LOCUS       sequence-427147-        8143 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of V5 tagged Drosophila Wdr20.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8143)
  AUTHORS   Spencer ZT, Ng VH, Benchabane H, Siddiqui GS, Duwadi D, Maines B, 
            Bryant JM, Schwarzkopf A, Yuan K, Kassel SN, Mishra A, Pimentel A, 
            Lebensohn AM, Rohatgi R, Gerber SA, Robbins DJ, Lee E, Ahmed Y
  TITLE     The USP46 deubiquitylase complex increases Wingless/Wnt signaling 
            strength by stabilizing Arrow/LRP6.
  JOURNAL   Nat Commun. 2023 Oct 5;14(1):6174. doi: 10.1038/s41467-023-41843-0.
  PUBMED    37798281
REFERENCE   2  (bases 1 to 8143)
  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):6174. doi"; pages: " 
            10.1038/s41467-023-41843-0"
COMMENT     Sequence Label: Drosophila Wdr20-V5
FEATURES             Location/Qualifiers
     source          1..8143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1113..1122
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1144..1163)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1145..1163)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1208..1411)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1237..1257)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1412..1791
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1963..1982
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2057..2161
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2162..2230,2231..3022)
                     /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(2380..2399)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3193..3781
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3682..3701
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3935..3952
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4069..4090
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4105..4110,4111..4128,4129..4135)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4143..4159
                     /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     4148..4170
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4167..4183
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4167..4183
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4232..4353
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4243..4262)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4297..4316
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4527..5321)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4638..4657)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5248..5267
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(5388..5717)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5402..5537
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5702..5722
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(5731..6159)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5841..5862)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6053..6072
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    6205..6429
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6418..6435
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(6476..6517)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
ORIGIN
        1 gcacacgcac aggaagccgc cgaagtagga tctggcgatg cccagcagct ccattgtatc
       61 gtagtggaac actctcagga agccatcctg gctcaccacg gccaggtggc tgccacaggg
      121 ggagaagcag aactcattca cacagcaatt atctgtgctg aacacccact tgaacagggg
      181 attcctggtg gtcttgctct tggatgtcag gatggtgtag ccatcgccca gcttgaaggg
      241 ctggtaagaa ggggctgtgg cggcgcaggg cagctcctca ttgtacaggt acaggtgtcc
      301 gctggcgtgg gcggccagga acaggtgagg gctgttgggc agccacttca ggcatgtcac
      361 ctttgtcttg tcgatcagtc tctcctcgtt gaacagcttc ctcacctctc tggggcccac
      421 gtgagggctc accagctgga tctggccggt tgtgaagcca accagcagag gggcgcctgt
      481 gggggtggcg ctgctgatgt tgaagtcgtg acaggaagga tttgtgccct tgtagaactt
      541 cttatcgata ggcttgctca tctcggtgcc cttcttggcg cctctgaagc tgtacacgta
      601 caggtctctg ccgaagttga agcagattct atcgccgccc agcacctgat ttgtggccac
      661 gccgttctga tccaccacgc cgccgttcat catggagtgc atgctgattc cgccgcccag
      721 cctggcctcc acggtgttgt tgccggcgct gttgctgttg tgattagagt ggctgtagtc
      781 tccgccggcg ccgccattgc tgatggcggt ggaggctcca gcagctcctg taggggaggc
      841 gccgggggag ctgccgttgg ttgtggtggt gttggctccg gctgttcctc cgcccacggg
      901 tgtgcccacg cccacatcgc tgcccagctt gccctgggca ggggaaggca gtgtcaccat
      961 gctcaccctc acctgggggc tgctctggtt ggagctgtag cccaccctgt taggtctgct
     1021 gtactcggac agtgtcagca gtctgtatgt gccctctctg gtcacgaact gggtcttcag
     1081 atcgtccttc acgctggcgt ccagctggtt ggccatggtg gcggcgctag ccagcttggg
     1141 tctccctata gtgagtcgta ttaatttcga taagccagta agcagtgggt tctctagtta
     1201 gccagagagc tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca
     1261 atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac
     1321 tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac
     1381 gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact aatacgtaga
     1441 tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc
     1501 atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg
     1561 ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg
     1621 gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc
     1681 agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa
     1741 ctaatgaccc cgtaattgat tactattaat aactagtcaa taatcaatgt caacgcgtat
     1801 atctggcccg tacatcgcga agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc
     1861 aattgtcggt caagccttgc cttgttgtag cttaaatttt gctcgcgcac tactcagcga
     1921 cctccaacac acaagcaggg agcagatact ggcttaacta tgcggcatca gagcagattg
     1981 tactgagagt gcaccatagg ggatcgggag atctcccgat ccgtcgacgt caggtggcac
