LOCUS       sequence-425960-        5781 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of full-length human CDC20. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5781)
  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 5781)
  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_CDC20:1-499
FEATURES             Location/Qualifiers
     source          1..5781
                     /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    2844..2965
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2855..2874)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3080..3113)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3123..3170
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3202..3996
                     /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(3256..3275)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3866..3885
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4154..4742)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4234..4253)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4868..4916
                     /note="synthetic polyadenylation signal"
     CDS             complement(4940..5338)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5335..5344
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5369..5726)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5383..5518
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5535..5726
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5683..5703
                     /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 tggcacagtt
     1261 cgcgttcgag agtgacctgc actcgctgct tcagctggat gcacccatcc ccaatgcacc
     1321 ccctgcgcgc tggcagcgca aagccaagga agccgcaggc ccggccccct cacccatgcg
     1381 ggccgccaac cgatcccaca gcgccggcag gactccgggc cgaactcctg gcaaatccag
     1441 ttccaaggtt cagaccactc ctagcaaacc tggcggtgac cgctatatcc cccatcgcag
     1501 tgctgcccag atggaggtgg ccagcttcct cctgagcaag gagaaccagc ctgaaaacag
     1561 ccagacgccc accaagaagg aacatcagaa agcctgggct ttgaacctga acggttttga
     1621 tgtagaggaa gccaagatcc ttcggctcag tggaaaacca caaaatgcgc cagagggtta
     1681 ccagaacaga ctgaaagtac tctacagcca aaaggccact cctggctcca gccggaagac
     1741 ctgccgttac attccttccc tgccagaccg tatcctggat gcgcctgaaa tccgaaatga
     1801 ctattacctg aaccttgtgg attggagttc tgggaatgta ctggccgtgg cactggacaa
     1861 cagtgtgtac ctgtggagtg caagctctgg tgacatcctg cagcttttgc aaatggagca
     1921 gcctggggaa tatatatcct ctgtggcctg gatcaaagag ggcaactact tggctgtggg
     1981 caccagcagt gctgaggtgc agctatggga tgtgcagcag cagaaacggc ttcgaaatat
     2041 gaccagtcac tctgcccgag tgggctccct aagctggaac agctatatcc tgtccagtgg
     2101 ttcacgttct ggccacatcc accaccatga tgttcgggta gcagaacacc atgtggccac
     2161 actgagtggc cacagccagg aagtgtgtgg gctgcgctgg gccccagatg gacgacattt
     2221 ggccagtggt ggtaatgata acttggtcaa tgtgtggcct agtgctcctg gagagggtgg
     2281 ctgggttcct ctgcagacat tcacccagca tcaaggggct gtcaaggccg tagcatggtg
     2341 tccctggcag tccaatgtcc tggcaacagg agggggcacc agtgatcgac acattcgcat
     2401 ctggaatgtg tgctctgggg cctgtctgag tgccgtggat gcccattccc aggtgtgctc
     2461 catcctctgg tctccccatt acaaggagct catctcaggc catggctttg cacagaacca
     2521 gctagttatt tggaagtacc caaccatggc caaggtggct gaactcaaag gtcacacatc
     2581 ccgggtcctg agtctgacca tgagcccaga tggggccaca gtggcatccg cagcagcaga
     2641 tgagaccctg aggctatggc gctgttttga gttggaccct gcgcggcggc gggagcggga
     2701 gaaggccagt gcagccaaaa gcagcctcat ccaccaaggc atccgctaat ctagaggtca
     2761 agcttcgggg gtttaaacct gcaggcatgc aagctgattt ggctgctgcc accgctgagc
     2821 aaggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     2881 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     2941 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3001 tcagggggag atgtgggagg tttttttaag caagtaaaac ctctacaaat gtggtaaaat
     3061 cttaattaag acttctaata taacttcgta tagcatacat tatacgaagt tatggttcca
     3121 ctgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
     3181 actaaaccca ggaggcagat catgattgaa caagatggat tgcacgcagg ttctccggcc
     3241 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3301 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     3361 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     3421 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     3481 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     3541 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     3601 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     3661 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     3721 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     3781 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     3841 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     3901 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     3961 atcgccttct atcgccttct tgacgagttc ttctgattcg aaatgaccga ccaagcgacg
     4021 cccaaccggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4081 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4141 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4201 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4261 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4321 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4381 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4441 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4501 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4561 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     4621 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4681 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc
     4741 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4801 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttatagtccg
     4861 gaatcgtcac acaaaaaacc aacacacaga tgtaatgaaa ataaagatat tttattgcgg
     4921 ccatcgtgat ggctagagtt tagccctccc acacataacc agagggcagc aattcacgaa
     4981 tcccaactgc cgtcggctgt ccatcactgt ccttcactat ggctttgatc ccaggatgca
     5041 gatcgagaag cacctgtcgg caccgtccgc aggggctcaa gatgcccctg ttctcatttc
     5101 cgatcgcgac gatacaagtc aggttgccag ctgccgcagc agcagcagtg cccagcacca
     5161 cgagttctgc acaaggtccc ccagtaaaat gatatacatt gacaccagtg aagatgcggc
     5221 cgtcgctaga gagagctgcg ctggcgacgc tgtagtcttc agagatgggg atgctgttga
     5281 ttgtagccgt tgctctttca atgagggtgg attcttcttg agacaaaggc ttggccatgg
     5341 tggcgctagt gtcagaagaa tcgagctttt tgcaaaagcc taggcctcca aaaaagcctc
     5401 ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa
     5461 aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg
     5521 gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc
     5581 tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc
     5641 tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat
     5701 tccacagctg gttctttccg cctcagaagg tacctaacca agttcctctt tcagaggtta
     5761 tttcaggcca tggtgctgcg c
//
