LOCUS       sequence-431698-        7056 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of full-length human WDR55. N-terminal NanoLuc 
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7056)
  AUTHORS   Ackloo S, Li F, Szewczyk M, Seitova A, Loppnau P, Zeng H, Xu J, 
            Ahmad S, Arnautova Y, Baghaie A, Beldar S, Bolotokova A, Centrella 
            P, Chau I, Clark M, Cuozzo J, Dehghani-Tafti S, Disch J, Dong A, 
            Dumas A, Feng J, Ghiabi P, Gibson E, Gilmer J, Goldman B, Green S, 
            Guie M, Guilinger J, Harms N, Herasymenko O, Houliston S, Hutchinson
            A, Kearnes S, Keefe A, Kimani S, Kramer T, Kutera M, Kwak H, Lento 
            C, Li Y, Liu J, Loup J, Machado R, Mulhern C, Perveen S, Righetto G,
            Riley P, Shrestha S, Sigel E, Silva M, Sintchak M, Slakman B, Taylor
            R, Thompson J, Torng W, Underkoffler C, Rechenberg M, Watson I, 
            Wilson D, Wolf E, Yadav M, Yazdi A, Zhang J, Zhang Y, Santhakumar V,
            Edwards A, Barsyte-Lovejoy D, Schapira M, Brown P, Halabelian L, 
            Arrowsmith C
  TITLE     A resource to enable chemical biology and drug discovery of WDR 
            Proteins
  JOURNAL   bioRxiv 2024.03.03.583197
REFERENCE   2  (bases 1 to 7056)
  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 
            2024.03.03.583197"
COMMENT     Sequence Label: pNLF1-N_WDR55:1-383
FEATURES             Location/Qualifiers
     source          1..7056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    332..615
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
     rep_origin      731..1319
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1220..1239
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1477..2268,2269..2337))
                     /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     2100..2119
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2338..2442)
                     /gene="bla"
                     /label=AmpR promoter
     polyA_signal    2598..2646
                     /note="synthetic polyadenylation signal"
     CDS             complement(2670..3707)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             2683..2691
                     /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)"
     promoter        complement(3738..4095)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3752..3887
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3904..4095
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4052..4072
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    4340..4461
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4377..4396)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4431..4450
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4493..4511
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(5758..6270)
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     complement(6283..6302)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6284..6302)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6344..6363)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          6346..6478
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(6614..6817)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6643..6663)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        join(6818..7056,1..141)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg
       61 taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat
      121 aactagtcaa taatcaatgc caacatggcg gtcatattgg acatgagcca atataaatgt
      181 acatattatg atatagatac aacgtatgca atggccaata gccaatattg atttatgcta
      241 tataaccaat gaataatatg gctaatggcc aatattgaag atctcgatcc tctacgccgg
      301 acgcatcgtg gccggagtac tgcggtttat ctcgatggct acgagggcag acagtaagtg
      361 gatttaccat aatcccttaa ttgtacgcac cgctaaaacg cgttcagcgc gatcacggca
      421 gcagacaggt aaaaatggca acaaaccacc cgaaaaactg ccgcaatcgc gcctgataaa
