LOCUS       sequence-410787-        8999 bp    DNA     circular SYN 15-APR-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 8999)
  AUTHORS   Salgania HK, Metz J, Jeske M
  TITLE     ReLo is a simple and quick colocalization assay to identify and 
            characterize direct protein-protein interactions
  JOURNAL   bioRxiv 2022.03.04.482790
REFERENCE   2  (bases 1 to 8999)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2022.03.04.482790"
COMMENT     Sequence Label: pAc5.1-PH-mCherry-NOT3 isoform A (HK100)
FEATURES             Location/Qualifiers
     source          1..8999
                     /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             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(7181..7769)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7261..7280)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7940..8731,8732..8800))
                     /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     8563..8582
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8801..8905)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8973..8991
                     /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 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 caaggccagc
     7141 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     7201 ctgacgagca tcacaaaaat cgatgctcaa gtcagaggtg gcgaaacccg acaggactat
     7261 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     7321 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     7381 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     7441 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     7501 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     7561 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     7621 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     7681 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     7741 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7801 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7861 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7921 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7981 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     8041 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     8101 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     8161 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     8221 cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt
     8281 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     8341 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     8401 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     8461 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     8521 gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata
     8581 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8641 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8701 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8761 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8821 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8881 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     8941 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgt
//
