LOCUS       sequence-407193-        8222 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NOT2 expression in Schneider cells for imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8222)
  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 8222)
  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-mCherry-NOT2 isoform A (HK99)
FEATURES             Location/Qualifiers
     source          1..8222
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2565
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             2984..3691
                     /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(3129..3147)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3397..3416
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             5528..5569
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5579..5596
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5619..5636)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5669..5803
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5693..5712)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5747..5766
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(6404..6992)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6484..6503)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7163..7954,7955..8023))
                     /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     7786..7805
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8024..8128)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8196..8214
                     /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 caacatgcac ggtctccagg atgatccaga tctgcaggct ctgctgaaag gttctcagct
     2641 cctcaaagtc aaaagcagct cctggcgtcg tgaacgcttc tacaaactgc aggaggactg
     2701 caaaaccatc tggcaggaat cccgtaaagt aatgcgctcc ccagagtctc agctcttcag
     2761 cattgaggac attcaggagg tacgtatggg tcaccgtacc gaaggtctgg agaaattcgc
     2821 tcgcgatatc ccagaagatc gctgcttctc tatcgtcttc aaagaccagc gcaacaccct
     2881 ggatctgatt gctccatctc cagcagatgc tcagcactgg gttcagggtc tccgcaaaat
     2941 cattcaccac tccggttcta tggaccagcg tcagaaaggt accatggtga gcaagggcga
     3001 ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc
     3061 cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     3121 ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct
     3181 gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga
     3241 ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     3301 cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa
     3361 ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat
     3421 gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat
     3481 caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta
     3541 caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat
     3601 cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca
     3661 ctccaccggc ggcatggacg agctgtacaa gccaagcttg aattctgcaa cgtgcatggc
     3721 gaatttaaat tttcaacaac ccccgagaag tatagcgaac gcggcgttag ctggccggac
     3781 aacaggcgga ttcggtggta gctccctggc tggccatgtg acgcccacgt cgggcatgtt
     3841 tcaaaccgac ttcgccaact cctatccagg aacggcgaac tacggacagg cgccgcagca
     3901 gcagcaacaa cagcagcagc agccccaact gtcgccgaac cggaatgccc agctgtcggt
     3961 cggcggaccc gccatctcga gcggcaaccg caacgccaac ctcttcggcc agcgtcagtt
     4021 tgtggagcgg cgagccatgc aaggattggg cagtggcccc atgtcgaaca tgggcaactt
     4081 catgcagacg ggtcgcggcg gctacggaac gggcggaggc ggtggcggtc ctttgaataa
     4141 cttccacgtg ttcggcggcg gcggtggtag cgacacctcc acgcctgccc tgctcgatcc
     4201 cacggagttc ccctcgctga cgaatgcgcg cggccagaac gaccagacac tgccccaatc
     4261 gaatccgctc cagccgccgg gcagcaaacc ctatggtaat ttctttacct cttttggcat
     4321 ggtgaagcaa ccgacgtcag agcaatcgga gttcacgatg tctaacgagg acttccctgc
     4381 gttaccgggc acccaaaact cggacggcac aacgaacgcg gtggggagcg tcgccggaac
     4441 aggaggctcc ggaggtgcgt ccacagagaa ccacttggac ggcacggaaa agccgatgaa
     4501 ttcgatagtg gtcagcggat cagcgagcgg cagcagcggt tccaacgtcg gcgttgtagg
     4561 cggcaatggt ctcggtgccg ttggcagtgg tatagggggc ttggcagtag gcggcggcgg
     4621 aggagctggt agctcaggag gcggaggagt cggcggcaat gcagccagcg gcgttgtcgg
     4681 cgggtcccat gtaggactgg tgggcagtaa cagcggcatt ggcggcgtca acagcgtatc
     4741 caacagcaat gccatgatgg gcgtaggcgg cggcctgggg agtggaagcg gctcgtctgg
     4801 gagcggagcg ggcggcgagc atctgaacga taattccagc aacgataaac ttgtaaagag
     4861 cggcgtgcaa acttcgccag atgggaaggt cacaaacata ccagcgacca tggtgaacaa
     4921 ccagttcggc atggttggcc tgctgacgtt cattcgggct gcagaaacgg atcccaatct
     4981 ggtgacgctc tccttgggta cggatctcac gggtttaggc ctgaacctga actcgcagga
     5041 gagcttgcat acaaccttcg ctggcccctt cgtagcacag ccctgcagag cacaagacgt
     5101 tgagttcaac gtgccacccg aatatctgat taactttgcg ataagagaca aactaacagc
     5161 gccggtactg aaaaagctgc aagaagacct gctctttttc cttttttaca cgaacatcgg
     5221 cgatatgatg caattgatgg cagctgctga attgcatagt cgcgagtggc gataccacgt
     5281 ggaggagaaa atatggataa ctcgaattcc tggcattgat caatatgaaa aaaatggtac
     5341 aaaagagcgc ggcaccttct actactttga tgcgcaaagt tggaaacgtt tgtccaaggt
     5401 tttccaaata gatgccgaga aattagataa atgtcccaat ataagtgcgt ttatgaatgg
     5461 acagtctgta gcatgagata tccagcacag tggcggccgc tcgagtctag agggcccgcg
     5521 gttcgaaggt aagcctatcc ctaaccctct cctcggtctc gattctacgc gtaccggtca
     5581 tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctaaggc
     5641 ctgagctcgc tgatcagcct cgatcgagga tccagacatg ataagataca ttgatgagtt
     5701 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
     5761 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
     5821 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
     5881 ctacaaatgt ggtatggctg attatgatca gtcgaccgat gcccttgaga gccttcaacc
     5941 cagtcagctc cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct
     6001 tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg
     6061 accgctttcg ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc
     6121 acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc gagaagcagg
     6181 ccattatcgc cggcatggcg gccgacgcgc tgggctacgt cttgctggcg ttcgcgacgc
     6241 gaggctggat ggccttcccc attatgattc ttctcgcttc cggcggcatc gggatgcccg
     6301 cgttgcaggc catgctgtcc aggcaggtag atgacgacca tcagggacag cttcaaggcc
     6361 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6421 cccctgacga gcatcacaaa aatcgatgct caagtcagag gtggcgaaac ccgacaggac
     6481 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6541 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6601 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6661 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6721 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6781 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6841 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6901 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6961 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     7021 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     7081 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     7141 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7201 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7261 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7321 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7381 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7441 cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct
     7501 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7561 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7621 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7681 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7741 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac
     7801 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7861 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7921 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7981 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     8041 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     8101 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     8161 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     8221 gt
//
