LOCUS       sequence-425991-        8307 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human DDX21. C-terminal NanoLuc 
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8307)
  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 8307)
  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_DDX21:1-783
FEATURES             Location/Qualifiers
     source          1..8307
                     /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             3482..3994
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(4070..4088)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4125..4246
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4136..4155)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4190..4209
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4491..4848
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4491..4682
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4514..4534)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4699..4834
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4879..5916
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5895..5903
                     /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    5940..5988
                     /note="synthetic polyadenylation signal"
     promoter        6144..6248
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6249..6317,6318..7109)
                     /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(6467..6486)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7267..7855)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7347..7366)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7971..8254
                     /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 caccatgccg ggaaaactcc gtagtgacgc tggtttggaa tcagacaccg caatgaaaaa
     1141 aggggagaca ctgcgaaagc aaaccgagga gaaagagaaa aaagagaagc caaaatctga
     1201 taagactgaa gagatagcag aagaggaaga aactgttttc cccaaagcta aacaagttaa
     1261 aaagaaagca gagccttctg aagttgacat gaattctcct aaatccaaaa aggcaaaaaa
     1321 gaaagaggag ccatctcaaa atgacatttc tcctaaaacc aaaagtttga gaaagaaaaa
     1381 ggagcccatt gaaaagaaag tggtttcttc taaaaccaaa aaagtgacaa aaaatgagga
     1441 gccttctgag gaagaaatag atgctcctaa gcccaagaag atgaagaaag aaaaggaaat
     1501 gaatggagaa actagagaga aaagccccaa actgaagaat ggatttcctc atcctgaacc
     1561 ggactgtaac cccagtgaag ctgccagtga agaaagtaac agtgagatag agcaggaaat
     1621 acctgtggaa caaaaagaag gcgctttctc taattttccc atatctgaag aaactattaa
     1681 acttctcaaa ggccgaggag tgaccttcct atttcctata caagcaaaga cattccatca
     1741 tgtttacagc gggaaggact taattgcaca ggcacggaca ggaactggga agacattctc
     1801 ctttgccatc cctttgattg agaaacttca tggggaactg caagacagga agagaggccg
     1861 tgcccctcag gtactggttc ttgcacctac aagagagttg gcaaatcaag taagcaaaga
     1921 cttcagtgac atcacaaaaa agctgtcagt ggcttgtttt tatggtggaa ctccctatgg
     1981 aggtcaattt gaacgcatga ggaatgggat tgatatcctg gttggaacac caggtcgtat
     2041 caaagaccac atacagaatg gcaaactaga tctcaccaaa cttaagcatg ttgtcctgga
     2101 tgaagtggac cagatgttgg atatgggatt tgctgatcaa gtggaagaga ttttaagtgt
     2161 ggcatacaag aaagattctg aagacaatcc ccaaacattg cttttttctg caacttgccc
     2221 tcattgggta tttaatgttg ccaagaaata catgaaatct acatatgaac aggtggacct
     2281 gattggtaaa aagactcaga aaacggcaat aactgtggag catctggcta ttaagtgcca
     2341 ctggactcag agggcagcag ttattgggga tgtcatccga gtatatagtg gtcatcaagg
     2401 acgcactatc atcttttgtg aaaccaagaa agaagcccag gagctgtccc agaattcagc
     2461 tataaagcag gatgctcagt ccttgcatgg agacattcca cagaagcaaa gggaaatcac
     2521 cctgaaaggt tttagaaatg gtagttttgg agttttggtg gcaaccaatg ttgctgcacg
     2581 tgggttagac atccctgagg ttgatttggt tatacaaagc tctccaccaa aggatgtaga
     2641 gtcctacatt catcgatccg ggcggacagg cagagctgga aggacggggg tgtgcatctg
     2701 cttttatcag cacaaggaag aatatcagtt agtacaagtg gagcaaaaag cgggaattaa
     2761 gttcaaacga ataggtgttc cttctgcaac agaaataata aaagcttcca gcaaagatgc
     2821 catcaggctt ttggattccg tgcctcccac tgccattagt cacttcaaac aatcagctga
     2881 gaagctgata gaggagaagg gagctgtgga agctctggca gcagcactgg cccatatttc
     2941 aggtgccacg tccgtagacc agcgctcctt gatcaactca aatgtgggtt ttgtgaccat
     3001 gatcttgcag tgctcaattg aaatgccaaa tattagttat gcttggaaag aacttaaaga
     3061 gcagctgggc gaggagattg attccaaagt gaagggaatg gtttttctca aaggaaagct
     3121 gggtgtttgc tttgatgtac ctaccgcatc agtaacagaa atacaggaga aatggcatga
     3181 ttcacgacgc tggcagctct ctgtggccac agagcaacca gaactggaag gaccacggga
     3241 aggatatgga ggcttcaggg gacagcggga aggcagtcga ggcttcaggg gacagcggga
     3301 cggaaacaga agattcagag gacagcggga aggcagtaga ggcccgagag gacagcgatc
     3361 aggaggtggc aacaaaagta acagatccca aaacaaaggc cagaagcgga gtttcagtaa
     3421 agcatttggt caatctagat ttgggcccaa ttcctgcagg cgagctctcg gctcgagcgg
     3481 cgtcttcaca ctcgaagatt tcgttgggga ctggcgacag acagccggct acaacctgga
     3541 ccaagtcctt gaacagggag gtgtgtccag tttgtttcag aatctcgggg tgtccgtaac
     3601 tccgatccaa aggattgtcc tgagcggtga aaatgggctg aagatcgaca tccatgtcat
     3661 catcccgtat gaaggtctga gcggcgacca aatgggccag atcgaaaaaa tttttaaggt
     3721 ggtgtaccct gtggatgatc atcactttaa ggtgatcctg cactatggca cactggtaat
     3781 cgacggggtt acgccgaaca tgatcgacta tttcggacgg ccgtatgaag gcatcgccgt
