LOCUS       sequence-401731-        9020 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 9020)
  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 9020)
  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-NOT3 isoform A (HK100)
FEATURES             Location/Qualifiers
     source          1..9020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2568
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             2987..3694
                     /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(3132..3150)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3400..3419
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             6305..6346
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             6356..6373
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6396..6413)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6446..6580
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6470..6489)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6524..6543
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7202..7790)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7282..7301)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7961..8752,8753..8821))
                     /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     8584..8603
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8822..8926)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8994..9012
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cttcaaggaa ttaattctat attctaaaaa cacaaatgat acttctaaaa aaaatcatga
       61 atggcatcaa ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac
      121 atcatttttc cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg
      181 ccattaaagc tggctgatcg tcgccaaaca ccaaatacat atcaatatgt acattcgaga
      241 aagaagcgat caaagaagcg tcttcgggcg agtaggagaa tgcggaggag aaggagaacg
      301 agctgatcta gtatctctcc acaatccaat gccaactgac caactggcca tattcggagc
      361 aatttgaagc caatttccat cgcctggcga tcgctccatt cttggctata tgtttttcac
      421 cgttcccggg gccattttca aagactcgtc ggtaagataa gattgtgtca ctcgctgtct
      481 ctcttcattt gtcgaagaat gctgaggaat ttcgcgatga cgtcggcgag tattttgaag
      541 aatgagaata atttgtattt atacgaaaat cagttagtgg aattttctac aaaaacatgt
      601 tatctataga taattttgtt gcaaaatatg ttgactatga caaagattgt atgtatatac
      661 ctttaatgta ttctcatttt cttatgtatt tataatggca atgatgatac tgatgatatt
      721 ttaagatgat gccagaccac aggctgattt ctgcgtcttt tgccgaacgc agtgcatgtg
      781 cggttgttgt tttttggaat agtttcaatt ttcggactgt ccgctttgat ttcagtttct
      841 tggcttattc aaaaagcaaa gtaaagccaa aaaagcgaga tggcaatacc aaatgcggca
      901 aaacggtagt ggaaggaaag gggtgcgggg cagcggaagg aagggtgggg cggggcgtgg
      961 cggggtctgt ggctgggcgc gacgtcaccg acgttggagc cactcctttg accatgtgtg
     1021 cgtgtgtgta ttattcgtgt ctcgccactc gccggttgtt tttttctttt tatctcgctc
     1081 tctctagcgc catctcgtac gcatgctcaa cgcaccgcat gttgccgtgt cctttatgcg
     1141 tcattttggc tcgaaatagg caattattta aacaaagatt agtcaacgaa aacgctaaaa
     1201 taaataagtc tacaatatgg ttacttattg ccatgtgtgt gcagccaacg atagcaacaa
     1261 aagcaacaac acagtggctt tccctctttc actttttgtt tgcaagcgcg tgcgagcaag
     1321 acggcacgac cggcaaacgc aattacgctg acaaagagca gacgaagttt tggccgaaaa
     1381 acatcaaggc gcctgatacg aatgcatttg caataacaat tgcgatattt aatattgttt
     1441 atgaagctgt ttgacttcaa aacacacaaa aaaaaaaata aaacaaatta tttgaaagag
     1501 aattaggaat cggacagctt atcgttacgg gctaacagca caccgagacg aaatagctta
     1561 cctgacgtca cagcctctgg aagaactgcc gccaagcaga gagagagaga aaaagaggga
     1621 gagcagctta gaccgcatgt gcttgtgtgt gaggcgtctc tctcttcgtc tcctgtttgc
     1681 gcaaacgcat agactgcact gagaaaatcg attacctatt ttttatgaat gaatatttgc
     1741 actattacta ttcaaaacta ttaagatagc aatcacattc aatagccaaa tactatacca
     1801 cctgagcgat gcaacgaaat gatcaatttg agcaaaaatg ctgcatattt aggacggcat
     1861 cattatagaa atgcttcttg ctgtgtactt ttctctcgtc tggcagctgt ttcgccgtta
     1921 ttgttaaaac cggcttaagt taggtgtgtt ttctacgact agtgatgccc ctactagaag
     1981 atgtgtgttg cacaaatgtc cctgaataac caatttgaag tgcagatagc agtaaacgta
