LOCUS       sequence-429330-        5956 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of FLAG tagged WDR20.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5956)
  AUTHORS   Ng VH, Spencer Z, Neitzel LR, Nayak A, Loberg MA, Shen C, Kassel SN,
            Kroh HK, An Z, Anthony CC, Bryant JM, Lawson A, Goldsmith L, 
            Benchabane H, Hansen AG, Li J, D'Souza S, Lebensohn AM, Rohatgi R, 
            Weiss WA, Weiss VL, Williams C, Hong CC, Robbins DJ, Ahmed Y, Lee E
  TITLE     The USP46 complex deubiquitylates LRP6 to promote Wnt/beta-catenin 
            signaling.
  JOURNAL   Nat Commun. 2023 Oct 5;14(1):6173. doi: 10.1038/s41467-023-41836-z.
  PUBMED    37798301
REFERENCE   2  (bases 1 to 5956)
  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: "2023-10-5"; volume: "14(1):6173. doi"; pages: " 
            10.1038/s41467-023-41836-z"
COMMENT     Sequence Label: Human FLAG-WDR20
FEATURES             Location/Qualifiers
     source          1..5956
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..158
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(61..80)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     115..134
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    203..227
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(1964..1987)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    complement(2034..2058)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(2113..2131)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2114..2131)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        complement(2166..3150)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     rep_origin      3221..3676
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3308..3327)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3518..3539
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3702..3806
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3807..3875,3876..4667)
                     /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(4025..4044)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4838..5426
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5327..5346
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5580..5597
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5714..5735
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5750..5755,5756..5773,5774..5780)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5788..5804
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5793..5815
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5812..5828
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5812..5828
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5847..5867
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5849..5867
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
       61 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
      121 tgtccaaact catcaatgta tcttatcatg tctggatcta cgtaatacga ctcactatag
      181 ttctagaggc tcgagaggat caaccacttt gtacaagaaa gctgggtcta gatatctcga
      241 gtgcggccgc gaattcttaa ggattaaaac ttaccacttt accaggcctt ccccatgtgc
      301 aaataaatcc ctcctgacaa gcagtgacta tacagtcttc aagaaatatt agtacagtca
      361 gtctctcatg tgctatcttt ttacatatca gcggctctaa caagggaaca tcttccattc
      421 gaggacacag gggcgttccc agagttttag cagggtccgt tttggttttg gtaactagat
      481 tcagtttgtc actgctcttg ctagaaatgt gtcccatgct atgatttcgc ttgtgatctt
      541 tctcgtggtg cctctccttc cggtcatgta gtgaaagtgt tgcaaatttg ctgaccccag
      601 aagcaatggc cccgtccatg acactgcttt tgctgccagc atttgagact gctgaatgtg
      661 gaaggctgtt ggaccgtggc agaggaggcg gcacagagtt cccgggtgtt gtaacactgt
      721 tcccattgct tccagcagga ggactcgtgg cattcatgac atttgtgtgt gtccttgctc
      781 ttgagagggg ttggtgaggg aaaaggatat cttctgtaag gtcccataaa cagagctgtg
      841 tgtcctggcc cacggaacca aaccgatacg tgacacttac ggggcggctg tctgtagagt
      901 tccgtttgga gagcctggac tgtgtactat ttgctcgatc tctgccaaaa tgaagaaggt
      961 cttggaagtc ctcatcgctg ccactaaact ccatagggtc accttcttct acactagtgg
     1021 tataagggtc aaacgctaca acactgaccc aggacttgtg cccgtggcct ctggctatta
     1081 ctcggcagtc tacaaaggac cagactgtca ccaagtcgtc ctccccacct gtcacgatgt
     1141 acttgccatc cgggctccag cacacacaca gcaagccccc aaagtagctt ttcatcgtac
     1201 cgtgcagctc cactgagtca aagttgaaca cccgcagaaa cccgtcctgg ctcacgcacg
     1261 ctaagaactt gccatctggg gagaaagcaa actcgttgag ggccccctcg cccaccgtcc
     1321 acttaaggag agggttcctc gtggatttgc tcttgcaagt gtgcacggca aagctctctc
     1381 cctgcttcag aagctggtag tggggggctg tggtgccaca agtgtgctcc acattatata
     1441 agtacatgtt cccactcgag tgggctacta ggaaaaggct ttccgaaccg tgaacccatt
     1501 tgacacaggt aactcgtgac ttgtctatta gtctttcctc attaaaaagt ttgctagttt
     1561 cttttttgat tgggtctata agctggactt ggcctgcgga aaagcccact aggagagaga
