LOCUS       sequence-407181-        8294 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bruno expression in Schneider cells for imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8294)
  AUTHORS   Salgania HK, Metz J, Jeske M
  TITLE     ReLo is a simple and rapid colocalization assay to identify and 
            characterize direct protein-protein interactions.
  JOURNAL   Nat Commun. 2024 Apr 3;15(1):2875. doi: 10.1038/s41467-024-47233-4.
  PUBMED    38570497
REFERENCE   2  (bases 1 to 8294)
  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 
            Commun."; date: "2024-04-3"; volume: "15(1):2875. doi"; pages: " 
            10.1038/s41467-024-47233-4"
COMMENT     Sequence Label: pAc5.1-PH-mEGFP-Bruno isoform A (JM165)
FEATURES             Location/Qualifiers
     source          1..8294
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..2517
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2939..2941,2942..2944,2945..3655)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2984..3005)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3245..3264)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3592..3613
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5552..5593
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5603..5620
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5643..5660)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5693..5827
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5717..5736)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5771..5790
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6428..7016)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6508..6527)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7187..7978,7979..8047))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7810..7829
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8048..8152)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8220..8238
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aaaaaatcat gaatggcatc aactctgaat caaatctttg cagatgcacc tacttctcat
       61 ttccactgtc acatcatttt tccagatctc gctgcctgtt atgtggccca caaaccaaga
      121 cacgttttat ggccattaaa gctggctgat cgtcgccaaa caccaaatac ataatgaata
      181 tgtacacatt cgagaaagaa gcgatcaaag aagcgtcttc gggcggagta ggagaatgcg
      241 gaggagaagg agaacgagct gatctagtat ctctccacaa tccaatgcca actgaccaac
      301 tggccatatt cggagcaatt tgaagccaat ttccatcgcc tggcgatcgc tccattcttg
      361 gctatatgtt tttcaccgtt acccggggcc attttcaaag actcgtcggc aagataagat
      421 tgtgtcactc gctgtctctc ttcatttgtc gaagaatgct gaggaatttc gcgatgacgt
      481 cggcgagtat tttgaagaat gagaataatt tgtatttata cgaaaatcag ttagtggaat
      541 tttctacaaa aacatgttat ctatagataa ttttgttgca aaatatgttg actatgacaa
      601 agattgtatg tatatacctt taatgtattc tcattttctt atgtatttat aatggcaatg
      661 atgatactga tgatatttta agatgatgcc agaccaaaag gcttgaattt ctgcgtcttt
      721 tgccgaacgc agtgcatgtg caattgttgt tttttggaat attcaatttt cggactgtcc
      781 gctttgattt cagtttcttg gcttattcaa aaagcaaagt aaagccaaaa aagcgagatg
      841 gcaataccaa atgcggcaaa acggtagtgg aaggaaaggg gtgcggggca gcggaaggaa
      901 gggtggggcg gggcgtggcg gggtctgtgg ctgggcgcga cgtcaccgac gttggagcca
      961 ctcctttgac catgtgtgcg tgtgtgtatt attcgtgtct cgccactcgc cggttgtttt
     1021 tttcttttta tgctgcgctc tctctagcgc catctcgctt acgcatgctc aacgcaccgc
     1081 atgttgccgt ttccttttat gcgtcatttt ggctcgaaat aggcaattat ttaaacaaag
     1141 attagtcaac gaaaacgcta aaataaataa gtctacaata tggttactta ttgccatgtg
     1201 tgtgcagcca acgatagcaa caaaagcaac aacacaggtg gctttccctc tttcactttt
     1261 tgtttgcaag ccgcgtgcga gcaagacggc acgaccggca aacgcaatta cgctgacaaa
     1321 gagcagacga agttttggcg aaaaacatca aggcgcctga tacgaatgca tttgcaataa
     1381 caattgcgat atttaatatt gtttatgaag ctgtttgact tcaaaacaca caaaaaaaaa
     1441 aataaaacaa attatttgaa agagaattag gaatcggacg cttatcgtta gggtaacaac
