LOCUS       sequence-423414-        5966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human AAAS. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5966)
  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 5966)
  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_AAAS:1-546
FEATURES             Location/Qualifiers
     source          1..5966
                     /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    3029..3150
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3040..3059)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3265..3298)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3308..3355
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3387..4181
                     /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(3441..3460)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4051..4070
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(4339..4927)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4419..4438)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5053..5101
                     /note="synthetic polyadenylation signal"
     CDS             complement(5125..5523)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      5520..5529
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5554..5911)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5568..5703
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5720..5911
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5868..5888
                     /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 ccatgtgctc tctggggttg
     1261 ttccctcctc caccgcctcg gggtcaagtc accctatatg agcacaataa cgagctggtg
     1321 acgggcagta gctatgagag cccgcccccc gacttccggg gccagtggat caatcttcct
     1381 gtcctacaac tgacaaagga tcccctaaag acccctggaa ggctggacca tggcacaaga
     1441 actgccttca tccatcaccg ggagcaagtg tggaagagat gcatcaacat ttggcgtgat
     1501 gtgggccttt ttggggtgct aaatgaaatt gcaaactcag aagaagaggt gtttgagtgg
     1561 gtgaagacgg catccggctg ggccctggca ctctgtcgat gggcctcttc cctccatggg
     1621 tccctgttcc cccatctgtc tctcaggagc gaagatctga tcgctgaatt tgcccaagtc
     1681 acaaattggt ccagctgctg cttgcgtgtc tttgcatggc acccccacac caacaagttt
     1741 gcagtggccc tgctagatga ctcagtccgt gtgtataatg ccagcagcac catagtcccc
     1801 tccctgaagc accggctgca gcgaaatgtg gcgtctctgg cctggaagcc ccttagtgcc
     1861 tctgtcttgg ctgtggcctg ccagagctgc attcttatct ggaccctgga ccctacctcc
     1921 ttgtctaccc gaccctcttc tggctgtgcc caagtgctgt ctcaccctgg gcatacacct
     1981 gttaccagct tggcctgggc ccccagtggg gggcggctgc tctcagcttc acccgtggat
     2041 gctgctatcc gggtatggga tgtctcaaca gagacctgtg tcccccttcc ctggtttcga
     2101 ggaggtgggg tgaccaacct gctctggtcc ccagacggca gcaaaatcct ggctaccact
     2161 ccttcagctg tctttcgagt ctgggaggcc cagatgtgga cttgtgagag gtggcctact
     2221 ctatcagggc gctgtcagac tggctgctgg agcccagatg gcagccgact gctgttcact
     2281 gtattgggag agccactgat ttactccctg tcttttccag aacgttgtgg tgagggaaag
     2341 gggtgcgttg gaggtgcaaa gtcagcaacg attgtggcag atctgtctga gacaacaata
     2401 cagacaccag atggtgagga gaggcttggg ggagaggctc actccatggt ctgggacccc
     2461 agtggggaac gtctggctgt gcttatgaaa ggaaagccaa gggtacagga tggtaaacca
     2521 gtcatcctcc tttttcgcac tcgaaacagc cctgtgtttg agctccttcc ctgtggcatt
     2581 atccaggggg agccaggagc ccagccccag ctcatcactt tccatccttc cttcaacaaa
     2641 ggggccctgc tcagtgtggg ctggtccaca ggccgaattg cccacatccc gctgtacttt
     2701 gtcaatgccc agtttccacg ttttagccca gtgcttgggc gggcccagga accccctgct
     2761 gggggtggag gctctattca tgacctgccc ctctttactg agacatcccc aacctctgcc
     2821 ccttgggacc ctctcccagg gccaccacct gttctgcccc actccccaca ttcccacctc
     2881 ggctcgagcg gtgggagctc cggtgtgagc ggctggcggc tgttcaagaa gattagctaa
     2941 gtttaaacgg ccgcgactct agactgcagg catgcaagct gatttggctg ctgccaccgc
     3001 tgagcaaggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     3061 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     3121 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     3181 caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta caaatgtggt
     3241 aaaatcttaa ttaagacttc taatataact tcgtatagca tacattatac gaagttatgg
     3301 ttccactgtt gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt
     3361 gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac gcaggttctc
     3421 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     3481 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     3541 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     3601 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     3661 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3721 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3781 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3841 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3901 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3961 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4021 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4081 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4141 agcgcatcgc cttctatcgc cttcttgacg agttcttctg attcgaaatg accgaccaag
     4201 cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4261 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4321 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4381 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4441 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4501 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4561 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4621 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4681 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4741 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     4801 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4861 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4921 atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4981 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttata
     5041 gtccggaatc gtcacacaaa aaaccaacac acagatgtaa tgaaaataaa gatattttat
     5101 tgcggccatc gtgatggcta gagtttagcc ctcccacaca taaccagagg gcagcaattc
     5161 acgaatccca actgccgtcg gctgtccatc actgtccttc actatggctt tgatcccagg
     5221 atgcagatcg agaagcacct gtcggcaccg tccgcagggg ctcaagatgc ccctgttctc
     5281 atttccgatc gcgacgatac aagtcaggtt gccagctgcc gcagcagcag cagtgcccag
     5341 caccacgagt tctgcacaag gtcccccagt aaaatgatat acattgacac cagtgaagat
     5401 gcggccgtcg ctagagagag ctgcgctggc gacgctgtag tcttcagaga tggggatgct
     5461 gttgattgta gccgttgctc tttcaatgag ggtggattct tcttgagaca aaggcttggc
     5521 catggtggcg ctagtgtcag aagaatcgag ctttttgcaa aagcctaggc ctccaaaaaa
     5581 gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat
     5641 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
     5701 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
     5761 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg
     5821 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
     5881 cacattccac agctggttct ttccgcctca gaaggtacct aaccaagttc ctctttcaga
     5941 ggttatttca ggccatggtg ctgcgc
//
