LOCUS       sequence-425789-        7467 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of full-length human FBXW10 isoform 2. 
            N-terminal HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7467)
  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 7467)
  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: pBiT3.1-N_FBXW10:1-1061
FEATURES             Location/Qualifiers
     source          1..7467
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    24..105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        338..641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        642..845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     796..816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          981..1113
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1096..1115
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1157..1176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1157..1175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2261..2270
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4530..4651
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4541..4560)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4766..4799)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4809..4856
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4888..5682
                     /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(4942..4961)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5552..5571
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5840..6428)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5920..5939)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6554..6602
                     /note="synthetic polyadenylation signal"
     CDS             complement(6626..7024)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      7021..7030
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7055..7412)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7069..7204
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        7221..7412
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7369..7389
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagacgcgtt aattaagacg tcggcagtga aaaaaatgct ttatttgtga aatttgtgat
       61 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
      121 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
      181 ctctacaaat gtggtatggc tgattatgat ccgcggccgc aatacgtcga cgttatcagc
      241 tgacttcgta cgagagccta ggattatggc gcgccactag ttattaatag taatcaatta
      301 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      361 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      421 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      481 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
      541 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
      601 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      661 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      721 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
      781 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      841 gagctggttt agtgaaccgt cagatcacta gaatctttat tgcggtagtt tatcacagtt
      901 aaattgctaa cgcagtcagt gcttctgaca caacagtctc gaacttaagc tgcagaagtt
      961 ggtcgtgagg cactgggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat
     1021 agaaactggg cttgtcgaga cagagaagac tcttgcgttt ctgataggca cctattggtc
     1081 ttactgacat ccactttgcc tttctctcca caggtgtcca ctcccagttc aattacagct
     1141 cttaaggcta gagtattaat acgactcact ataggggtta gcgatcgcca tggtgagcgg
     1201 ctggcggctg ttcaagaaga ttagcgggag ctccggtggc tcgagcggta tggaaaacct
     1261 ggaatcaagg ctcaagaatg ccccctattt tcgttgtgag aagggaaccg attccatccc
     1321 tctatgccgg aagtgtgaga cgtgtgtctt agcctggaag atcttctcta ccaaagagtg
     1381 gttctgcagg atcaatgaca tatcacagag gaggtttcta gttggcattc tgaagcagtt
     1441 aaatagctta tatttgttac actatttcca aaatatcctt cagaccacac agggaaagga
     1501 tttcatctat aacaggtccc ggatcaacct cagcaagaaa gaggggaaag ttgtgaagtc
     1561 ctccttgaac caaatgttgg ataaaacagt agaacagaag atgaaagaga tcttgtactg
     1621 gtttgcgaac agcacccagt ggaccaaggc gaattatact ctcttactgc tgcagatgtg
     1681 caaccccaaa ttactgctca ctgctgccaa tgtgatcaga gtcctgtttc tgagagagga
     1741 gaacaatatc tcagggctca atcaagacat cacagatgtg tgtttttccc ctgagaaaga
     1801 ccacagctcc aagtctgcga cctcacaagt ctattggaca gccaaaactc agcacacatc
     1861 ccttcctttg tccaaagccc cagaaaatga acacttgctt ggggcagcat ctaaccctga
     1921 ggaaccatgg aggaattcac tccggtgtat atccgaaatg aataggctgt tttctggaaa
     1981 aggagacata accaagccag ggtacgatcc ctgcaatcta ttggttgacc tggatgacat
     2041 cagagacctg tcttctgggt tcagcaaata cctagacttc atccgttacc tgcccatcca
     2101 cctctccaag tacattctaa gtatgctggg gtgtatgagg ggatttccca gacacctccc
     2161 atcttggccc agttctgggg tttctgggtc catctttttt tttccaggaa tgctggatag
     2221 acacaccctg aacaagtgcg cctctgtgag ccagcactgg gccgccatgg ctcaacaggt
     2281 caagatggac ttgtcagcgc acggcttcat tcagaaccag attaccttct tgcaggggtc
     2341 ctacacaaga ggaattgatc ctaattatgc caataaggtt tctatcccag ttcctaaaat
     2401 ggtagatgac gggaagagca tgcgtgtgaa acatccgaag tggaagctga gaacgaagaa
     2461 tgagtacaac ctgtggactg cataccagaa cgaggaaacg cagcaggtcc tgatagagga
     2521 gagaaatgtt ttctgtggga cctacaatgt tcgcattctc tctgacacgt gggatcaaaa
     2581 ccgagtcatc cactattccg ggggagatct gatagctgtg tcatctaatc gaaagatcca
     2641 tcttctggac atcatacaag tgaaagcgat acccgttgaa ttccgaggcc atgctgggag
     2701 tgtccgggcc ctcttcctgt gtgaggagga aaactttctc ctaagcggga gctatgacct
