LOCUS       sequence-401683-        8690 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Aubergine expression in Schneider cells for imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8690)
  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 8690)
  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-EGFP-Aubergine 4R-K (R11K/R13K/R15K/R17...
FEATURES             Location/Qualifiers
     source          1..8690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..2520
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2540..2542,2543..2545,2546..3256)
                     /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(2585..2606)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2846..2865)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3193..3214
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(3426..3443)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5927..5968
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5978..5995
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6018..6035)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6068..6202
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6092..6111)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6146..6165
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6824..7412)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6904..6923)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7583..8374,8375..8443))
                     /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     8206..8225
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8444..8548)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8616..8634
                     /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 atatcaatat
      181 gtacattcga gaaagaagcg atcaaagaag cgtcttcggg cgagtaggag aatgcggagg
      241 agaaggagaa cgagctgatc tagtatctct ccacaatcca atgccaactg accaactggc
      301 catattcgga gcaatttgaa gccaatttcc atcgcctggc gatcgctcca ttcttggcta
      361 tatgtttttc accgttcccg gggccatttt caaagactcg tcggtaagat aagattgtgt
      421 cactcgctgt ctctcttcat ttgtcgaaga atgctgagga atttcgcgat gacgtcggcg
      481 agtattttga agaatgagaa taatttgtat ttatacgaaa atcagttagt ggaattttct
      541 acaaaaacat gttatctata gataattttg ttgcaaaata tgttgactat gacaaagatt
      601 gtatgtatat acctttaatg tattctcatt ttcttatgta tttataatgg caatgatgat
      661 actgatgata ttttaagatg atgccagacc acaggctgat ttctgcgtct tttgccgaac
      721 gcagtgcatg tgcggttgtt gttttttgga atagtttcaa ttttcggact gtccgctttg
      781 atttcagttt cttggcttat tcaaaaagca aagtaaagcc aaaaaagcga gatggcaata
      841 ccaaatgcgg caaaacggta gtggaaggaa aggggtgcgg ggcagcggaa ggaagggtgg
      901 ggcggggcgt ggcggggtct gtggctgggc gcgacgtcac cgacgttgga gccactcctt
      961 tgaccatgtg tgcgtgtgtg tattattcgt gtctcgccac tcgccggttg tttttttctt
     1021 tttatctcgc tctctctagc gccatctcgt acgcatgctc aacgcaccgc atgttgccgt
     1081 gtcctttatg cgtcattttg gctcgaaata ggcaattatt taaacaaaga ttagtcaacg
     1141 aaaacgctaa aataaataag tctacaatat ggttacttat tgccatgtgt gtgcagccaa
     1201 cgatagcaac aaaagcaaca acacagtggc tttccctctt tcactttttg tttgcaagcg
     1261 cgtgcgagca agacggcacg accggcaaac gcaattacgc tgacaaagag cagacgaagt
     1321 tttggccgaa aaacatcaag gcgcctgata cgaatgcatt tgcaataaca attgcgatat
     1381 ttaatattgt ttatgaagct gtttgacttc aaaacacaca aaaaaaaaaa taaaacaaat
     1441 tatttgaaag agaattagga atcggacagc ttatcgttac gggctaacag cacaccgaga
     1501 cgaaatagct tacctgacgt cacagcctct ggaagaactg ccgccaagca gagagagaga
     1561 gaaaaagagg gagagcagct tagaccgcat gtgcttgtgt gtgaggcgtc tctctcttcg
     1621 tctcctgttt gcgcaaacgc atagactgca ctgagaaaat cgattaccta ttttttatga
     1681 atgaatattt gcactattac tattcaaaac tattaagata gcaatcacat tcaatagcca
     1741 aatactatac cacctgagcg atgcaacgaa atgatcaatt tgagcaaaaa tgctgcatat
     1801 ttaggacggc atcattatag aaatgcttct tgctgtgtac ttttctctcg tctggcagct
     1861 gtttcgccgt tattgttaaa accggcttaa gttaggtgtg ttttctacga ctagtgatgc
