LOCUS       sequence-407176-        9833 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cup expression in Schneider cells for imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9833)
  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 9833)
  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-Cup isoform B (JM051)
FEATURES             Location/Qualifiers
     source          1..9833
                     /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(3245..3264)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3592..3613
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7091..7132
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             7142..7159
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7182..7199)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7232..7366
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7256..7275)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7310..7329
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7967..8555)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8047..8066)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8726..9517,9518..9586))
                     /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     9349..9368
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9587..9691)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9759..9777
                     /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 tgcaaatggc cgaagctgag caagaaaatg gcgctggcgc
     3721 cctgaaaatt gccaccaatg cgggtgcaac cgatcgtcca gcgcatcagc agcttccgct
     3781 gccagtcgaa gatcagcagg atgaggtgct gacccccgct gagaagggca agttcgagta
     3841 ccctccccct ccaccgccac caactccagt ccaggctccg ctagccacca aggccaccgc
     3901 tctgaatgcc agtcaagagc atgacgacga cgaggccaac tccgagaagt gggaggatcc
     3961 gtgcgctccg cccccgcctc ctccgctgcc gactagcgct ttcctggcca ccggactggg
     4021 ctacctcaag ctgcccgcct tcaagctgaa ggacgcactg gagaaggcca tcaccaagtt
     4081 ggaggccaac aagcggacgc taaaggccag ccctgagtct tctcgttcca tcaaaaacaa
     4141 gaatgtggtg gccctggaga tgttacccag gcgctcaaac ccagaaacga tcggcgatgg
     4201 atccatgttg gcatccacat ccacggctgt tatgctgcag accaaaaagc ccgccgttat
     4261 tgtggagatg gagaggcgtt gtaagatcat caatttactg gccaaacaga atcaaatcct
     4321 cgaatccatt tcgggggagg ccattccgat gcatggaccc tcgaagcatc tgcacgagga
     4381 cgagggtctg accctgcaag ttctctcggc ccgagcatcc acaccgtaca cacagccctc
     4441 cagcatgttg tcctgcacgg ccgttagttg tgacctggag cacgattcgc cccggaagca
     4501 agtggccagc aaggaggccg tgccggagca gcagtctagt caggtgcagc aaaagcgtcc
     4561 acccagcacg ggcattcaca agccaggaag tctgcgagca cccaaggctg ttcgtcctac
     4621 tactgctccg gtggtcagca gcaagccggt caagagctat acccgctccc gcctgatgga
     4681 cattcgcaac gggatgttca atgccctgat gcacaggtcc aaggagagct ttgtgatgcc
     4741 tcgcatcgcc acctgcgacg acattgagtt ggagggaagg cttcgccgca tgaatatttg
     4801 gcgcacctcg gacggcacaa gattccggac ccgctccacc acagctaact taaacatgaa
     4861 caacaacaac aacaatgagt gcatgcctgc cttcttcaaa aacaaaaata agccgaacct
     4921 aatcagtgac gagtccatca tccagagcca accgccccag ccgcaaacgg agttccaaga
     4981 tccggccatt gttaaccaaa gacgcattgg cagcggtcgc ttaaaccatt ccaagtgggg
     5041 ttacaacgac gaagactacc actcgtacca caatggcaag tcgcagcata tggaggaggt
     5101 caactcgaag aacagcaaaa acatgacggt gctgcagttc ttcgacaacg gcgagatcag
     5161 cagtcagcct caaagacggc cgaacactcc tgtgatgggg atgtctatca atcgatccga
     5221 gaacgacacc ctgcactcga acgagtccag tgaagatttg agtcgcgcca acgagaacta
     5281 tgtaaagcgc gtaatgtctg gattcctggt tgtatcgaag cccaagtcac gagatgtcga
     5341 ggatagacat caccggcggt acaggaacca gaacgaggag cctgagtggt ttagctgtgg
     5401 acccacgtct agactggaca ctatagagtt gtgtgggttc gatgaggacg aggagaaaat
     5461 gctaaaggag ggcaacaaaa accacggatt aggcgaaaca gagcgagaaa cgtcaaaaca
     5521 aaagatggac cataagtaca aatggacgca tgcggagccg atgggtcgaa acaaatacat
     5581 gcccaaacac gacactaaca ataatcataa cgtggagaac atgaacaatg ttatggctac
     5641 cgagcatcag caacaaaagg aggagaaacg tccaggcagt ggccgttcgt ttcagtttga
     5701 taagttcaat caaagccaac agaattacga gagtagcagt tatgttaatc accaacagcc
     5761 gccgcaaacg cagcctcagc aaatgcagca gcaatcgaac acaaacacga acaactctaa
     5821 attcatgtca ttctttgcca atgaggggaa ctcctcgtct tcttcgctga acgagttttt
     5881 caagcaggcc attaaccagg gtcatggcaa caatccggag cagcccaaat cgttgggcca
     5941 catcggacag atgccttcgg tggatcagct ggaggccaaa tggcgtcgaa actcactgaa
