagtcccca gtccctcctg
     3181 caggacatgc tggccacggg aggcttcctg cagggggacg aggccgactg ctactgagaa
     3241 ttctgcagtc gacggtaccg cgggcccggg atccaccgga tctagataac tgatcataat
     3301 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     3361 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     3421 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     3481 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta
     3541 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     3601 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     3661 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     3721 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     3781 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     3841 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     3901 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     3961 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4021 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4081 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     4141 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4201 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4261 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4321 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     4381 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4441 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     4501 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     4561 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     4621 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     4681 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     4741 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     4801 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     4861 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     4921 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     4981 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     5041 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5101 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5161 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5221 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     5281 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     5341 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5401 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5461 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     5521 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     5581 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     5641 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     5701 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     5761 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5821 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5881 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5941 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6001 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6061 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6121 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6181 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6241 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6301 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6361 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6421 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6481 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6541 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6601 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     6661 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     6721 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     6781 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     6841 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     6901 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     6961 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     7021 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     7081 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     7141 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc
     7201 gctagcgcta ccggtcgcca cc
//