     1921 ccctactaga agatgtgtgt tgcacaaatg tccctgaata accaatttga agtgcagata
     1981 gcagtaaacg taagctaata tgaatattat ttaactgtaa tgttttaata tcgctggaca
     2041 ttactaataa acccactata aacacatgta catatgtatg ttttggcata caatgagtag
     2101 ttggggaaaa aatgtgtaaa agcaccgtga ccatcacagc ataaagataa ccagctgaag
     2161 tatcgaatat gagtaacccc caaattgaat cacatgccgc aactgatagg acccatggaa
     2221 gtacactctt catggcgata tacaagacac acacaagcac gaacacccag ttgcggagga
     2281 aattctccgt aaatgaaaac ccaatcggcg aacaattcat acccatatat ggtaaaagtt
     2341 ttgaacgcga cttgagagcg gagagcattg cggctgataa ggttttagcg ctaagcgggc
     2401 tttataaaac gggctgcggg accagttttc atatcactac cgtttgagtt cttgtgctgt
     2461 gtggatactc ctcccgacac aaagccgctc catcagccag cagtcgtcta atccagagac
     2521 cccggatcgg ggtaccaaca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     2581 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     2641 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     2701 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     2761 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     2821 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     2881 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2941 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     3001 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     3061 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     3121 gctgctgccc gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga
     3181 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     3241 ggacgagctg tacaagccaa gcttgaattc tgcagatatg aatttaccac caaaccctgt
     3301 aattgctaag ggaaagggta agggaaagaa gcccaataat gtagaggcca atcgcggctt
     3361 cgctccttcg ctgggtcaga agtcggaccc ttcccactcg gaaggaaatc aagcatccgg
     3421 aggcaatggt ggtggtggtg atgctcaagt gggtccctcg atagagaaat ccagtttgag
     3481 cgcggtgcaa atgcacaagt ccgagggcga tccgcgtggc tccgtgcgcg gccgtaggct
     3541 aataacggat ttggtgtatt cacgtcctcc agggatgacc agcaagaaag gagtggttgg
     3601 cactcatatt accgtgcagg caaactattt taaggtatta aagcgtccaa actggaccat
     3661 ctaccagtac cgcgtcgatt ttacgcctga tgtggaggct acacgactgc gacggtcttt
     3721 cttgtatgaa cataaaggga tcctgggcgg ctacatcttt gacggaacca atatgttttg
     3781 catcaatcag ttcaaagctg ttcaagatag cccctatgtt ttggaactgg ttacgaagag
     3841 tcgtgctggc gaaaacattg aaattaagat caaggctgtt ggatctgtgc aatctacgga
     3901 tgccgagcag tttcaggtcc ttaatctcat actgcgcagg gccatggagg gcttagactt
     3961 gaagctggtc tcgcgctact actacgatcc tcaagctaag attaatttgg agaatttccg
     4021 catgcaatta tggcctggct atcagacttc gattcgccag cacgaaaatg atatattact
     4081 ttgttccgag atatgccaca aggttatgcg aactgagacc ttgtacaata ttttgtccga
     4141 tgctattcgc gacagtgacg attatcaaag tacatttaag cgtgcagtaa tgggtatggt
     4201 aatactaacg gattataata acaaaaccta tcgcattgac gatgtcgact ttcaatcgac
     4261 gccattgtgc aaatttaaaa ccaatgacgg tgagatttcg tacgtggatt actataagaa
     4321 gcgatacaac ataatcattc gcgacttgaa gcaacctctg gtcatgtctc gtccgacaga
     4381 caaaaatatt cgtggtggaa atgaccaggc cataatgatc attcccgagc tggcacgggc
     4441 tacgggaatg acggacgcta tgcgcgctga ctttaggacg ttgagggcca tgagtgaaca
     4501 taccaggctg aatccagatc gtcgcatcga acgccttcgc atgttcaaca agcgcttaaa
     4561 gtcgtgtaaa cagagtgtag agacccttaa gtcctggaac atcgagctgg actccgcctt
     4621 ggtggagatt ccagcacgcg tgttgccacc ggaaaaaata ttattcggca accaaaaaat
     4681 attcgtatgc gacgctcgcg ccgattggac caatgagttt cgcacctgtt cgatgtttaa
     4741 aaacgtgcat atcaataggt ggtatgtgat cacaccgagt cgaaatctgc gcgagaccca
     4801 ggaatttgtg cagatgtgca tccgcacagc cagcagcatg aagatgaaca tctgcaatcc
     4861 aatatatgag gaaattccgg atgaccgcaa cggcacttac tcccaagcga tcgacaacgc
     4921 cgcagctaac gatccacaga tcgtgatggt tgtcatgaga tctccaaatg aagagaaata
     4981 tagctgcatt aaaaaacgca cgtgcgtgga cagaccggtg ccatcgcagg tggtgacact
     5041 aaaagttatc gcgcctcgac agcaaaaacc aactgggctg atgtcgatcg ccacaaaggt
     5101 ggttattcaa atgaacgcca aattgatggg agctccctgg caggtagtga tccccctcca
