LOCUS       sequence-425890-        5922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human AAAS. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5922)
  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 5922)
  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_AAAS:1-546
FEATURES             Location/Qualifiers
     source          1..5922
                     /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    2985..3106
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2996..3015)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3221..3254)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3264..3311
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3343..4137
                     /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(3397..3416)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4007..4026
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4295..4883)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4375..4394)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5009..5057
                     /note="synthetic polyadenylation signal"
     CDS             complement(5081..5479)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5476..5485
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5510..5867)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5524..5659
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5676..5867
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5824..5844
                     /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 tgtgctctct
     1261 ggggttgttc cctcctccac cgcctcgggg tcaagtcacc ctatatgagc acaataacga
     1321 gctggtgacg ggcagtagct atgagagccc gccccccgac ttccggggcc agtggatcaa
     1381 tcttcctgtc ctacaactga caaaggatcc cctaaagacc cctggaaggc tggaccatgg
     1441 cacaagaact gccttcatcc atcaccggga gcaagtgtgg aagagatgca tcaacatttg
     1501 gcgtgatgtg ggcctttttg gggtgctaaa tgaaattgca aactcagaag aagaggtgtt
     1561 tgagtgggtg aagacggcat ccggctgggc cctggcactc tgtcgatggg cctcttccct
     1621 ccatgggtcc ctgttccccc atctgtctct caggagcgaa gatctgatcg ctgaatttgc
     1681 ccaagtcaca aattggtcca gctgctgctt gcgtgtcttt gcatggcacc cccacaccaa
     1741 caagtttgca gtggccctgc tagatgactc agtccgtgtg tataatgcca gcagcaccat
     1801 agtcccctcc ctgaagcacc ggctgcagcg aaatgtggcg tctctggcct ggaagcccct
     1861 tagtgcctct gtcttggctg tggcctgcca gagctgcatt cttatctgga ccctggaccc
     1921 tacctccttg tctacccgac cctcttctgg ctgtgcccaa gtgctgtctc accctgggca
     1981 tacacctgtt accagcttgg cctgggcccc cagtgggggg cggctgctct cagcttcacc
     2041 cgtggatgct gctatccggg tatgggatgt ctcaacagag acctgtgtcc cccttccctg
     2101 gtttcgagga ggtggggtga ccaacctgct ctggtcccca gacggcagca aaatcctggc
     2161 taccactcct tcagctgtct ttcgagtctg ggaggcccag atgtggactt gtgagaggtg
     2221 gcctactcta tcagggcgct gtcagactgg ctgctggagc ccagatggca gccgactgct
     2281 gttcactgta ttgggagagc cactgattta ctccctgtct tttccagaac gttgtggtga
     2341 gggaaagggg tgcgttggag gtgcaaagtc agcaacgatt gtggcagatc tgtctgagac
     2401 aacaatacag acaccagatg gtgaggagag gcttggggga gaggctcact ccatggtctg
     2461 ggaccccagt ggggaacgtc tggctgtgct tatgaaagga aagccaaggg tacaggatgg
     2521 taaaccagtc atcctccttt ttcgcactcg aaacagccct gtgtttgagc tccttccctg
     2581 tggcattatc cagggggagc caggagccca gccccagctc atcactttcc atccttcctt
     2641 caacaaaggg gccctgctca gtgtgggctg gtccacaggc cgaattgccc acatcccgct
     2701 gtactttgtc aatgcccagt ttccacgttt tagcccagtg cttgggcggg cccaggaacc
     2761 ccctgctggg ggtggaggct ctattcatga cctgcccctc tttactgaga catccccaac
     2821 ctctgcccct tgggaccctc tcccagggcc accacctgtt ctgccccact ccccacattc
     2881 ccacctctaa tctagaggtc aagcttcggg ggtttaaacc tgcaggcatg caagctgatt
     2941 tggctgctgc caccgctgag caaggccgct tcgagcagac atgataagat acattgatga
     3001 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3061 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3121 cattcatttt atgtttcagg ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa
     3181 cctctacaaa tgtggtaaaa tcttaattaa gacttctaat ataacttcgt atagcataca
     3241 ttatacgaag ttatggttcc actgttgaca attaatcatc ggcatagtat atcggcatag
     3301 tataatacga caaggtgagg aactaaaccc aggaggcaga tcatgattga acaagatgga
     3361 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3421 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     3481 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     3541 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     3601 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     3661 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     3721 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     3781 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     3841 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     3901 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     3961 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4021 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4081 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgattc
     4141 gaaatgaccg accaagcgac gcccaaccgg tatcagctca ctcaaaggcg gtaatacggt
     4201 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4261 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4321 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4381 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4441 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4501 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4561 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4621 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4681 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     4741 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4801 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4861 cgcgcagaaa aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4921 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4981 cctagatcct tttatagtcc ggaatcgtca cacaaaaaac caacacacag atgtaatgaa
     5041 aataaagata ttttattgcg gccatcgtga tggctagagt ttagccctcc cacacataac
     5101 cagagggcag caattcacga atcccaactg ccgtcggctg tccatcactg tccttcacta
     5161 tggctttgat cccaggatgc agatcgagaa gcacctgtcg gcaccgtccg caggggctca
     5221 agatgcccct gttctcattt ccgatcgcga cgatacaagt caggttgcca gctgccgcag
     5281 cagcagcagt gcccagcacc acgagttctg cacaaggtcc cccagtaaaa tgatatacat
     5341 tgacaccagt gaagatgcgg ccgtcgctag agagagctgc gctggcgacg ctgtagtctt
     5401 cagagatggg gatgctgttg attgtagccg ttgctctttc aatgagggtg gattcttctt
     5461 gagacaaagg cttggccatg gtggcgctag tgtcagaaga atcgagcttt ttgcaaaagc
     5521 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct
     5581 cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg
     5641 gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga
     5701 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg
     5761 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     5821 cacaccctaa ctgacacaca ttccacagct ggttctttcc gcctcagaag gtacctaacc
     5881 aagttcctct ttcagaggtt atttcaggcc atggtgctgc gc
//