     6001 caatgttggc gagactgcca acaagcagac tgataacttc cagaagctaa ttgggtctct
     6061 ttcttcggct aagccacagt cacaggcggt tggctatgat gccatctcca actttataat
     6121 gcagcaacag cagtaccagc agcagcaaca gaagcagcac ctcatcatac agcagcagca
     6181 gcaacagacc gcctttttgg cgagcctgca gctgaaggcg atcctcggcc gggccgacac
     6241 ccaattgcta ctgctccgtc tgactaaagg ggaaatatcc aagcacggat tgctggtgca
     6301 gttggccaac ccacgcttga cggacatgga tcgcgaggcc atcactgctg tattgcagtt
     6361 caccaacacc cagcagcagc agcagcagca caagcagcag ctggacatgc tctcaagcac
     6421 ggtcatcgcc agccagttgc agaatctcca caacttggcc attgtccagc agactcttgc
     6481 agccaggcag cagccgcagc ataatccaca gacgcaggcg ccgcaccaac tgtcgcagga
     6541 ggacttgcaa gcgcatgcta atgtgataat gcgaaacgcc gtaatgaagc gaaaaatgga
     6601 ggagcagacc tcgaagctta tcaatggcgg tgccaagcat caggcacagc agcaatatct
     6661 aaaccgtggc cagcaacgcc aggctcgacc agatgccaac tcgaatgcct tgctccatgc
     6721 cttgatctct ggcggtggca ataaccacgc ctctggttat cctatgaatg gacagccgca
     6781 gcagcatcat tccaatttga gattcggtga taaccaaaac ttccaatctt ttgagtccaa
     6841 ccagccgcac ttcgccacgc agtacaagca gcaataccaa cagtctcagc agcaacatcc
     6901 ccatcagcag ccccaacagc tgaattcgct tcatcagaac aacgctggag ctgtcaatag
     6961 cttcaataag gcccaaatgc aagctcaatc agctatttcc atgctgccca acagcgggga
     7021 tgagtttcat taagcatgag atatccagca cagtggcggc cgctcgagtc tagagggccc
     7081 gcggttcgaa ggtaagccta tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg
     7141 tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctaa
     7201 ggcctgagct cgctgatcag cctcgatcga ggatccagac atgataagat acattgatga
     7261 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     7321 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     7381 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     7441 cctctacaaa tgtggtatgg ctgattatga tcagtcgacc gatgcccttg agagccttca
     7501 acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg
     7561 tcttctttat catgcaactc gtaggacagg tgccggcagc gctctgggtc attttcggcg
     7621 aggaccgctt tcgctggagc gcgacgatga tcggcctgtc gcttgcggta ttcggaatct
     7681 tgcacgccct cgctcaagcc ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc
     7741 aggccattat cgccggcatg gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga
     7801 cgcgaggctg gatggccttc cccattatga ttcttctcgc ttccggcggc atcgggatgc
     7861 ccgcgttgca ggccatgctg tccaggcagg tagatgacga ccatcaggga cagcttcaag
     7921 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     7981 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     8041 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     8101 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     8161 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     8221 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     8281 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     8341 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     8401 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     8461 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     8521 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     8581 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     8641 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     8701 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     8761 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     8821 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     8881 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     8941 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     9001 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac
     9061 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     9121 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     9181 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     9241 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     9301 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc
     9361 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     9421 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     9481 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     9541 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     9601 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     9661 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     9721 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     9781 ttcgtcttca agaattaatt ctatattcta aaaacacaaa tgatacttct aaa
//