     1501 aagaaatgct tactgagtca cagcctctgg aaaactgccg caagccagag agagagagaa
     1561 aaagagggag agcagcttag accgcatgtg cttgtgtgtg aggcgtctct ctcttcgtct
     1621 ctgttgcgca aacgcataga ctgcactgaa aaaatcgatt acctattttt tatgaatgaa
     1681 tatttgcact attactattc aaaactatta agatagcaat cacattcaat agccaaatac
     1741 tataccacct gagcgatgca acgaaatgat caatttgagc aaaaatgctg catatttagg
     1801 acggcatcat tatagaaatg cttcttgctg tgtacttttc tctcgtctgg cagctgtttc
     1861 gccgttattg ttaaaaccgg cttaagttag gtgtgttttc tacgactagt gaatgcccta
     1921 ctagaagatg tgtgttgcac aaaatgtccc tggaataacc aatttgaagt gcagatagca
     1981 gtaaacgtaa gctaatatga atattattta actgtaatgt tttaatatcg ctggacatta
     2041 ctaataaacc cactataaac acatgtacat atgtatgttt tggcatacaa tgagtagttg
     2101 gggaaaaaat gtgtaaaagc accgtgacca tcacagcata aagataacca gctgaagtat
     2161 cgaatatgag taacccccaa attgaatcac atgccgcaac tgataggacc catggaagta
     2221 cactcttcat ggcgatatac aagacacaca caagcacgaa cacccagttg cggaggaaat
     2281 tctccgtaaa tgaaaaccca atcggcgaac aattcatacc catatatggt aaaagttttg
     2341 aacgcgactt gagagcggag agcattgcgg ctgataaggt tttagcgcta agcgggcttt
     2401 ataaaacggg ctgcgggacc agttttcata tcactaccgt ttgagttctt gtgctgtgtg
     2461 gatactcctc ccgacacaaa gccgctccat cagccagcag tcgtctaatc cagagacccc
     2521 ggatcggggt accaacatgc acggtctcca ggatgatcca gatctgcagg ctctgctgaa
     2581 aggttctcag ctcctcaaag tcaaaagcag ctcctggcgt cgtgaacgct tctacaaact
     2641 gcaggaggac tgcaaaacca tctggcagga atcccgtaaa gtaatgcgct ccccagagtc
     2701 tcagctcttc agcattgagg acattcagga ggtacgtatg ggtcaccgta ccgaaggtct
     2761 ggagaaattc gctcgcgata tcccagaaga tcgctgcttc tctatcgtct tcaaagacca
     2821 gcgcaacacc ctggatctga ttgctccatc tccagcagat gctcagcact gggttcaggg
     2881 tctccgcaaa atcattcacc actccggttc tatggaccag cgtcagaaag gtaccatcat
     2941 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     3001 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     3061 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     3121 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     3181 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     3241 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     3301 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     3361 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     3421 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     3481 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     3541 cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     3601 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagccaag
     3661 cttgaattct gcaacgtgca tgttcaccag ccgcgcttcg tttttggcca atcgccgcat
     3721 gattttcgat ttctccgaga agaacaacga tatctacttg gatgcttttg gtatgagtga
     3781 taagagcagc agtgcaacga acagcctgcc gaatagtccc atacacagca acaacaataa
     3841 tccatcgctg ctgaacaaca acaacaataa cagcaacggc accagcagca acaacagctt
     3901 aaacgttggc aacaacaaca gcaatccgag cttgggcggc agcaacagca acgcactggt
     3961 ttccgtaggc agcaacggta taatgtccgc cgccggtttg gtcaacaaca ataacaatcc
     4021 gtgcagcgcc aaccgcaatg tggtcgccat ggtcgacgat gacgcctgct tccgtctgga
     4081 tacggacgcc acggtgacgt acggcgagaa ggaaccggat ccggacaaca tcaagatgtt
     4141 cgtcggccag gtgcccaagt cgatggacga gtcgcagctg cgcgagatgt tcgaggagta
     4201 cggcgccgtg cactcgatca acgttctgcg ggacaaggct accggaatta gcaagggttg
     4261 ctgctttgtg acattttata cgagacacgc cgccctgaaa gcccaggatg ctttgcataa
     4321 tgttaagacg ctaaatggga tgtaccatcc cattcaaatg aagcccgccg atagcgagaa
     4381 tcgcaatgaa cgcaaactct ttgtgggtat gctaaacaag aagctgaacg agaacgacgt
     4441 tcgcaagcta tttgaagtcc acggagccat cgaggagtgc accgtccttc gggaccaaaa
     4501 cggacagagc aagggttgtg ccttcgtcac attcgccacc aagcacgctg ccatttcggc
     4561 cattaaagtg accctgagcc agaacaagat catggagggc tgcacatcgc cgctggtcgt
     4621 caagttcgcc gatacgcaaa aagagaagga gcagaagaag attcagcaaa ttcaggccaa
     4681 tctctggaac ctggccagca acatcaacat acctctgggc caaacggcca ccagcgtgac