     2761 aagtatcaga tactgggatc tgaaaagtgg ggtttgcaca cgaatcttcg gtggtcacca
     2821 ggggactatc acttgcatgg acttgtgtaa gaacaggctc gtatctggag gaagagattg
     2881 tcaggtaaaa gtatgggatg tagacacagg gaagtgcctg aagacgttta gacacaaaga
     2941 ccccatcttg gccaccagga tcaatgatac ctacattgtg agcagctgtg agcgagggct
     3001 ggtgaaagtg tggcacattg ccatggccca gttggtaaag actctcagtg gccatgaggg
     3061 agccgtgaaa tgcctgttct ttgaccagtg gcatctcctc tcaggaagta ctgatggcct
     3121 ggtcatggcc tggagcatgg tggggaagta cgagcgctgc ctgatggcct tcaagcatcc
     3181 caaggaggtg ctcgacgtgt ccctcctctt cctccgggtc atcagcgcct gtgcagatgg
     3241 caagatccga atttacaatt tcctcaacgg gaactgtatg aaggtgttaa aagccaatgg
     3301 cagaggtgat cctgtgctgt ccttctttat tcagggcaac aggatggtgg tcaacacaga
     3361 gagcaatgtt ctcatgttcc agtttgagca cataaagtgg cagtatgccg tggaaaaaac
     3421 gaaacaaaag aagaataagg agaaagagga ggaaaaagaa gaaaatagtc tcatggaaat
     3481 tctctctaag tgtaatattc aggttcacag cccaagagag tctgtatcca gtaaacaaac
     3541 tgtgatccaa gagctcctac caggcaaacc tcccaagtcc cgagtactcc tgaagccggc
     3601 caagttctct tcagatgatg tggagaaagc acaaaaacaa ggacaattgg aaactcctgg
     3661 aaaactgccc agtcacccaa agaaaaagtc ttggaaaatc cctatgtcac ctgaccaatt
     3721 cctcctgact gttagcgccc tgcagcacgc ccataattcc ggggaatttg cctatccctg
     3781 taggccccaa acagaaatta ctgatgtctg gggaccttca atttcatacc caaggaaggt
     3841 cttgaatttc aaaggaaaat caatccaacg tgcagttgat cggttgagat tgagcaatcc
     3901 tcctatagat gtgaaacgaa ccagtattcc ccttgaaatc cagaaactgc agcccaactt
     3961 gaagatctct ttgcacagtc ctagagtcca gtccaccata ccccagccca tgattatccg
     4021 ctccaggttc tctggcagct taaagggtgg agaccaagtg accagttcaa ttgaaagggc
     4081 tgtgtgcagt acgggtcccc tgaccagtat gcaggtcatt aaaccaaacc gcatgctagc
     4141 tccacaagtg ggcacagcca ccctgtctct taagaaagaa cggcctcgca tctatacagc
     4201 ccttgatcct tttagagtga acactgagtt cgtgctgttg accgtgaagg aggagaagga
     4261 gcaccaggaa gccaagatga aggaatatca ggccagggag tccactggag tggttgatcc
     4321 aggaaaagtc agcaaagctg catggatcag gaagatcaaa ggcctgccta ttgataattt
     4381 cacgaagcaa gggaaaacag cggcccctga acttggacaa aatgtattta tctaatctag
     4441 aggtcaagct tcgggggttt aaacctgcag gcatgcaagc tgatttggct gctgccaccg
     4501 ctgagcaagg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     4561 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     4621 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     4681 tcaggttcag ggggagatgt gggaggtttt tttaagcaag taaaacctct acaaatgtgg
     4741 taaaatctta attaagactt ctaatataac ttcgtatagc atacattata cgaagttatg
     4801 gttccactgt tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg
     4861 tgaggaacta aacccaggag gcagatcatg attgaacaag atggattgca cgcaggttct
     4921 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4981 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     5041 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     5101 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     5161 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     5221 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     5281 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     5341 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     5401 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5461 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5521 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5581 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5641 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gattcgaaat gaccgaccaa
     5701 gcgacgccca accggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     5761 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     5821 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5881 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5941 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6001 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6061 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6121 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6181 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6241 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     6301 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6361 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6421 gatttcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6481 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttat
     6541 agtccggaat cgtcacacaa aaaaccaaca cacagatgta atgaaaataa agatatttta
     6601 ttgcggccat cgtgatggct agagtttagc cctcccacac ataaccagag ggcagcaatt
     6661 cacgaatccc aactgccgtc ggctgtccat cactgtcctt cactatggct ttgatcccag
     6721 gatgcagatc gagaagcacc tgtcggcacc gtccgcaggg gctcaagatg cccctgttct
     6781 catttccgat cgcgacgata caagtcaggt tgccagctgc cgcagcagca gcagtgccca
     6841 gcaccacgag ttctgcacaa ggtcccccag taaaatgata tacattgaca ccagtgaaga
     6901 tgcggccgtc gctagagaga gctgcgctgg cgacgctgta gtcttcagag atggggatgc
     6961 tgttgattgt agccgttgct ctttcaatga gggtggattc ttcttgagac aaaggcttgg
     7021 ccatggtggc gctagtgtca gaagaatcga gctttttgca aaagcctagg cctccaaaaa
     7081 agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa
     7141 taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg
     7201 ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca
     7261 tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac taattgagat
     7321 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac
     7381 acacattcca cagctggttc tttccgcctc agaaggtacc taaccaagtt cctctttcag
     7441 aggttatttc aggccatggt gctgcgc
//