     2041 agctaatatg aatattattt aactgtaatg ttttaatatc gctggacatt actaataaac
     2101 ccactataaa cacatgtaca tatgtatgtt ttggcataca atgagtagtt ggggaaaaaa
     2161 tgtgtaaaag caccgtgacc atcacagcat aaagataacc agctgaagta tcgaatatga
     2221 gtaaccccca aattgaatca catgccgcaa ctgataggac ccatggaagt acactcttca
     2281 tggcgatata caagacacac acaagcacga acacccagtt gcggaggaaa ttctccgtaa
     2341 atgaaaaccc aatcggcgaa caattcatac ccatatatgg taaaagtttt gaacgcgact
     2401 tgagagcgga gagcattgcg gctgataagg ttttagcgct aagcgggctt tataaaacgg
     2461 gctgcgggac cagttttcat atcactaccg tttgagttct tgtgctgtgt ggatactcct
     2521 cccgacacaa agccgctcca tcagccagca gtcgtctaat ccagagaccc cggatcgggg
     2581 taccaacatg cacggtctcc aggatgatcc agatctgcag gctctgctga aaggttctca
     2641 gctcctcaaa gtcaaaagca gctcctggcg tcgtgaacgc ttctacaaac tgcaggagga
     2701 ctgcaaaacc atctggcagg aatcccgtaa agtaatgcgc tccccagagt ctcagctctt
     2761 cagcattgag gacattcagg aggtacgtat gggtcaccgt accgaaggtc tggagaaatt
     2821 cgctcgcgat atcccagaag atcgctgctt ctctatcgtc ttcaaagacc agcgcaacac
     2881 cctggatctg attgctccat ctccagcaga tgctcagcac tgggttcagg gtctccgcaa
     2941 aatcattcac cactccggtt ctatggacca gcgtcagaaa ggtaccatgg tgagcaaggg
     3001 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
     3061 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     3121 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat
     3181 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
     3241 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     3301 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
     3361 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
     3421 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
     3481 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac
     3541 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga
     3601 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg
     3661 ccactccacc ggcggcatgg acgagctgta caagccaagc ttgaattctg caacgtgcgc
     3721 tgcgacgaga aaattgcaag gcgaaattga tcgatgttta aaaaaggtag cagagggcgt
     3781 tgaaacgttt gaggatattt ggaaaaaagt acacaatgcg acaaacacca atcagaagca
     3841 aaaacatctg caggagaagt atgaggcaga cctcaaaaag gaaatcaaga aattgcaacg
     3901 acttcgcgac caaattaaat catggattgc atcagctgaa attaaggata agagctccct
     3961 tcttgagaat cgacgtctca ttgaaacaca aatggaacgc tttaaggtgg ttgagcgcga
     4021 gacgaaaaca aaagcctact ccaaggaagg cttgggggct gcacaaaaga tggatcctgc
     4081 ccagcgtatt aaagacgacg cacgcaattg gttaaccagc tcaataagtt cccttcaaat
     4141 acaaattgac cagtatgaga gcgagatcga atcgttgttg gcaggtaaaa agaaacgttt
     4201 agatcgagac aagcaggagc gaatggatga cctacgagga aagttggacc gccataagtt
     4261 ccacataaca aagctggaaa ctctgcttag gctgcttgac aatgacggcg tcgaagcaga
     4321 gcaggtaaac aagatcaaag acgatgtcga atactacatt gattcatcgc aagagccaga
     4381 ctttgaggaa aatgagttca tatacgatga tataattggt cttgatgaag tggagctaag
     4441 tggcacggct accacggaca gcaacaacag caacgagaca agcggttctc caagcagtgt
     4501 cacatccgga ggcagtccat cacagtcccc tgtcacggtt caacaaattt taaacacttc
     4561 gtcgcagggg gcagctagta gtggtagcag cgcagctagc gcagcactgt ttcagcagca
     4621 gctgacagcg gcacaatcga atggcaacaa tgtgggctac gcgagcgaca caagtgcagc
     4681 ttcttcgtct gctacaacat ctacagatcc tgcaggcggc acagttgctg taaattgtgt
     4741 tggagggcta gccgataaac gcaacaagag cagcgaaagc aatgccctaa aattgaaacc
     4801 acaacctcat caacttataa aacctacccc tgtacgagcc acggccaagc tgcctctgag
     4861 cagtgatacc caagtcaata agattgtcag ttcaactccc agcaaaaatc agcaacagct
     4921 gcctacagca gcttcaatcg ttgcaacctc cgctatgcaa tcgcaaagca gtatcggaag
     4981 ttgcagctcc actggaggaa cgggagcatc tcaatcagct tctagtggta acaatcctgg
     5041 taacaatcct gcggttcagc caaatgctcc aacgcctggt caaagcggca ttgccgctgc
     5101 agcagcttcg actaatgttg tttcggcgac gattgtgagc agcgccaatg ttcaaggcca
     5161 atctgtgatc caaccaacac caacaattgc atttgctgca gtagcaaaac acaatacatc
     5221 actcctggaa aatggtcctg ttttgcaaca gcaactagca gtaactccta ctgtggcagc
     5281 tatcgtagga gctggaacgc aggcgcagca aaaacatgtg ccaccattat ccaatcttca