     4741 cacgcccatc ctgcccaatc cgccgcagca gccttcgccg gttctcggcg cagatgccat
     4801 taccccggcc tccattcaac tgctgcagca gctgcaggcg gtgggactgc agcaccagtt
     4861 gttgcaagcc ctcacaggac tgggagccca gcagagcagc tcggcgacgg acaccagcgc
     4921 tgtggcggcc gcaggacttt tgaccccgat gaccgtccag aatctagcgg ccattgcggc
     4981 catgacgcag ccgagtctct cgaatgccgc cgcggcagcc gccgctgcca catcgcctgg
     5041 aagcgcccaa ctgaccaaca cggctgcgct gctctggtcc gatccgaatc caatggcatc
     5101 ggcatatatg tcaacagccg ccggtctgcc gcaattcggg agcgccagtg cattgtccac
     5161 ttcgccgctg gccagcgttg ccctcagcgc cgcagccgcc gccgcagcgg gaaagcagat
     5221 tgaaggaccc gagggctgca accttttcat ctaccatctg ccgcaggagt tcaccgatac
     5281 ggatctggcc tccacgttcc tgcccttcgg caatgtgatc tcggccaagg tgttcatcga
     5341 caaacagacg agcctgtcga agtgcttcgg gtttgtctcc ttcgacaatc cggactcggc
     5401 ccaggtggcc ataaaggcga tgaacggctt ccaggtgggc accaagcgtc tcaaggtgca
     5461 gctgaagaag cccaaggact cgaagcccta ctaggcatga gatatccagc acagtggcgg
     5521 ccgctcgagt ctagagggcc cgcggttcga aggtaagcct atccctaacc ctctcctcgg
     5581 tctcgattct acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc
     5641 agcctcgact gtgccttcta aggcctgagc tcgctgatca gcctcgatcg aggatccaga
     5701 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     5761 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     5821 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     5881 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg atcagtcgac
     5941 cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta
     6001 tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag
     6061 cgctctgggt cattttcggc gaggaccgct ttcgctggag cgcgacgatg atcggcctgt
     6121 cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca
     6181 ccaaacgttt cggcgagaag caggccatta tcgccggcat ggcggccgac gcgctgggct
     6241 acgtcttgct ggcgttcgcg acgcgaggct ggatggcctt ccccattatg attcttctcg
     6301 cttccggcgg catcgggatg cccgcgttgc aggccatgct gtccaggcag gtagatgacg
     6361 accatcaggg acagcttcaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6421 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6481 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6541 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6601 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6661 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6721 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6781 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6841 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     6901 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6961 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7021 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7081 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7141 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7201 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7261 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7321 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7381 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7441 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7501 ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7561 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7621 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7681 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7741 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7801 caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7861 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7921 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7981 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8041 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8101 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8161 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     8221 ataggcgtat cacgaggccc tttcgtcttc aagaattaat tctatattct aaaaacacaa
     8281 atgatacttc taaa
//
