LOCUS       sequence-426018-        5364 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of full-length human WDR45. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5364)
  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 5364)
  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_WDR45:1-360
FEATURES             Location/Qualifiers
     source          1..5364
                     /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"
     polyA_signal    2427..2548
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2438..2457)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2663..2696)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2706..2753
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2785..3579
                     /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(2839..2858)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3449..3468
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3737..4325)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3817..3836)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4451..4499
                     /note="synthetic polyadenylation signal"
     CDS             complement(4523..4921)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4918..4927
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4952..5309)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4966..5101
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5118..5309
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5266..5286
                     /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 tgactcaaca
     1261 gccacttcga ggagtgacca gcctgcgttt caaccaagac caaagctgct tttgctgcgc
     1321 catggagaca ggtgtgcgca tctacaacgt ggagcccttg atggagaagg ggcatctgga
     1381 ccacgagcag gtgggcagca tgggcttggt ggagatgctg caccgctcca accttctggc
     1441 cttggtgggc ggtggtagta gtcccaagtt ctcagagatc tcagtgctga tctgggacga
     1501 tgcccgggag ggcaaggact ccaaggagaa gctggtgctg gagttcacct tcaccaagcc
     1561 agtgctttct gtgcgcatgc gccatgacaa gatcgtgatc gtgctgaaga accgcatcta
     1621 tgtgtactcc ttccccgaca atccccgaaa gctgtttgag tttgataccc gggacaaccc
     1681 caaggggctc tgtgacctct gccccagcct ggagaagcaa ctgctagtgt tcccgggaca
     1741 caagtgtggg agtctgcaac ttgtggacct ggcgagcaca aagcctggca cctcgtctgc
     1801 tccattcacg atcaatgcac atcagagtga catagcctgt gtgtctctaa accagccagg
     1861 cactgtagtg gcctcagcct cccagaaggg tacccttatt cgcctctttg acacacaatc
     1921 caaggagaaa ctggtggagc tgcgccgagg cactgaccct gccaccctct actgcattaa
     1981 cttcagccac gactcctcct tcctctgcgc ttccagtgat aagggtactg tccatatctt
     2041 tgctctcaag gatacccgcc tcaaccgccg ctccgcgctg gctcgcgtgg gcaaggtggg
     2101 gcctatgatt gggcagtacg tggactctca gtggagcctg gcgagcttca ctgtgcctgc
     2161 tgagtcagct tgcatctgcg ccttcggtcg caatacttcc aagaacgtca actctgtcat
     2221 tgccatctgc gtagatggga ccttccacaa atatgtcttc actcctgatg gaaactgcaa
     2281 cagagaggct ttcgacgtgt accttgacat ctgtgatgat gatgactttt aatctagagg
     2341 tcaagcttcg ggggtttaaa cctgcaggca tgcaagctga tttggctgct gccaccgctg
     2401 agcaaggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     2461 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     2521 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     2581 ggttcagggg gagatgtggg aggttttttt aagcaagtaa aacctctaca aatgtggtaa
     2641 aatcttaatt aagacttcta atataacttc gtatagcata cattatacga agttatggtt
     2701 ccactgttga caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga
     2761 ggaactaaac ccaggaggca gatcatgatt gaacaagatg gattgcacgc aggttctccg
     2821 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     2881 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     2941 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     3001 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3061 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3121 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3181 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3241 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3301 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3361 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3421 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3481 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3541 cgcatcgcct tctatcgcct tcttgacgag ttcttctgat tcgaaatgac cgaccaagcg
     3601 acgcccaacc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3661 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3721 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3781 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3841 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3901 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3961 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4021 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4081 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4141 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     4201 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4261 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4321 ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4381 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttatagt
     4441 ccggaatcgt cacacaaaaa accaacacac agatgtaatg aaaataaaga tattttattg
     4501 cggccatcgt gatggctaga gtttagccct cccacacata accagagggc agcaattcac
     4561 gaatcccaac tgccgtcggc tgtccatcac tgtccttcac tatggctttg atcccaggat
     4621 gcagatcgag aagcacctgt cggcaccgtc cgcaggggct caagatgccc ctgttctcat
     4681 ttccgatcgc gacgatacaa gtcaggttgc cagctgccgc agcagcagca gtgcccagca
     4741 ccacgagttc tgcacaaggt cccccagtaa aatgatatac attgacacca gtgaagatgc
     4801 ggccgtcgct agagagagct gcgctggcga cgctgtagtc ttcagagatg gggatgctgt
     4861 tgattgtagc cgttgctctt tcaatgaggg tggattcttc ttgagacaaa ggcttggcca
     4921 tggtggcgct agtgtcagaa gaatcgagct ttttgcaaaa gcctaggcct ccaaaaaagc
     4981 ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa
     5041 aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga
     5101 tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac
     5161 ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca
     5221 tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca
     5281 cattccacag ctggttcttt ccgcctcaga aggtacctaa ccaagttcct ctttcagagg
     5341 ttatttcagg ccatggtgct gcgc
//