      481 ttttaaccgt atgaatacct atgcaaccag agggtacagg ccacattacc cccacttaat
      541 ccactgaagc tgccattttt catggtttca ccatcccagc gaagggccat ccagcgtgcg
      601 ttcctgtatt tccggactat aaaaggatct aggtgaagat cctttttgat aatctcatga
      661 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
      721 aaggatcttc ttgaaatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
      781 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      841 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
      901 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
      961 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     1021 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     1081 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     1141 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     1201 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     1261 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     1321 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     1381 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     1441 ataccggttg ggcgtcgctt ggtcggtcat ttcgaattac caatgcttaa tcagtgaggc
     1501 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     1561 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     1621 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     1681 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     1741 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     1801 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     1861 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     1921 agttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     1981 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     2041 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     2101 taataccgcg ccacatagca gaactttgaa agtgctcatc attggaaaac gttcttcggg
     2161 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     2221 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     2281 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     2341 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     2401 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     2461 gccacctgac gtctaagaaa ccattacgta gaaagccagt ccgcagaaac ggtgctgacc
     2521 ccggatgaat gtcagctact gggctatctg gacaagggaa aacgcaagcg caaagaggat
     2581 ccttatcgct atcgattcac acaaaaaacc aacacacaga tgtaatgaaa ataaagatat
     2641 tttattgcgg ccatcgtgat ggctagagtc taaactcgac ctacctcctt agcgcgcggt
     2701 cgtgtactgg gccggcggtt gccgctgtcg gccagcacct cgacgcagcc gtcggtccat
     2761 acggctgcgc tccggcgagc gatttgtgtg cgaccgacgg tgccggcccc gctgcggacg
     2821 atggcatcgc agcggccttg agcccaggca gcatcgtcga agttgccgtc cacgaggctc
     2881 tggtagagct gatccaggcc gatgcgcagc atgtaggctc gcagacgagg gctgccggcc
     2941 agctcgggat gccggcgctc gaagtagcga gtctgctgct ccatgcaagc cagccagggc
     3001 cgccagaaga agatgttggc cacctcgtac tgactgtccc cgaacatagc ttcggaccag
     3061 tcgattacgg cggtgatgcg gccgttgtcg gtcaggacgt tgttgctgcc gaagtcggca
     3121 tggacgaggt ggcgcacctc gggacagtct tcggcccaca gcatcagttc gtccagggct
     3181 tgagctacgc tggcggacac ggtgtcgtcc atcacggtct gccagtggta gacatgggga
     3241 tcagcaatgg cgcaaatgaa atcccgccag gttgtgtact ggccgatgcc ttggggcccg
     3301 aatgggccga agccgctggt ttgactgagg tcggcggcgg caatagcatc catagcttcg
     3361 gcgacaggct gtaacacagc aggcagctct gtttcgggga ggtcttggag agtaacgcct
     3421 tgggcgcgtc tactgatgca gtatgtcagg ctctcgctga actcgccgat gtccaacact
     3481 tcggggatgg gtagtgcagc gctggcgaag tggcggtaca cgtagcggtc tttgtagaag
     3541 ccatcagcgc agctattcac ccgcagtaca tagccgcgtc cgccgacatc gaagctgaag
     3601 gctcggctct cttcgccctc cgacaactgc atcaggtcgc tcacactgtc gaacttctcg
     3661 atgagaaatt tttcaacgct ggtagcggtg agttcgggct tcttcatggt ggcgctagtg
     3721 tcagaagaat cgagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc
     3781 tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag
     3841 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg
     3901 ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     3961 gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt
     4021 ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgg
     4081 ttctttccgc ctcagaaggt acctaaccaa gttcctcttt cagaggttat ttcaggccat
     4141 ggtgctgcgc agatccgcgt atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     4201 accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgatttta ccacatttgt
     4261 agaggtttta cttgcttaaa aaaacctccc acatctcccc ctgaacctga aacataaaat
     4321 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     4381 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4441 caaactcatc aatgtatctt atcatgtctg ctcgaagcgg ccaaggggtt atgctagtta
     4501 ttgctcagcg gtggcagcag ccaactcagc aagcggccgc aaaatcctgc aggaattggg
     4561 cccaaatcta gattattagt cactgtcatc cccagtctcc tccttttctt cctgagccat
     4621 ggagtcttct ccctcttccc tcagtcctgc gaagaagtca tcggtgctcc aagtcttgct
     4681 gctcagagcc cgcagtggtc ctcccttttt tttgcgccga cggtagtcat ccaccaccac
     4741 agctcgcagc tgggccatgt cccaaaactt gaggcgctgg tcatggccac tactggccag
     4801 gaagcggcca cagtgggaga gggccagctc ctccacaggc tccccagtgt gctggcccac
     4861 actgcccacc actcggttcg gtaggatgtt cacagccctg atgactccat cagtggagcc
     4921 agtacacagc agactctcgg tgactggaac catgcagtcg atagattcag ctctcagggc
     4981 aaagcggtca cttgtggccc caaagccatt ccaattgaag aggtagatgg taccttcact
     5041 ggagccacag gctaccttct tcccccattt catgagagtg acagaggtca ggtccccaga
     5101 ctgaggttct gagagcagct caaaccgacg cctcttaatg ttgaagatgc caaggcagcc
     5161 atccccgctg gctgtcagca gcagcttttt ggctggatcc agagccatgt ctgcgatgta
     5221 ctcttcatgt tgcctcatat ccattaaggg gccctccttc cgctggtccc agagacggat
     5281 accacctgtg tcatccccag tggccagaac attctcatcc accagcagaa gactgttgat
     5341 gggggcacca tgagccttgg aaacacgtct ttccagttgg ccctgctcca catctagaac
     5401 atggatggct ttgtccttgg agacagtaat gagcttctgc ccatcttcag agaaggccac
     5461 agctcggcag gccttgagat ggtgacctga tgaccagagc tccttggttt ctccctcttg
     5521 gcaagagtag gaaaagacga acacgtcccc gtccacgtcc cctgcagcca gtaggtcacg
     5581 ggccggatgg aacgccagcc cactagccgg agcttccagc acgatgtctt ccggagtgtc
     5641 ccggatccgg gttggggctt ccatggagtc tgggtcctcc tcgtcgctcc catcctcagc
     5701 gggcctctcc tcacaagtgc ggtccatgaa ttcggaagcg atcgcgccgc tcgagcccgc
     5761 cagaatgcgt tcgcacagcc gccagccggt cactccgttg atggttactc ggaacagcag
     5821 ggagccgtcg gggttgatca ggcgctcgtc gataattttg ttgccgttcc acagggtccc
     5881 tgttacagtg atctttttgc cgtcgaacac ggcgatgcct tcatacggcc gtccgaaata
     5941 gtcgatcatg ttcggcgtaa ccccgtcgat taccagtgtg ccatagtgca ggatcacctt
     6001 aaagtgatga tcatccacag ggtacaccac cttaaaaatt ttttcgatct ggcccatttg
     6061 gtcgccgctc agaccttcat acgggatgat gacatggatg tcgatcttca gcccattttc
     6121 accgctcagg acaatccttt ggatcggagt tacggacacc ccgagattct gaaacaaact
     6181 ggacacacct ccctgttcaa ggacttggtc caggttgtag ccggctgtct gtcgccagtc
     6241 cccaacgaaa tcttcgagtg tgaagaccat ggtgagcgct agccctatag tgagtcgtat
     6301 taatactcta gccttaagag ctgtaattga actgggagtg gacacctgtg gagagaaagg
     6361 caaagtggat gtcagtaaga ccaataggtg cctatcagaa acgcaagagt cttctctgtc
     6421 tcgacaagcc cagtttctat tggtctcctt aaacctgtct tgtaaccttg atacttacct
     6481 gcccagtgcc tcacgaccaa cttctgcagc ttaagttcga gactgttgtg tcagaagcac
     6541 tgactgcgtt agcaatttaa ctgtgataaa ctaccgcaat aaagcttcta gtgatctgac
     6601 ggttcactaa accagctctg cttatataga cctcccaccg tacacgccta ccgcccattt
     6661 gcgtcaacgg ggcggggtta ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
     6721 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct
     6781 atccacgccc attggtgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata
     6841 cgtagatgta ctgccaagta ggaaagtccc gtaaggtcat gtactgggca taatgccagg
     6901 cgggccattt accgtcattg acgtcaatag ggggcggact tggcatatga tacacttgat
     6961 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     7021 ctattggcgt tactatggga acatacgtca ttattg
//
