LOCUS       sequence-426002-        6774 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human EXOSC6. C-terminal NanoLuc
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6774)
  AUTHORS   Ackloo S, Li F, Szewczyk M, Seitova A, Loppnau P, Zeng H, Xu J, 
            Ahmad S, Arnautova YA, Baghaie AJ, Beldar S, Bolotokova A, Centrella
            PA, Chau I, Clark MA, Cuozzo JW, Dehghani-Tafti S, Disch JS, Dong A,
            Dumas A, Feng JA, Ghiabi P, Gibson E, Gilmer J, Goldman B, Green SR,
            Guie MA, Guilinger JP, Harms N, Herasymenko O, Houliston S, 
            Hutchinson A, Kearnes S, Keefe AD, Kimani SW, Kramer T, Kutera M, 
            Kwak HA, Lento C, Li Y, Liu J, Loup J, Machado RAC, Mulhern CJ, 
            Perveen S, Righetto GL, Riley P, Shrestha S, Sigel EA, Silva M, 
            Sintchak MD, Slakman BL, Taylor RD, Thompson J, Torng W, 
            Underkoffler C, von Rechenberg M, Walsh RT, Watson I, Wilson DJ, 
            Wolf E, Yadav M, Yazdi AK, Zhang J, Zhang Y, Santhakumar V, Edwards 
            AM, Barsyte-Lovejoy D, Schapira M, Brown PJ, Halabelian L, 
            Arrowsmith CH
  TITLE     A Target Class Ligandability Evaluation of WD40 Repeat-Containing 
            Proteins.
  JOURNAL   J Med Chem. 2025 Jan 23;68(2):1092-1112. doi: 
            10.1021/acs.jmedchem.4c02010. Epub 2024 Nov 4.
  PUBMED    39495097
REFERENCE   2  (bases 1 to 6774)
  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"; doi: 
            "10.1021/acs.jmedchem.4c02010"; journalName: "J Med Chem"; date: 
            "2025-01-23- 23"; volume: "68"; issue: "2"; pages: "1092-1112"
COMMENT     Sequence Label: pNLF1-C_EXOSC6:1-272
FEATURES             Location/Qualifiers
     source          1..6774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1949..2461
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2537..2555)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2592..2713
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2603..2622)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2657..2676
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2958..3315
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2958..3149
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2981..3001)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3166..3301
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3346..4383
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4362..4370
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    4407..4455
                     /note="synthetic polyadenylation signal"
     promoter        4611..4715
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4716..4784,4785..5576)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4934..4953)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5734..6322)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5814..5833)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6438..6721
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcaaa gcgatcgctt ccgaattcgc
     1081 caccatgcct ggggatcacc gccgcatccg cggccctgaa gaatcgcagc cgccgcagct
     1141 gtacgcggcc gacgaggagg aggcgcccgg cacccgcgac ccaacgcggc tacggcccgt
     1201 gtacgcgcgc gccgggctgc tgagccaggc caagggctcg gcctacctgg aggcgggagg
     1261 caccaaggtg ctgtgtgccg tgtcgggccc gcgacaggcc gagggcggcg agcgcggcgg
     1321 cggcccggcc ggagcaggcg gcgaggcccc ggccgcgctg cgcggtcgcc tgctctgcga
     1381 cttccgccgc gcacccttcg cgggccgccg gcgccgcgct cccccgggcg gctgcgagga
     1441 gcgtgagctg gcgctggcgc tgcaggaggc gctggagccg gctgtgcgcc tgggccgcta
     1501 cccgcgcgcg cagctcgagg tgtcggcgct gctgctggag gacggtggct cggccctggc
     1561 cgccgcgctc accgccgccg cgctcgccct ggccgacgcg ggcgtggaga tgtacgacct
     1621 ggtggtgggc tgcggcctca gcctcgcgcc ggggcccgcg cccacctggc tcctggaccc
     1681 cacgcggctc gaggaagagc gcgccgccgc cggcctcacc gtggcgctca tgcctgtgct
     1741 gaatcaggtg gccgggctgc tgggcagcgg cgagggcggc ctgacagaga gctgggcgga
     1801 ggccgtacgc ctgggcctcg agggctgcca gcgcctctac cccgtgctgc agcagagcct
     1861 ggtgcgggcc gcccgccgca ggggcgccgc cgcccagccc tctagatttg ggcccaattc
     1921 ctgcaggcga gctctcggct cgagcggcgt cttcacactc gaagatttcg ttggggactg
     1981 gcgacagaca gccggctaca acctggacca agtccttgaa cagggaggtg tgtccagttt
     2041 gtttcagaat ctcggggtgt ccgtaactcc gatccaaagg attgtcctga gcggtgaaaa
     2101 tgggctgaag atcgacatcc atgtcatcat cccgtatgaa ggtctgagcg gcgaccaaat
     2161 gggccagatc gaaaaaattt ttaaggtggt gtaccctgtg gatgatcatc actttaaggt
     2221 gatcctgcac tatggcacac tggtaatcga cggggttacg ccgaacatga tcgactattt
     2281 cggacggccg tatgaaggca tcgccgtgtt cgacggcaaa aagatcactg taacagggac
     2341 cctgtggaac ggcaacaaaa ttatcgacga gcgcctgatc aaccccgacg gctccctgct
     2401 gttccgagta accatcaacg gagtgaccgg ctggcggctg tgcgaacgca ttctggcgta
     2461 aggccgcgac tctagggcat gcaagctgat ccggctgcta acaaagcccg aaaggaagct
