LOCUS       sequence-407150-        7880 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 7880)
  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 7880)
  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-mCherry-Bruno isoform A (JM78)
FEATURES             Location/Qualifiers
     source          1..7880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2565
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             2582..3289
                     /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(2727..2745)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2995..3014
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             5186..5227
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5237..5254
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5277..5294)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5327..5461
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5351..5370)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5405..5424
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6062..6650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6142..6161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6821..7612,7613..7681))
                     /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     7444..7463
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7682..7786)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7854..7872
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cttcaagaat taattctata ttctaaaaac acaaatgata cttctaaaaa aaaatcatga
       61 atggcatcaa ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac
      121 atcatttttc cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg
      181 ccattaaagc tggctgatcg tcgccaaaca ccaaatacat aatgaatatg tacacattcg
      241 agaaagaagc gatcaaagaa gcgtcttcgg gcggagtagg agaatgcgga ggagaaggag
      301 aacgagctga tctagtatct ctccacaatc caatgccaac tgaccaactg gccatattcg
      361 gagcaatttg aagccaattt ccatcgcctg gcgatcgctc cattcttggc tatatgtttt
      421 tcaccgttac ccggggccat tttcaaagac tcgtcggcaa gataagattg tgtcactcgc
      481 tgtctctctt catttgtcga agaatgctga ggaatttcgc gatgacgtcg gcgagtattt
      541 tgaagaatga gaataatttg tatttatacg aaaatcagtt agtggaattt tctacaaaaa
      601 catgttatct atagataatt ttgttgcaaa atatgttgac tatgacaaag attgtatgta
      661 tataccttta atgtattctc attttcttat gtatttataa tggcaatgat gatactgatg
      721 atattttaag atgatgccag accaaaaggc ttgaatttct gcgtcttttg ccgaacgcag
      781 tgcatgtgca attgttgttt tttggaatat tcaattttcg gactgtccgc tttgatttca
      841 gtttcttggc ttattcaaaa agcaaagtaa agccaaaaaa gcgagatggc aataccaaat
      901 gcggcaaaac ggtagtggaa ggaaaggggt gcggggcagc ggaaggaagg gtggggcggg
      961 gcgtggcggg gtctgtggct gggcgcgacg tcaccgacgt tggagccact cctttgacca
     1021 tgtgtgcgtg tgtgtattat tcgtgtctcg ccactcgccg gttgtttttt tctttttatg
     1081 ctgcgctctc tctagcgcca tctcgcttac gcatgctcaa cgcaccgcat gttgccgttt
     1141 ccttttatgc gtcattttgg ctcgaaatag gcaattattt aaacaaagat tagtcaacga
     1201 aaacgctaaa ataaataagt ctacaatatg gttacttatt gccatgtgtg tgcagccaac
     1261 gatagcaaca aaagcaacaa cacaggtggc tttccctctt tcactttttg tttgcaagcc
     1321 gcgtgcgagc aagacggcac gaccggcaaa cgcaattacg ctgacaaaga gcagacgaag
     1381 ttttggcgaa aaacatcaag gcgcctgata cgaatgcatt tgcaataaca attgcgatat
     1441 ttaatattgt ttatgaagct gtttgacttc aaaacacaca aaaaaaaaaa taaaacaaat
     1501 tatttgaaag agaattagga atcggacgct tatcgttagg gtaacaacaa gaaatgctta
     1561 ctgagtcaca gcctctggaa aactgccgca agccagagag agagagaaaa agagggagag
     1621 cagcttagac cgcatgtgct tgtgtgtgag gcgtctctct cttcgtctct gttgcgcaaa
     1681 cgcatagact gcactgaaaa aatcgattac ctatttttta tgaatgaata tttgcactat
     1741 tactattcaa aactattaag atagcaatca cattcaatag ccaaatacta taccacctga
     1801 gcgatgcaac gaaatgatca atttgagcaa aaatgctgca tatttaggac ggcatcatta
     1861 tagaaatgct tcttgctgtg tacttttctc tcgtctggca gctgtttcgc cgttattgtt
     1921 aaaaccggct taagttaggt gtgttttcta cgactagtga atgccctact agaagatgtg
     1981 tgttgcacaa aatgtccctg gaataaccaa tttgaagtgc agatagcagt aaacgtaagc
     2041 taatatgaat attatttaac tgtaatgttt taatatcgct ggacattact aataaaccca
     2101 ctataaacac atgtacatat gtatgttttg gcatacaatg agtagttggg gaaaaaatgt
     2161 gtaaaagcac cgtgaccatc acagcataaa gataaccagc tgaagtatcg aatatgagta
     2221 acccccaaat tgaatcacat gccgcaactg ataggaccca tggaagtaca ctcttcatgg
     2281 cgatatacaa gacacacaca agcacgaaca cccagttgcg gaggaaattc tccgtaaatg
     2341 aaaacccaat cggcgaacaa ttcataccca tatatggtaa aagttttgaa cgcgacttga
     2401 gagcggagag cattgcggct gataaggttt tagcgctaag cgggctttat aaaacgggct
     2461 gcgggaccag ttttcatatc actaccgttt gagttcttgt gctgtgtgga tactcctccc
     2521 gacacaaagc cgctccatca gccagcagtc gtctaatcca gagaccccgg atcggggtac
     2581 catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
     2641 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     2701 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     2761 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     2821 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     2881 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     2941 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     3001 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     3061 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     3121 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     3181 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     3241 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagc caagcttgaa
