LOCUS       sequence-423449-        5432 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human RAE1. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5432)
  AUTHORS   Ackloo S, Li F, Szewczyk M, Seitova A, Loppnau P, Zeng H, Xu J, 
            Ahmad S, Arnautova YA, Baghaie AJ, Beldar S, Bolotokova A, Centrella
            PA, Chau I, Clark MA, Cuozzo JW, Dehghani-Tafti S, Disch JS, Dong A,
            Dumas A, Feng JA, Ghiabi P, Gibson E, Gilmer J, Goldman B, Green SR,
            Guie MA, Guilinger JP, Harms N, Herasymenko O, Houliston S, 
            Hutchinson A, Kearnes S, Keefe AD, Kimani SW, Kramer T, Kutera M, 
            Kwak HA, Lento C, Li Y, Liu J, Loup J, Machado RAC, Mulhern CJ, 
            Perveen S, Righetto GL, Riley P, Shrestha S, Sigel EA, Silva M, 
            Sintchak MD, Slakman BL, Taylor RD, Thompson J, Torng W, 
            Underkoffler C, von Rechenberg M, Walsh RT, Watson I, Wilson DJ, 
            Wolf E, Yadav M, Yazdi AK, Zhang J, Zhang Y, Santhakumar V, Edwards 
            AM, Barsyte-Lovejoy D, Schapira M, Brown PJ, Halabelian L, 
            Arrowsmith CH
  TITLE     A Target Class Ligandability Evaluation of WD40 Repeat-Containing 
            Proteins.
  JOURNAL   J Med Chem. 2025 Jan 23;68(2):1092-1112. doi: 
            10.1021/acs.jmedchem.4c02010. Epub 2024 Nov 4.
  PUBMED    39495097
REFERENCE   2  (bases 1 to 5432)
  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"; doi: 
            "10.1021/acs.jmedchem.4c02010"; journalName: "J Med Chem"; date: 
            "2025-01-23- 23"; volume: "68"; issue: "2"; pages: "1092-1112"
COMMENT     Sequence Label: pBiT3.1-C_RAE1:1-368
FEATURES             Location/Qualifiers
     source          1..5432
                     /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"
     CDS             1948..1959
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2495..2616
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2506..2525)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2731..2764)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2774..2821
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2853..3647
                     /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(2907..2926)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3517..3536
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3805..4393)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3885..3904)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4519..4567
                     /note="synthetic polyadenylation signal"
     CDS             complement(4591..4989)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4986..4995
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5020..5377)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5034..5169
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5186..5377
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5334..5354
                     /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 ataggggcga tcgcaacaag cttcgggaag
     1201 atctggagct agcggcgggg gatccggtgg gaattcgaca ccatgagcct gtttggaaca
     1261 acctcaggtt ttggaaccag tgggaccagc atgtttggca gtgcaactac agacaatcac
     1321 aatcccatga aggatattga agtaacatca tctcctgatg atagcattgg ttgtctgtct
     1381 tttagcccac caaccttgcc ggggaacttt cttattgcag gatcatgggc taatgatgtt
     1441 cgctgctggg aagttcaaga cagtggacag accattccaa aagcccagca gatgcacact
     1501 gggcctgtgc ttgatgtctg ctggagtgac gatgggagca aagtgtttac ggcatcgtgt
     1561 gataaaactg ccaaaatgtg ggacctcagc agtaaccaag cgatacagat cgcacagcat
     1621 gatgctcctg ttaaaaccat ccattggatc aaagctccaa actacagctg tgtgatgact
     1681 gggagctggg ataagacttt aaagttttgg gatactcgat cgtcaaatcc tatgatggtt
     1741 ttgcaactcc ctgaaaggtg ttactgtgct gacgtgatat accccatggc tgtggtggca
     1801 actgcagaga ggggcctgat tgtctatcag ctagagaatc aaccttctga attcaggagg
     1861 atagaatctc cactgaaaca tcagcatcgg tgtgtggcta tttttaaaga caaacagaac
     1921 aagcctactg gttttgccct gggaagtatc gaggggagag ttgctattca ctatatcaac
     1981 cccccgaacc ccgccaaaga taacttcacc tttaaatgtc atcgatctaa tggaaccaac
     2041 acttcagctc ctcaggacat ttatgcggta aatggaatcg cgttccatcc tgttcatggc
     2101 acccttgcaa ctgtgggatc tgatggtaga ttcagcttct gggacaaaga tgccagaaca
     2161 aaactaaaaa cttcggaaca gttagatcag cccatctcag cttgctgttt caatcacaat
     2221 ggaaacatat ttgcatacgc ttccagctac gactggtcaa agggacatga attttataat
     2281 ccccagaaaa aaaattacat tttcctgcgt aatgcagccg aagagctaaa gcccaggaat
     2341 aagaagggct cgagcggtgg gagctccggt gtgagcggct ggcggctgtt caagaagatt
     2401 agctaagttt aaacggccgc gactctagac tgcaggcatg caagctgatt tggctgctgc
     2461 caccgctgag caaggccgct tcgagcagac atgataagat acattgatga gtttggacaa
     2521 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     2581 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     2641 atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa
     2701 tgtggtaaaa tcttaattaa gacttctaat ataacttcgt atagcataca ttatacgaag
     2761 ttatggttcc actgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
     2821 caaggtgagg aactaaaccc aggaggcaga tcatgattga acaagatgga ttgcacgcag
     2881 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     2941 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3001 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     3061 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     3121 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     3181 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     3241 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     3301 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     3361 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     3421 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     3481 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     3541 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     3601 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgattc gaaatgaccg
     3661 accaagcgac gcccaaccgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3721 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3781 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3841 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3901 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3961 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4021 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4081 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4141 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4201 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     4261 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4321 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4381 aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4441 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4501 tttatagtcc ggaatcgtca cacaaaaaac caacacacag atgtaatgaa aataaagata
     4561 ttttattgcg gccatcgtga tggctagagt ttagccctcc cacacataac cagagggcag
     4621 caattcacga atcccaactg ccgtcggctg tccatcactg tccttcacta tggctttgat
     4681 cccaggatgc agatcgagaa gcacctgtcg gcaccgtccg caggggctca agatgcccct
     4741 gttctcattt ccgatcgcga cgatacaagt caggttgcca gctgccgcag cagcagcagt
     4801 gcccagcacc acgagttctg cacaaggtcc cccagtaaaa tgatatacat tgacaccagt
     4861 gaagatgcgg ccgtcgctag agagagctgc gctggcgacg ctgtagtctt cagagatggg
     4921 gatgctgttg attgtagccg ttgctctttc aatgagggtg gattcttctt gagacaaagg
     4981 cttggccatg gtggcgctag tgtcagaaga atcgagcttt ttgcaaaagc ctaggcctcc
     5041 aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc
     5101 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     5161 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct
     5221 ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt
     5281 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
     5341 ctgacacaca ttccacagct ggttctttcc gcctcagaag gtacctaacc aagttcctct
     5401 ttcagaggtt atttcaggcc atggtgctgc gc
//