     2521 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ggccgcttcg
     2581 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2641 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2701 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     2761 gtgggaggtt tttttaagca agtaaaacct ctacaaatgt ggtaaaatcg aattttaaca
     2821 aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     2881 ttcacaccgc atacgcggat ctgcgcagca ccatggcctg aaataacctc tgaaagagga
     2941 acttggttag gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     3001 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3061 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3121 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3181 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3241 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3301 ctaggctttt gcaaaaagct cgattcttct gacactagcg ccaccatgaa gaagcccgaa
     3361 ctcaccgcta ccagcgttga aaaatttctc atcgagaagt tcgacagtgt gagcgacctg
     3421 atgcagttgt cggagggcga agagagccga gccttcagct tcgatgtcgg cggacgcggc
     3481 tatgtactgc gggtgaatag ctgcgctgat ggcttctaca aagaccgcta cgtgtaccgc
     3541 cacttcgcca gcgctgcact acccatcccc gaagtgttgg acatcggcga gttcagcgag
     3601 agcctgacat actgcatcag tagacgcgcc caaggcgtta ctctccaaga cctccccgaa
     3661 acagagctgc ctgctgtgtt acagcctgtc gccgaagcta tggatgctat tgccgccgcc
     3721 gacctcagtc aaaccagcgg cttcggccca ttcgggcccc aaggcatcgg ccagtacaca
     3781 acctggcggg atttcatttg cgccattgct gatccccatg tctaccactg gcagaccgtg
     3841 atggacgaca ccgtgtccgc cagcgtagct caagccctgg acgaactgat gctgtgggcc
     3901 gaagactgtc ccgaggtgcg ccacctcgtc catgccgact tcggcagcaa caacgtcctg
     3961 accgacaacg gccgcatcac cgccgtaatc gactggtccg aagctatgtt cggggacagt
     4021 cagtacgagg tggccaacat cttcttctgg cggccctggc tggcttgcat ggagcagcag
     4081 actcgctact tcgagcgccg gcatcccgag ctggccggca gccctcgtct gcgagcctac
     4141 atgctgcgca tcggcctgga tcagctctac cagagcctcg tggacggcaa cttcgacgat
     4201 gctgcctggg ctcaaggccg ctgcgatgcc atcgtccgca gcggggccgg caccgtcggt
     4261 cgcacacaaa tcgctcgccg gagcgcagcc gtatggaccg acggctgcgt cgaggtgctg
     4321 gccgacagcg gcaaccgccg gcccagtaca cgaccgcgcg ctaaggaggt aggtcgagtt
     4381 tagactctag ccatcacgat ggccgcaata aaatatcttt attttcatta catctgtgtg
     4441 ttggtttttt gtgtgaatcg atagcgataa ggatcctctt tgcgcttgcg ttttcccttg
     4501 tccagatagc ccagtagctg acattcatcc ggggtcagca ccgtttctgc ggactggctt
     4561 tctacgtaat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4621 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4681 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4741 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4801 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     4861 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     4921 ctttcaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     4981 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     5041 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     5101 ataacactgc ggccaactta cttctgacaa ctatcggagg accgaaggag ctaaccgctt
     5161 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     5221 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     5281 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     5341 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     5401 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     5461 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     5521 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaattcg
     5581 aaatgaccga ccaagcgacg cccaaccggt atcagctcac tcaaaggcgg taatacggtt
     5641 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5701 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5761 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5821 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5881 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5941 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6001 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6061 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6121 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     6181 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6241 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6301 gcgcagaaaa aaaggatttc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6361 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6421 ctagatcctt ttatagtccg gaaatacagg aacgcacgct ggatggccct tcgctgggat
     6481 ggtgaaacca tgaaaaatgg cagcttcagt ggattaagtg ggggtaatgt ggcctgtacc
     6541 ctctggttgc ataggtattc atacggttaa aatttatcag gcgcgattgc ggcagttttt
     6601 cgggtggttt gttgccattt ttacctgtct gctgccgtga tcgcgctgaa cgcgttttag
     6661 cggtgcgtac aattaaggga ttatggtaaa tccacttact gtctgccctc gtagccatcg
     6721 agataaaccg cagtactccg gccacgatgc gtccggcgta gaggatcgag atct
//