     2041 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2101 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2161 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     2221 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     2281 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     2341 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     2401 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     2461 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     2521 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     2581 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     2641 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     2701 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     2761 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     2821 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     2881 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     2941 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3001 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     3061 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3121 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3181 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3241 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3301 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3361 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3421 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3481 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3541 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3601 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3661 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3721 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3781 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3841 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     3901 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     3961 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     4021 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     4081 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     4141 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
     4201 ctagctagag gtcgacggta tacagacatg ataagataca ttgatgagtt tggacaaacc
     4261 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     4321 tttgtaacca ttataagctg caataaacaa gttggggtgg gcgaagaact ccagcatgag
     4381 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
     4441 ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc
     4501 ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg
     4561 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
     4621 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc
     4681 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
     4741 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gcgcgccttg
     4801 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
     4861 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
     4921 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
     4981 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
     5041 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
     5101 cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg
     5161 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
     5221 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
     5281 gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct
     5341 gtctcttgat cagatccgaa aatggatata caagctcccg ggagcttttt gcaaaagcct
     5401 aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggcaga ggcggcctcg
     5461 gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc
     5521 ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga
     5581 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct
     5641 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
     5701 caccctaact gacacacatt ccacagaatt aattcgcgtt aaatttttgt taaatcagct
     5761 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     5821 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     5881 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     5941 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     6001 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     6061 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     6121 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtg gggatacccc ctagagcccc
     6181 agctggttct ttccgcctca gaagccatag agcccaccgc atccccagca tgcctgctat
     6241 tgtcttccca atcctccccc ttgctgtcct gccccacccc accccccaga atagaatgac
     6301 acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca gtgggagtgg
     6361 caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc tggcaactag
     6421 aaggcacagt cgaggctgat cagcgggttt aaacgggccc tctagactcg agtcatgtgc
     6481 tatccaggcc cagcagaggg ttagggattg gcttgccgct tccgccgccg ccgatcactg
     6541 ttcctccagg agcagcggcc tggccggggc tcaggtgctg tgtagcgtgg gtggcgtggc
     6601 cgggtctggc ccatgtgtag atgaagccat cctgacaggc tgtcaggaag cagtcctctc
     6661 tgaagatcag ggcggtcagc ctctcgtggg cgatcttctt gcacaccagg ggctccagca
     6721 gaggacactc gtcgaacctg ggacaggcgg gtgtgccgat cagctgcatg ggatcgaagc
     6781 tgctcactcc ggagccgctt ccggcagcgc ttccggaagc gtgagcggcg gtggccaggc
     6841 tggagtggtg ggaggacctg gacagcttca gggagccgaa ggagtgggct gtgctggagc
     6901 tgccgatagc gccggcggcg ttgctgtcca gggtgctgga gtggccgaag ttgctggagg
     6961 aggagttgtt ggactggctg tggttattgt ggccgcctgt gtggttgtgc tggttgtagg
     7021 acttcagcat gctcatggcc tttctgtgct gtctgtgggc tgtggagtgg ctgccctcgt
     7081 agccggcgcc cttcttctcg gagctgctca ggaaggagaa gttgctcagt ctctgggtca
     7141 ggctgttgaa ggcgctgaag ccggagccgc tattggagct cttgttgctg gagttggagg
     7201 tcttgctgtt gtagccgctt gtggtctggc taggtctttg tccagatccg gagccgccgc
     7261 cgccgtcgat ggaatcgctc tttgtggcgc tgatacagtt agggaacttg atgctgttgc
     7321 cggtgccgtg gtttctgtta ttctgcttgg tggaggcaac agctccagct ccggccacgc
     7381 tgccggcgcc ggcattagag gaggtgctgg cgggtgtggc ggcctctgtg tcgcaatcgt
     7441 cgggggagct ctgggaggag atggtacagt tggctgtgga gaacttggag ctggagctgt
     7501 tggagtgctc tccagcgccg ttgccggttc ctccgccagc agcctccctg gtggggctgc
     7561 aggagccgga ttcggcctgc tggcctgtgg cctggccgct catggccaca ggtctgatca
     7621 ccttgatgcc atcctcggcc tcggcctcct cgtccacgcc gccattcagg taggccctct
     7681 cgctattggc gggctgcctc agcaccagag ggtgcctcag cacgtcctct gtgatatccc
     7741 acaggcagat ctgggtgtcc tggctcacgc tgcccagcct gtagctcatc agcttctcgg
     7801 cgctgctggc atcgcttctg aaggaggcgc tgtgagggcc attcctgtcg gcgtgcacgg
     7861 gtgtgctgtt tctatcgaag ccctcgaagc cgccattggc tgtggaatcg ccggagaatc
     7921 tggcctctct gctgtggctg tactcgttca tctggttctc gtcatcgctg aagtcgccgc
     7981 catcccagtt tgtgtaggat gtggtgtagg ggtcgaaggc caccacgctc acccagcttc
     8041 tgtggccctg gcccctggcc accactcttc tctcgtgcag ggaccacact gtcaccaggt
     8101 cgtcctcgcc gcccaccacg atgtacttgc cgtcagggga cca
//