     5341 aacaaattcc cctcacatac aaaatggtct tcctgtctcg gacagtacta atgacaacag
     5401 ctgcaatgtc gtcgatacga tttctctaaa aacaatggca caggatgcta ttaatcgatc
     5461 tgcgatcgat cctaattcac tgaatcagca gcaaacaagc agcatcgatc tgagacagcc
     5521 gcagtcgcag aaatcacttc tccaacattt caactctgaa accaacacga atcaacagca
     5581 attgacatcg cagcagcagc aacagcttca gaataactcc cttgctgcga ccacaggttc
     5641 taacaacgga acttcaactg gtagtggtct catgaatgta gcaaacgcta ctggtcagcc
     5701 tatcagcggc aatgccaaaa cgcatgcgtg ccaaccccaa cagatggcca cgacggaagc
     5761 gcacattccc acactgcttg gtgtgacacc acttgggcca acaccacttc aaaaggagca
     5821 ccaaatgcag ttccaaatga tggaagctgc ctactatcac ctgccacagc ctatggacac
     5881 ggaaaagcta caaacgtact ttcatcgcgc accagtgctc acgccatcgc actatcccca
     5941 ggcacagatg ccaatatatg atacagttga gttttatcaa cgactctcaa cggagaccct
     6001 tttttttgtg ttctactata tggagggctc caaggcccag tatctagcgg ccaaggcctt
     6061 aaaaaagcag agctggcgtt tccacacaaa gtatatgatg tggtttcaaa ggcacgagga
     6121 gccgaagatt atcaacgatg attacgaaca gggtacgtac atctattttg attatgaaaa
     6181 gtggagtcag cgaaagaagg agggattcac ctttgaatac aagtacttag aggacaagga
     6241 gctgaatgca tgagatatcc agcacagtgg cggccgctcg agtctagagg gcccgcggtt
     6301 cgaaggtaag cctatcccta accctctcct cggtctcgat tctacgcgta ccggtcatca
     6361 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctaaggcctg
     6421 agctcgctga tcagcctcga tcgaggatcc agacatgata agatacattg atgagtttgg
     6481 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     6541 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     6601 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     6661 caaatgtggt atggctgatt atgatcagtc gaccgatgcc cttgagagcc ttcaacccag
     6721 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
     6781 ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc
     6841 gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg
     6901 ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca
     6961 ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag
     7021 gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg atgcccgcgt
     7081 tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt caaggatcgc
     7141 tcgcggctct taccagccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     7201 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     7261 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     7321 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7381 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7441 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7501 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7561 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7621 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     7681 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7741 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7801 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7861 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7921 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7981 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     8041 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     8101 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     8161 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     8221 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag
     8281 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     8341 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8401 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8461 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8521 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac
     8581 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8641 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8701 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8761 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8821 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8881 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8941 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     9001 gtatcacgag gccctttcgt
//