     3301 ttctgcaacg tgcatgttca ccagccgcgc ttcgtttttg gccaatcgcc gcatgatttt
     3361 cgatttctcc gagaagaaca acgatatcta cttggatgct tttggtatga gtgataagag
     3421 cagcagtgca acgaacagcc tgccgaatag tcccatacac agcaacaaca ataatccatc
     3481 gctgctgaac aacaacaaca ataacagcaa cggcaccagc agcaacaaca gcttaaacgt
     3541 tggcaacaac aacagcaatc cgagcttggg cggcagcaac agcaacgcac tggtttccgt
     3601 aggcagcaac ggtataatgt ccgccgccgg tttggtcaac aacaataaca atccgtgcag
     3661 cgccaaccgc aatgtggtcg ccatggtcga cgatgacgcc tgcttccgtc tggatacgga
     3721 cgccacggtg acgtacggcg agaaggaacc ggatccggac aacatcaaga tgttcgtcgg
     3781 ccaggtgccc aagtcgatgg acgagtcgca gctgcgcgag atgttcgagg agtacggcgc
     3841 cgtgcactcg atcaacgttc tgcgggacaa ggctaccgga attagcaagg gttgctgctt
     3901 tgtgacattt tatacgagac acgccgccct gaaagcccag gatgctttgc ataatgttaa
     3961 gacgctaaat gggatgtacc atcccattca aatgaagccc gccgatagcg agaatcgcaa
     4021 tgaacgcaaa ctctttgtgg gtatgctaaa caagaagctg aacgagaacg acgttcgcaa
     4081 gctatttgaa gtccacggag ccatcgagga gtgcaccgtc cttcgggacc aaaacggaca
     4141 gagcaagggt tgtgccttcg tcacattcgc caccaagcac gctgccattt cggccattaa
     4201 agtgaccctg agccagaaca agatcatgga gggctgcaca tcgccgctgg tcgtcaagtt
     4261 cgccgatacg caaaaagaga aggagcagaa gaagattcag caaattcagg ccaatctctg
     4321 gaacctggcc agcaacatca acatacctct gggccaaacg gccaccagcg tgaccacgcc
     4381 catcctgccc aatccgccgc agcagccttc gccggttctc ggcgcagatg ccattacccc
     4441 ggcctccatt caactgctgc agcagctgca ggcggtggga ctgcagcacc agttgttgca
     4501 agccctcaca ggactgggag cccagcagag cagctcggcg acggacacca gcgctgtggc
     4561 ggccgcagga cttttgaccc cgatgaccgt ccagaatcta gcggccattg cggccatgac
     4621 gcagccgagt ctctcgaatg ccgccgcggc agccgccgct gccacatcgc ctggaagcgc
     4681 ccaactgacc aacacggctg cgctgctctg gtccgatccg aatccaatgg catcggcata
     4741 tatgtcaaca gccgccggtc tgccgcaatt cgggagcgcc agtgcattgt ccacttcgcc
     4801 gctggccagc gttgccctca gcgccgcagc cgccgccgca gcgggaaagc agattgaagg
     4861 acccgagggc tgcaaccttt tcatctacca tctgccgcag gagttcaccg atacggatct
     4921 ggcctccacg ttcctgccct tcggcaatgt gatctcggcc aaggtgttca tcgacaaaca
     4981 gacgagcctg tcgaagtgct tcgggtttgt ctccttcgac aatccggact cggcccaggt
     5041 ggccataaag gcgatgaacg gcttccaggt gggcaccaag cgtctcaagg tgcagctgaa
     5101 gaagcccaag gactcgaagc cctactaggc atgagatatc cagcacagtg gcggccgctc
     5161 gagtctagag ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc tcggtctcga
     5221 ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct gatcagcctc
     5281 gactgtgcct tctaaggcct gagctcgctg atcagcctcg atcgaggatc cagacatgat
     5341 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     5401 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     5461 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     5521 ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcagt cgaccgatgc
     5581 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg
     5641 ccgcacttat gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct
     5701 gggtcatttt cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg
     5761 cggtattcgg aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac
     5821 gtttcggcga gaagcaggcc attatcgccg gcatggcggc cgacgcgctg ggctacgtct
     5881 tgctggcgtt cgcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg
     5941 gcggcatcgg gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc
     6001 agggacagct tcaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6061 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6121 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6181 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6241 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6301 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6361 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6421 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6481 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     6541 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6601 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6661 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6721 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6781 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6841 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6901 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6961 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7021 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7081 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag
     7141 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7201 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7261 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7321 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7381 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac
     7441 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7501 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7561 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7621 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7681 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7741 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7801 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     7861 gtatcacgag gccctttcgt
//