     3841 gttcgacggc aaaaagatca ctgtaacagg gaccctgtgg aacggcaaca aaattatcga
     3901 cgagcgcctg atcaaccccg acggctccct gctgttccga gtaaccatca acggagtgac
     3961 cggctggcgg ctgtgcgaac gcattctggc gtaaggccgc gactctaggg catgcaagct
     4021 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     4081 taactagcat aaccccttgg ggcggccgct tcgagcagac atgataagat acattgatga
     4141 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     4201 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     4261 cattcatttt atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa
     4321 cctctacaaa tgtggtaaaa tcgaatttta acaaaatatt aacgcttaca atttcctgat
     4381 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgcg gatctgcgca
     4441 gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt ctgaggcgga
     4501 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     4561 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     4621 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4681 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4741 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4801 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcgattct
     4861 tctgacacta gcgccaccat gaagaagccc gaactcaccg ctaccagcgt tgaaaaattt
     4921 ctcatcgaga agttcgacag tgtgagcgac ctgatgcagt tgtcggaggg cgaagagagc
     4981 cgagccttca gcttcgatgt cggcggacgc ggctatgtac tgcgggtgaa tagctgcgct
     5041 gatggcttct acaaagaccg ctacgtgtac cgccacttcg ccagcgctgc actacccatc
     5101 cccgaagtgt tggacatcgg cgagttcagc gagagcctga catactgcat cagtagacgc
     5161 gcccaaggcg ttactctcca agacctcccc gaaacagagc tgcctgctgt gttacagcct
     5221 gtcgccgaag ctatggatgc tattgccgcc gccgacctca gtcaaaccag cggcttcggc
     5281 ccattcgggc cccaaggcat cggccagtac acaacctggc gggatttcat ttgcgccatt
     5341 gctgatcccc atgtctacca ctggcagacc gtgatggacg acaccgtgtc cgccagcgta
     5401 gctcaagccc tggacgaact gatgctgtgg gccgaagact gtcccgaggt gcgccacctc
     5461 gtccatgccg acttcggcag caacaacgtc ctgaccgaca acggccgcat caccgccgta
     5521 atcgactggt ccgaagctat gttcggggac agtcagtacg aggtggccaa catcttcttc
     5581 tggcggccct ggctggcttg catggagcag cagactcgct acttcgagcg ccggcatccc
     5641 gagctggccg gcagccctcg tctgcgagcc tacatgctgc gcatcggcct ggatcagctc
     5701 taccagagcc tcgtggacgg caacttcgac gatgctgcct gggctcaagg ccgctgcgat
     5761 gccatcgtcc gcagcggggc cggcaccgtc ggtcgcacac aaatcgctcg ccggagcgca
     5821 gccgtatgga ccgacggctg cgtcgaggtg ctggccgaca gcggcaaccg ccggcccagt
     5881 acacgaccgc gcgctaagga ggtaggtcga gtttagactc tagccatcac gatggccgca
     5941 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagcga
     6001 taaggatcct ctttgcgctt gcgttttccc ttgtccagat agcccagtag ctgacattca
     6061 tccggggtca gcaccgtttc tgcggactgg ctttctacgt aatggtttct tagacgtcag
     6121 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     6181 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     6241 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     6301 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     6361 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     6421 ttcgccccga agaacgtttt ccaatgatga gcactttcaa agttctgcta tgtggcgcgg
     6481 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     6541 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     6601 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     6661 caactatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6721 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     6781 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     6841 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     6901 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     6961 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     7021 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     7081 taggtgcctc actgattaag cattggtaat tcgaaatgac cgaccaagcg acgcccaacc
     7141 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     7201 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     7261 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     7321 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     7381 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7441 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7501 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7561 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7621 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7681 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     7741 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7801 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga
     7861 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7921 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttatagt ccggaaatac
     7981 aggaacgcac gctggatggc ccttcgctgg gatggtgaaa ccatgaaaaa tggcagcttc
     8041 agtggattaa gtgggggtaa tgtggcctgt accctctggt tgcataggta ttcatacggt
     8101 taaaatttat caggcgcgat tgcggcagtt tttcgggtgg tttgttgcca tttttacctg
     8161 tctgctgccg tgatcgcgct gaacgcgttt tagcggtgcg tacaattaag ggattatggt
     8221 aaatccactt actgtctgcc ctcgtagcca tcgagataaa ccgcagtact ccggccacga
     8281 tgcgtccggc gtagaggatc gagatct
//
