LOCUS       sequence-407282-        9419 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cup expression for Schneider cell imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9419)
  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 9419)
  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: F23-pAc5.1 EGFP Cup
FEATURES             Location/Qualifiers
     source          1..9419
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..2517
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(2537..2539,2540..2542,2543..3253)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2537..2539,2540..2542,2543..3253)
                     /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(2843..2862)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3190..3211
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6677..6718
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6728..6745
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6768..6785)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6818..6952
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6842..6861)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6896..6915
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7553..8141)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7633..7652)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8312..9103,9104..9172))
                     /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     8935..8954
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9173..9277)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9345..9363
                     /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 accatcatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     2581 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     2641 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     2701 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     2761 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     2821 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     2881 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     2941 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     3001 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     3061 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     3121 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     3181 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     3241 cgagctgtac aagccaagct tgaattctgc agatatgcaa atggccgaag ctgagcaaga
     3301 aaatggcgct ggcgccctga aaattgccac caatgcgggt gcaaccgatc gtccagcgca
     3361 tcagcagctt ccgctgccag tcgaagatca gcaggatgag gtgctgaccc ccgctgagaa
     3421 gggcaagttc gagtaccctc cccctccacc gccaccaact ccagtccagg ctccgctagc
     3481 caccaaggcc accgctctga atgccagtca agagcatgac gacgacgagg ccaactccga
     3541 gaagtgggag gatccgtgcg ctccgccccc gcctcctccg ctgccgacta gcgctttcct
     3601 ggccaccgga ctgggctacc tcaagctgcc cgccttcaag ctgaaggacg cactggagaa
     3661 ggccatcacc aagttggagg ccaacaagcg gacgctaaag gccagccctg agtcttctcg
     3721 ttccatcaaa aacaagaatg tggtggccct ggagatgtta cccaggcgct caaacccaga
     3781 aacgatcggc gatggatcca tgttggcatc cacatccacg gctgttatgc tgcagaccaa
     3841 aaagcccgcc gttattgtgg agatggagag gcgttgtaag atcatcaatt tactggccaa
     3901 acagaatcaa atcctcgaat ccatttcggg ggaggccatt ccgatgcatg gaccctcgaa
     3961 gcatctgcac gaggacgagg gtctgaccct gcaagttctc tcggcccgag catccacacc
     4021 gtacacacag ccctccagca tgttgtcctg cacggccgtt agttgtgacc tggagcacga
     4081 ttcgccccgg aagcaagtgg ccagcaagga ggccgtgccg gagcagcagt ctagtcaggt
     4141 gcagcaaaag cgtccaccca gcacgggcat tcacaagcca ggaagtctgc gagcacccaa
     4201 ggctgttcgt cctactactg ctccggtggt cagcagcaag ccggtcaaga gctatacccg
     4261 ctcccgcctg atggacattc gcaacgggat gttcaatgcc ctgatgcaca ggtccaagga
     4321 gagctttgtg atgcctcgca tcgccacctg cgacgacatt gagttggagg gaaggcttcg
     4381 ccgcatgaat atttggcgca cctcggacgg cacaagattc cggacccgct ccaccacagc
     4441 taacttaaac atgaacaaca acaacaacaa tgagtgcatg cctgccttct tcaaaaacaa
     4501 aaataagccg aacctaatca gtgacgagtc catcatccag agccaaccgc cccagccgca
     4561 aacggagttc caagatccgg ccattgttaa ccaaagacgc attggcagcg gtcgcttaaa
     4621 ccattccaag tggggttaca acgacgaaga ctaccactcg taccacaatg gcaagtcgca
     4681 gcatatggag gaggtcaact cgaagaacag caaaaacatg acggtgctgc agttcttcga
     4741 caacggcgag atcagcagtc agcctcaaag acggccgaac actcctgtga tggggatgtc
     4801 tatcaatcga tccgagaacg acaccctgca ctcgaacgag tccagtgaag atttgagtcg
     4861 cgccaacgag aactatgtaa agcgcgtaat gtctggattc ctggttgtat cgaagcccaa
     4921 gtcacgagat gtcgaggata gacatcaccg gcggtacagg aaccagaacg aggagcctga
     4981 gtggtttagc tgtggaccca cgtctagact ggacactata gagttgtgtg ggttcgatga
     5041 ggacgaggag aaaatgctaa aggagggcaa caaaaaccac ggattaggcg aaacagagcg
     5101 agaaacgtca aaacaaaaga tggaccataa gtacaaatgg acgcatgcgg agccgatggg