     1621 cactttctgc tgtggctgtt aggtggttga agtcatgaca agtaggctgt gttcctttgt
     1681 atatcctttt atctattggt ttactcaagt cagcagcctt gcggaccccc ttgtagatat
     1741 agaagtacag ctcccggccc acattgaagc agaggcggtc gccgttgcca gactggtcgt
     1801 tgaggtttac gaaggagacg cggacagggt tggatccctg cgagttgaag ggcacccggt
     1861 tgggccggct gtactccgag tgcggcagca gcttgtacag accttcccgg gtggtgaatt
     1921 gggtcttaat ctcgttcatc tccttccctc ctccctccgt cgccttatcg tcgtcatcct
     1981 tgtaatcgcc catggtggta ccggatccag tcgactgaat tggttccggc gccagcctgc
     2041 ttttttgtac aaacttgttg atccttgaat tcgaatcgat gggatcctgc aaaaagaaca
     2101 agtagcttgt attctatagt gtcacctaaa tcaagcttgc tccgacgtcc ccaggcagaa
     2161 tggcggttcc ctaaacgagc attgcttata tagacctccc attaggcacg cctaccgccc
     2221 atttacgtca atggaacgcc catttgcgtc attgcccctc cccattgacg tcaatgggga
     2281 tgtacttggc agccatcgcg ggccatttac cgccattgac gtcaatggga gtactgccaa
     2341 tgtaccctgg cgtacttcca atagtaatgt acttgccaag ttactattaa tagatattga
     2401 tgtactgcca agtgggccat ttaccgtcat tgacgtcaat agggggcgtg agaacggata
     2461 tgaatgggca atgagccatc ccattgacgt caatggtggg tggtcctatt gacgtcaatg
     2521 ggcattgagc caggcgggcc atttaccgta attgacgtca atgggggagg cgccatatac
     2581 gtcaatagga ccgcccatat gacgtcaata ggaaagacca tatatagaga ccattgacgt
     2641 caatggggga gtggctatgg gcggtattag gaagccccat atatggtata tgggaccgcc
     2701 cattgggagg ggctatctac gtcaatagga aaacccatat atggaatact atatggcata
     2761 gggccaatac atagtattga acctggccaa tagccatatt ggcatagggc catattggat
     2821 attgcctata tattgatcct ggcatatagc caatatggcc gccattattg gcaccatgcc
     2881 aattcaatat ggcggacttg gcacagggcc aagacccctc ctcaaacttg gcaccgtgcc
     2941 aagtagaccc ctccccagtt ggcacagtgc caggtccgcc atattgaatt ggcatggtgc
     3001 caagtccgcc atattgaatt ggctggggtc cagatccacc atattgaatt ggcacgagtc
     3061 catatacgcc atattgaatt ggcacaggtc cagatccacc atattgaatt ggcatgagtc
     3121 catatacgcc atattgaatt ggcggtcgac tggcgtaata gcgaagaggc ccgcaccgat
     3181 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
     3241 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3301 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3361 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3421 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3481 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3541 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3601 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     3661 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3721 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3781 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3841 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3901 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3961 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     4021 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4081 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4141 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4201 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     4261 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4321 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     4381 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     4441 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4501 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4561 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4621 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4681 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4741 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4801 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4861 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4921 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4981 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5041 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5101 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5161 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5221 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5281 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5341 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5401 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5461 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5521 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     5581 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     5641 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     5701 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     5761 tgctcccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     5821 gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg
     5881 gtaccgggcc caatgcattg gcgccgcggc cgcgaattaa aaaacctccc acacctcccc
     5941 ctgaacctga aacata
//
