LOCUS       sequence-425921-        5388 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human RAE1. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5388)
  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 5388)
  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-N_RAE1:1-368
FEATURES             Location/Qualifiers
     source          1..5388
                     /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             1955..1966
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2451..2572
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2462..2481)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2687..2720)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2730..2777
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2809..3603
                     /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(2863..2882)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3473..3492
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3761..4349)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3841..3860)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4475..4523
                     /note="synthetic polyadenylation signal"
     CDS             complement(4547..4945)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4942..4951
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4976..5333)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4990..5125
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5142..5333
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5290..5310
                     /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 tgagcctgtt
     1261 tggaacaacc tcaggttttg gaaccagtgg gaccagcatg tttggcagtg caactacaga
     1321 caatcacaat cccatgaagg atattgaagt aacatcatct cctgatgata gcattggttg
     1381 tctgtctttt agcccaccaa ccttgccggg gaactttctt attgcaggat catgggctaa
     1441 tgatgttcgc tgctgggaag ttcaagacag tggacagacc attccaaaag cccagcagat
     1501 gcacactggg cctgtgcttg atgtctgctg gagtgacgat gggagcaaag tgtttacggc
     1561 atcgtgtgat aaaactgcca aaatgtggga cctcagcagt aaccaagcga tacagatcgc
     1621 acagcatgat gctcctgtta aaaccatcca ttggatcaaa gctccaaact acagctgtgt
     1681 gatgactggg agctgggata agactttaaa gttttgggat actcgatcgt caaatcctat
     1741 gatggttttg caactccctg aaaggtgtta ctgtgctgac gtgatatacc ccatggctgt
     1801 ggtggcaact gcagagaggg gcctgattgt ctatcagcta gagaatcaac cttctgaatt
     1861 caggaggata gaatctccac tgaaacatca gcatcggtgt gtggctattt ttaaagacaa
     1921 acagaacaag cctactggtt ttgccctggg aagtatcgag gggagagttg ctattcacta
     1981 tatcaacccc ccgaaccccg ccaaagataa cttcaccttt aaatgtcatc gatctaatgg
     2041 aaccaacact tcagctcctc aggacattta tgcggtaaat ggaatcgcgt tccatcctgt
     2101 tcatggcacc cttgcaactg tgggatctga tggtagattc agcttctggg acaaagatgc
     2161 cagaacaaaa ctaaaaactt cggaacagtt agatcagccc atctcagctt gctgtttcaa
     2221 tcacaatgga aacatatttg catacgcttc cagctacgac tggtcaaagg gacatgaatt
     2281 ttataatccc cagaaaaaaa attacatttt cctgcgtaat gcagccgaag agctaaagcc
     2341 caggaataag aagtaatcta gaggtcaagc ttcgggggtt taaacctgca ggcatgcaag
     2401 ctgatttggc tgctgccacc gctgagcaag gccgcttcga gcagacatga taagatacat
     2461 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     2521 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     2581 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt ttttaagcaa
     2641 gtaaaacctc tacaaatgtg gtaaaatctt aattaagact tctaatataa cttcgtatag
     2701 catacattat acgaagttat ggttccactg ttgacaatta atcatcggca tagtatatcg
     2761 gcatagtata atacgacaag gtgaggaact aaacccagga ggcagatcat gattgaacaa
     2821 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     2881 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     2941 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3001 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3061 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3121 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3181 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     3241 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3301 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     3361 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     3421 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     3481 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     3541 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     3601 tgattcgaaa tgaccgacca agcgacgccc aaccggtatc agctcactca aaggcggtaa
     3661 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3721 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3781 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3841 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3901 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3961 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4021 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4081 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4141 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4201 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4261 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4321 agattacgcg cagaaaaaaa ggatttcaag aagatccttt gatcttttct acggggtctg
     4381 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4441 tcttcaccta gatcctttta tagtccggaa tcgtcacaca aaaaaccaac acacagatgt
     4501 aatgaaaata aagatatttt attgcggcca tcgtgatggc tagagtttag ccctcccaca
     4561 cataaccaga gggcagcaat tcacgaatcc caactgccgt cggctgtcca tcactgtcct
     4621 tcactatggc tttgatccca ggatgcagat cgagaagcac ctgtcggcac cgtccgcagg
     4681 ggctcaagat gcccctgttc tcatttccga tcgcgacgat acaagtcagg ttgccagctg
     4741 ccgcagcagc agcagtgccc agcaccacga gttctgcaca aggtccccca gtaaaatgat
     4801 atacattgac accagtgaag atgcggccgt cgctagagag agctgcgctg gcgacgctgt
     4861 agtcttcaga gatggggatg ctgttgattg tagccgttgc tctttcaatg agggtggatt
     4921 cttcttgaga caaaggcttg gccatggtgg cgctagtgtc agaagaatcg agctttttgc
     4981 aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg
     5041 cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg
     5101 aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact
     5161 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     5221 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     5281 actttccaca ccctaactga cacacattcc acagctggtt ctttccgcct cagaaggtac
     5341 ctaaccaagt tcctctttca gaggttattt caggccatgg tgctgcgc
//