     5161 tcgaaacaaa tacatgccca aacacgacac taacaataat cataacgtgg agaacatgaa
     5221 caatgttatg gctaccgagc atcagcaaca aaaggaggag aaacgtccag gcagtggccg
     5281 ttcgtttcag tttgataagt tcaatcaaag ccaacagaat tacgagagta gcagttatgt
     5341 taatcaccaa cagccgccgc aaacgcagcc tcagcaaatg cagcagcaat cgaacacaaa
     5401 cacgaacaac tctaaattca tgtcattctt tgccaatgag gggaactcct cgtcttcttc
     5461 gctgaacgag tttttcaagc aggccattaa ccagggtcat ggcaacaatc cggagcagcc
     5521 caaatcgttg ggccacatcg gacagatgcc ttcggtggat cagctggagg ccaaatggcg
     5581 tcgaaactca ctgaacaatg ttggcgagac tgccaacaag cagactgata acttccagaa
     5641 gctaattggg tctctttctt cggctaagcc acagtcacag gcggttggct atgatgccat
     5701 ctccaacttt ataatgcagc aacagcagta ccagcagcag caacagaagc agcacctcat
     5761 catacagcag cagcagcaac agaccgcctt tttggcgagc ctgcagctga aggcgatcct
     5821 cggccgggcc gacacccaat tgctactgct ccgtctgact aaaggggaaa tatccaagca
     5881 cggattgctg gtgcagttgg ccaacccacg cttgacggac atggatcgcg agaccatcac
     5941 tgctgtattg cagttcacca acacccagca gcagcagcag cagcacaagc agcagctgga
     6001 catgctctca agcacggtca tcgccagcca gttgcagaat ctccacaact tggccattgt
     6061 ccagcagact cttgcagcca ggcagcagcc gcagcataat ccacagacgc aggcgccgca
     6121 ccaactgtcg caggaggact tgcaagcgca tgctaatgtg ataatgcgaa acgccgtaat
     6181 gaagcgaaaa atggaggagc agacctcgaa gcttatcaat ggcggtgcca agcatcaggc
     6241 acagcagcaa tatctaaacc gtggccagca acgccaggct cgaccagatg ccaactcgaa
     6301 tgccttgctc catgccttga tctctggcgg tggcaataac cacgcctctg gttatcctat
     6361 gaatggacag ccgcagcagc atcattccaa tttgagattc ggtgataacc aaaacttcca
     6421 atcttttgag tccaaccagc cgcacttcgc cacgcagtac aagcagcaat accaacagtc
     6481 tcagcagcaa catccccatc agcagcccca acagctgaat tcgcttcatc agaacaacgc
     6541 tggagctgtc aatagcttca ataaggccca aatgcaagct caatcagcta tttccatgct
     6601 gcccaacagc ggggatgagt ttcattaaat ccagcacagt ggcggccgct cgagtctaga
     6661 gggcccgcgg ttcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgcg
     6721 taccggtcat catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc
     6781 ttctaaggcc tgagctcgct gatcagcctc gatcgaggat ccagacatga taagatacat
     6841 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     6901 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     6961 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     7021 gtaaaacctc tacaaatgtg gtatggctga ttatgatcag tcgaccgatg cccttgagag
     7081 ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta
     7141 tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt
     7201 tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg
     7261 gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg
     7321 agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt
     7381 tcgcgacgcg aggctggatg gccttcccca ttatgattct tctcgcttcc ggcggcatcg
     7441 ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat cagggacagc
     7501 ttcaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     7561 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     7621 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     7681 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     7741 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     7801 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     7861 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     7921 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     7981 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     8041 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     8101 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     8161 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     8221 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     8281 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     8341 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     8401 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     8461 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     8521 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     8581 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg
     8641 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     8701 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     8761 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     8821 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     8881 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata
     8941 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     9001 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     9061 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     9121 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     9181 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     9241 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     9301 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     9361 ggccctttcg tcttcaagaa ttaattctat attctaaaaa cacaaatgat acttctaaa
//
