LOCUS       sequence-407123-        8597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NOT3 expression in Schneider cells for imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8597)
  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 8597)
  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-mEGFP-NOT3 isoform A (HK55)
FEATURES             Location/Qualifiers
     source          1..8597
                     /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 with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2582..2603)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     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             5855..5896
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5906..5923
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5946..5963)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5996..6130
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6020..6039)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6074..6093
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6731..7319)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6811..6830)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7490..8281,8282..8350))
                     /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     8113..8132
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8351..8455)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8523..8541
                     /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 taattttctt 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 gtccaagctg agcaaagacc ccaacgagaa
     3181 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     3241 cgagctgtac aagccaagct tgaattctgc agatatggct gcgacgagaa aattgcaagg
     3301 cgaaattgat cgatgtttaa aaaaggtagc agagggcgtt gaaacgtttg aggatatttg
     3361 gaaaaaagta cacaatgcga caaacaccaa tcagaagcaa aaacatctgc aggagaagta
     3421 tgaggcagac ctcaaaaagg aaatcaagaa attgcaacga cttcgcgacc aaattaaatc
     3481 atggattgca tcagctgaaa ttaaggataa gagctccctt cttgagaatc gacgtctcat
     3541 tgaaacacaa atggaacgct ttaaggtggt tgagcgcgag acgaaaacaa aagcctactc
     3601 caaggaaggc ttgggggctg cacaaaagat ggatcctgcc cagcgtatta aagacgacgc
     3661 acgcaattgg ttaaccagct caataagttc ccttcaaata caaattgacc agtatgagag
     3721 cgagatcgaa tcgttgttgg caggtaaaaa gaaacgttta gatcgagaca agcaggagcg
     3781 aatggatgac ctacgaggaa agttggaccg ccataagttc cacataacaa agctggaaac
     3841 tctgcttagg ctgcttgaca atgacggcgt cgaagcagag caggtaaaca agatcaaaga
     3901 cgatgtcgaa tactacattg attcatcgca agagccagac tttgaggaaa atgagttcat
     3961 atacgatgat ataattggtc ttgatgaagt ggagctaagt ggcacggcta ccacggacag
     4021 caacaacagc aacgagacaa gcggttctcc aagcagtgtc acatccggag gcagtccatc
     4081 acagtcccct gtcacggttc aacaaatttt aaacacttcg tcgcaggggg cagctagtag
     4141 tggtagcagc gcagctagcg cagcactgtt tcagcagcag ctgacagcgg cacaatcgaa
     4201 tggcaacaat gtgggctacg cgagcgacac aagtgcagct tcttcgtctg ctacaacatc
     4261 tacagatcct gcaggcggca cagttgctgt aaattgtgtt ggagggctag ccgataaacg
     4321 caacaagagc agcgaaagca atgccctaaa attgaaacca caacctcatc aacttataaa
     4381 acctacccct gtacgagcca cggccaagct gcctctgagc agtgataccc aagtcaataa
     4441 gattgtcagt tcaactccca gcaaaaatca gcaacagctg cctacagcag cttcaatcgt
     4501 tgcaacctcc gctatgcaat cgcaaagcag tatcggaagt tgcagctcca ctggaggaac
     4561 gggagcatct caatcagctt ctagtggtaa caatcctggt aacaatcctg cggttcagcc
     4621 aaatgctcca acgcctggtc aaagcggcat tgccgctgca gcagcttcga ctaatgttgt
     4681 ttcggcgacg attgtgagca gcgccaatgt tcaaggccaa tctgtgatcc aaccaacacc
     4741 aacaattgca tttgctgcag tagcaaaaca caatacatca ctcctggaaa atggtcctgt
     4801 tttgcaacag caactagcag taactcctac tgtggcagct atcgtaggag ctggaacgca
     4861 ggcgcagcaa aaacatgtgc caccattatc caatcttcaa acaaattccc ctcacataca
     4921 aaatggtctt cctgtctcgg acagtactaa tgacaacagc tgcaatgtcg tcgatacgat
     4981 ttctctaaaa acaatggcac aggatgctat taatcgatct gcgatcgatc ctaattcact
     5041 gaatcagcag caaacaagca gcatcgatct gagacagccg cagtcgcaga aatcacttct
     5101 ccaacatttc aactctgaaa ccaacacgaa tcaacagcaa ttgacatcgc agcagcagca
     5161 acagcttcag aataactccc ttgctgcgac cacaggttct aacaacggaa cttcaactgg
     5221 tagtggtctc atgaatgtag caaacgctac tggtcagcct atcagcggca atgccaaaac
     5281 gcatgcgtgc caaccccaac agatggccac gacggaagcg cacattccca cactgcttgg
     5341 tgtgacacca cttgggccaa caccacttca aaaggagcac caaatgcagt tccaaatgat
     5401 ggaagctgcc tactatcacc tgccacagcc tatggacacg gaaaagctac aaacgtactt
     5461 tcatcgcgca ccagtgctca cgccatcgca ctatccccag gcacagatgc caatatatga
     5521 tacagttgag ttttatcaac gactctcaac ggagaccctt ttttttgtgt tctactatat
     5581 ggagggctcc aaggcccagt atctagcggc caaggcctta aaaaagcaga gctggcgttt
     5641 ccacacaaag tatatgatgt ggtttcaaag gcacgaggag ccgaagatta tcaacgatga
     5701 ttacgaacag ggtacgtaca tctattttga ttatgaaaag tggagtcagc gaaagaagga
     5761 gggattcacc tttgaataca agtacttaga ggacaaggag ctgaatatcc agcacagtgg
     5821 cggccgctcg agtctagagg gcccgcggtt cgaaggtaag cctatcccta accctctcct
     5881 cggtctcgat tctacgcgta ccggtcatca tcaccatcac cattgagttt aaacccgctg
     5941 atcagcctcg actgtgcctt ctaaggcctg agctcgctga tcagcctcga tcgaggatcc
     6001 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     6061 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     6121 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     6181 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcagtc
     6241 gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga
     6301 ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg
     6361 cagcgctctg ggtcattttc ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc
     6421 tgtcgcttgc ggtattcgga atcttgcacg ccctcgctca agccttcgtc actggtcccg
     6481 ccaccaaacg tttcggcgag aagcaggcca ttatcgccgg catggcggcc gacgcgctgg
     6541 gctacgtctt gctggcgttc gcgacgcgag gctggatggc cttccccatt atgattcttc
     6601 tcgcttccgg cggcatcggg atgcccgcgt tgcaggccat gctgtccagg caggtagatg
     6661 acgaccatca gggacagctt caaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6721 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6781 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6841 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6901 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6961 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     7021 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     7081 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7141 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     7201 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7261 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7321 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7381 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7441 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7501 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7561 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7621 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7681 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7741 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7801 ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7861 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7921 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7981 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     8041 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     8101 cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     8161 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     8221 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8281 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8341 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8401 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8461 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     8521 aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt aattctatat tctaaaaaca
     8581 caaatgatac ttctaaa
//
