LOCUS       sequence-427146-        6688 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of V5 tagged Drosophila Usp46.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6688)
  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 6688)
  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 Usp46-V5
FEATURES             Location/Qualifiers
     source          1..6688
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      385..394
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(416..435)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(417..435)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(480..683)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(509..529)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        684..1063
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1235..1254
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1329..1433
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1434..1502,1503..2294)
                     /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(1652..1671)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2465..3053
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2954..2973
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3207..3224
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3341..3362
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3377..3382,3383..3400,3401..3407)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3415..3431
                     /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     3420..3442
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3439..3455
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3439..3455
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3504..3625
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3515..3534)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3569..3588
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(3799..4593)
                     /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(3910..3929)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4520..4539
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4660..4989)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4674..4809
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4728..4747)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     4974..4994
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(5003..5431)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5113..5134)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5325..5344
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    5477..5701
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     5690..5707
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(5748..5789)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
ORIGIN
        1 tctcggccag gatgatctcg ttgatgtgat tgatcaggaa gttcaggaac tcgtgggcgt
       61 cctgctgcat gtaattgtcg aactcctcct tctcctttct cagccttgtg atgaacttct
      121 tgggggcgat gctgcccacc ttcttcttct gggtggcgat gctgtagaac aggtcggcca
      181 gacagctcag cagtgtctcc ttgggtctct tgttcttggc cttgtactcc agcaccttct
      241 ccctgaaggg cttgcagaag tacagggcct gcagcacgga gttgctgtag caggtattgc
      301 cgaagttcac caggccgaag tagtgctcat tgggggggaa caggtcggag ccgatctccc
      361 tctccagctg ggacacgtta gcgcccatgg tggcggcgct agccagcttg ggtctcccta
      421 tagtgagtcg tattaatttc gataagccag taagcagtgg gttctctagt tagccagaga
      481 gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caatggggcg
      541 gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg
      601 acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg
      661 atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc
      721 caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg ccatttaccg
      781 tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac tgccaagtgg
      841 gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact
      901 atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc
      961 gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac
     1021 cccgtaattg attactatta ataactagtc aataatcaat gtcaacgcgt atatctggcc
     1081 cgtacatcgc gaagcagcgc aaaacgccta accctaagca gattcttcat gcaattgtcg
     1141 gtcaagcctt gccttgttgt agcttaaatt ttgctcgcgc actactcagc gacctccaac
     1201 acacaagcag ggagcagata ctggcttaac tatgcggcat cagagcagat tgtactgaga
     1261 gtgcaccata ggggatcggg agatctcccg atccgtcgac gtcaggtggc acttttcggg
     1321 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     1381 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     1441 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     1501 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1561 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1621 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     1681 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1741 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1801 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1861 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1921 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1981 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     2041 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     2101 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     2161 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     2221 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     2281 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     2341 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     2401 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2461 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2521 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2581 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     2641 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2701 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2761 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2821 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2881 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2941 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3001 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3061 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     3121 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     3181 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     3241 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     3301 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     3361 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     3421 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag ctctagctag
     3481 aggtcgacgg tatacagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3541 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3601 cattataagc tgcaataaac aagttggggt gggcgaagaa ctccagcatg agatccccgc
     3661 gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga
     3721 aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt
     3781 cgaaccccag agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg
     3841 cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag
     3901 ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag
     3961 ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca
     4021 ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc
     4081 gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag
     4141 accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg
     4201 gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt
     4261 ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag
     4321 ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt
     4381 ggccagccac gatagccgcg ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc
     4441 ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga
     4501 gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg
     4561 agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg
     4621 atcagatccg aaaatggata tacaagctcc cgggagcttt ttgcaaaagc ctaggcctcc
     4681 aaaaaagcct cctcactact tctggaatag ctcagaggca gaggcggcct cggcctctgc
     4741 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     4801 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct
     4861 ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt
     4921 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
     4981 ctgacacaca ttccacagaa ttaattcgcg ttaaattttt gttaaatcag ctcatttttt
     5041 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     5101 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     5161 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     5221 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     5281 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     5341 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     5401 gccgcgctta atgcgccgct acagggcgcg tggggatacc ccctagagcc ccagctggtt
     5461 ctttccgcct cagaagccat agagcccacc gcatccccag catgcctgct attgtcttcc
     5521 caatcctccc ccttgctgtc ctgccccacc ccacccccca gaatagaatg acacctactc
     5581 agacaatgcg atgcaatttc ctcattttat taggaaagga cagtgggagt ggcaccttcc
     5641 agggtcaagg aaggcacggg ggaggggcaa acaacagatg gctggcaact agaaggcaca
     5701 gtcgaggctg atcagcgggt ttaaacgggc cctctagact cgagtcaggt ggagtccagg
     5761 cccagcagag ggttggggat tggctttcca gatccgccgc cgccggcgca gtctctggac
     5821 tggtagaaca ggatgtagcc tgtctcgctg gacttgtgga tgtcgctggt caggccgtag
     5881 aagtcctcga tggtggaggc ctcgatcttg tccaccatgt cgtcgtcgaa cagcagccac
     5941 aggccgtggc tcttcacgat gctgatgtag tggcctctat ttgggccgga gccgcagtgg
     6001 atcaccacgg ctgtcagatc gtacagtcta tcagggttca cggcgtcgtc gctggtgttg
     6061 aacagtctca gctccagagg gaacaccacc ctgtgggaca ccttgatgtg tctgttgaac
     6121 tgctccatgt acttgaatct cttcaggtgc agggccagga tcataggcag cttcttcact
     6181 ctcattctct tctgggcctc ctggtagctg cagcagttat cacacttgaa cttattgtcg
     6241 ctgcacaggg tctcggtgtt gctgaagcac ctcaggcagt gggtgatgga ggtattctgg
     6301 tccacgtcca cctgcagatc gaagaagttc tcatccttgg agctcactgt ctcacagttc
     6361 aggcacctgg tctcgcttgt caggatgccc tggaagatct cgtgcaccca tgtaggctcg
     6421 ctattggcgc cattggtgcc ggtgccgttg ccggagatga taggtgtggt ggtggcggtc
     6481 agggagccgg aggcatccag cacggaggtg ttggcattca ggctgcctgt ggaattggag
     6541 gagttgccgt tgctgtttgt tgtgctattg gagttgctat tgctgttgct tgtgctggag
     6601 ctcttgctgg cgatggagga ggccatggcg cttgtgctgc cgccctggtt ggtggcctta
     6661 gggttgccat tggaggggcc ggcgtttc
//