     5161 cggtctgatg actgttggtt tcgatgtctg ccattcaccg aagaataaga acaagtcata
     5221 tggggctttc gttgcaacca tggaccagaa ggagtctttc cgctacttct ccaccgtaaa
     5281 cgaacacatt aagggccaag agttgtccga gcagatgtcg gtgaacatgg cgtgcgccct
     5341 gaggtcgtat caggagcaac accgttcttt gccagagcgc attctcttct ttcgcgacgg
     5401 tgttggcgat ggtcagctct accaggtggt aaacagcgag gtgaacaccc taaaggacag
     5461 gctcgacgaa atttacaaat cagctggcaa acaggagggc tgtcgcatga catttataat
     5521 tgtatccaag cgcattaatt ctcgctactt tactgggcat cgcaacccag ttccgggcac
     5581 tgtagtcgat gacgttatta ccttgccaga gcgctacgac ttcttcctag tgtcccaggc
     5641 tgttcggata ggaactgtgt cgcctaccag ctacaatgtt atttctgaca acatgggact
     5701 aaacgccgat aagctgcaga tgctctccta taagatgacc catatgtact acaattacag
     5761 cggaaccata cgagtccccg ctgtctgcca ttacgcccac aaattggctt tcctcgtagc
     5821 cgaatccatt aatcgtgcgc cttcagcagg actgcagaat caattgtact ttttgtaaat
     5881 ccagcacagt ggcggccgct cgagtctaga gggcccgcgg ttcgaaggta agcctatccc
     5941 taaccctctc ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt
     6001 ttaaacccgc tgatcagcct cgactgtgcc ttctaaggcc tgagctcgct gatcagcctc
     6061 gatcgaggat ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     6121 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     6181 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     6241 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga
     6301 ttatgatcag tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg
     6361 cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag
     6421 gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga
     6481 cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg
     6541 tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg
     6601 ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg gccttcccca
     6661 ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca
     6721 ggcaggtaga tgacgaccat cagggacagc ttcaaggatc gctcgcggct cttaccagcc
     6781 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6841 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6901 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6961 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat
     7021 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     7081 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     7141 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     7201 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     7261 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     7321 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     7381 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7441 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7501 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7561 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7621 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7681 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7741 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7801 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7861 cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct
     7921 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7981 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     8041 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     8101 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     8161 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac
     8221 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     8281 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     8341 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     8401 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8461 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8521 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     8581 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     8641 gtcttcaagg aattaattct atattctaaa aacacaaatg atacttctaa
//
