LOCUS       sequence-425892-        7872 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human AHI1. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7872)
  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 7872)
  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_AHI1:1-1196
FEATURES             Location/Qualifiers
     source          1..7872
                     /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    4935..5056
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4946..4965)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(5171..5204)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5214..5261
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5293..6087
                     /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(5347..5366)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5957..5976
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(6245..6833)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6325..6344)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6959..7007
                     /note="synthetic polyadenylation signal"
     CDS             complement(7031..7429)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      7426..7435
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7460..7817)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7474..7609
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        7626..7817
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7774..7794
                     /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 tgcctacagc
     1261 tgagagtgaa gcaaaagtaa aaaccaaagt tcgctttgaa gaattgctta agacccacag
     1321 tgatctaatg cgtgaaaaga aaaaactgaa gaaaaaactt gtcaggtctg aagaaaacat
     1381 ctcacctgac actattagaa gcaatcttca ctatatgaaa gaaactacaa gtgatgatcc
     1441 cgacactatt agaagcaatc ttccccatat taaagaaact acaagtgatg atgtaagtgc
     1501 tgctaacact aacaacctga agaagagcac gagagtcact aaaaacaaat tgaggaacac
     1561 acagttagca actgaaaatc ctaatggtga tgctagtgta gaggaagaca aacaaggaaa
     1621 gccaaataaa aaggtgataa agacggtgcc ccagttgact acacaagacc tgaaaccgga
     1681 aactcctgag aataaggttg attctacaca ccagaaaaca catacaaagc cacagccagg
     1741 cgttgatcat cagaaaagtg agaaggcaaa tgagggaaga gaagagactg atttagaaga
     1801 ggatgaagaa ttgatgcaag catatcagtg ccatgtaact gaagaaatgg caaaggagat
     1861 taagaggaaa ataagaaaga aactgaaaga acagttgact tactttccct cagatacttt
     1921 attccatgat gacaaactaa gcagtgaaaa aaggaaaaag aaaaaggaag ttccagtctt
     1981 ctctaaagct gaaacaagta cattgaccat ctctggtgac acagttgaag gtgaacaaaa
     2041 gaaagaatct tcagttagat cagtttcttc agattctcat caagatgatg aaataagctc
     2101 aatggaacaa agcacagaag acagcatgca agatgataca aaacctaaac caaaaaaaac
     2161 aaaaaagaag actaaagcag ttgcagataa taatgaagat gttgatggtg atggtgttca
     2221 tgaaataaca agccgagata gcccggttta tcccaaatgt ttgcttgatg atgaccttgt
     2281 cttgggagtt tacattcacc gaactgatag acttaagtca gattttatga tttctcaccc
     2341 aatggtaaaa attcatgtgg ttgatgagca tactggtcaa tatgtcaaga aagatgatag
     2401 tggacggcct gtttcatctt actatgaaaa agagaatgtg gattatattc ttcctattat
     2461 gacccagcca tatgatttta aacagttaaa atcaagactt ccagagtggg aagaacaaat
     2521 tgtatttaat gaaaattttc cctatttgct tcgaggctct gatgagagtc ctaaagtcat
     2581 cctgttcttt gagattcttg atttcttaag cgtggatgaa attaagaata attctgaggt
     2641 tcaaaaccaa gaatgtggct ttcggaaaat tgcctgggca tttcttaagc ttctgggagc
     2701 caatggaaat gcaaacatca actcaaaact tcgcttgcag ctatattacc cacctactaa
     2761 gcctcgatcc ccattaagtg ttgttgaggc atttgaatgg tggtcaaaat gtccaagaaa
     2821 tcattaccca tcaacactgt acgtaactgt aagaggactg aaagttccag actgtataaa
     2881 gccatcttac cgctctatga tggctcttca ggaggaaaaa ggtaaaccag tgcattgtga
     2941 acgtcaccat gagtcaagct cagtagacac agaacctgga ttagaagagt caaaggaagt
     3001 aataaagtgg aaacgactcc ctgggcaggc ttgccgtatc ccaaacaaac acctcttctc
     3061 actaaatgca ggagaacgag gatgtttttg tcttgatttc tcccacaatg gaagaatatt
     3121 agcagcagct tgtgccagcc gggatggata tccaattatt ttatatgaaa ttccttctgg
     3181 acgtttcatg agagaattgt gtggccacct caatatcatt tatgatcttt cctggtcaaa
     3241 agatgatcac tacatcctta cttcatcatc tgatggcact gccaggatat ggaaaaatga
     3301 aataaacaat acaaatactt tcagagtttt acctcatcct tcttttgttt acacggctaa
     3361 attccatcca gctgtaagag agctagtagt tacaggatgc tatgattcca tgatacggat
     3421 atggaaagtt gagatgagag aagattctgc catattggtc cgacagtttg atgttcacaa
     3481 aagttttatc aactcacttt gttttgatac tgaaggtcat catatgtatt caggagattg
     3541 tacaggggtg attgttgttt ggaataccta tgtcaagatt aatgatttgg aacattcagt
     3601 gcaccactgg actataaata aggaaattaa agaaactgag tttaagggaa ttccaataag
     3661 ttatttggag attcatccca atggaaaacg tttgttaatc cataccaaag acagtacttt
     3721 gagaattatg gatctccgga tattagtagc aaggaagttt gtaggagcag caaattatcg
     3781 ggagaagatt catagtactt tgactccatg tgggactttt ctgtttgctg gaagtgagga
     3841 tggtatagtg tatgtttgga acccagaaac aggagaacaa gtagccatgt attctgactt
     3901 gccattcaag tcacccattc gagacatttc ttatcatcca tttgaaaata tggttgcatt
     3961 ctgtgcattt gggcaaaatg agccaattct tctgtatatt tacgatttcc atgttgccca
     4021 gcaggaggct gaaatgttca aacgctacaa tggaacattt ccattacctg gaatacacca
     4081 aagtcaagat gccctatgta cctgtccaaa actaccccat caaggctctt ttcagattga
     4141 tgaatttgtc cacactgaaa gttcttcaac gaagatgcag ctagtaaaac agaggcttga
     4201 aactgtcaca gaggtgatac gttcctgtgc tgcaaaagtc aacaaaaatc tctcatttac
     4261 ttcaccacca gcagtttcct cacaacagtc taagttaaag cagtcaaaca tgctgaccgc
     4321 tcaagagatt ctacatcagt ttggtttcac tcagaccggg attatcagca tagaaagaaa
     4381 gccttgtaac catcaggtag atacagcacc aacggtagtg gctctttatg actacacagc
     4441 gaatcgatca gatgaactaa ccatccatcg cggagacatt atccgagtgt ttttcaaaga
     4501 taatgaagac tggtggtatg gcagcatagg aaagggacag gaaggttatt ttccagctaa
     4561 tcatgtggct agtgaaacac tgtatcaaga actgcctcct gagataaagg agcgatcccc
     4621 tcctttaagc cctgaggaaa aaactaaaat agaaaaatct ccagctcctc aaaagcaatc
     4681 aatcaataag aacaagtccc aggacttcag actaggctca gaatctatga cacattctga
     4741 aatgagaaaa gaacagagcc atgaggacca aggacacata atggatacac ggatgaggaa
     4801 gaacaagcaa gcaggcagaa aagtcactct aatagagtaa tctagaggtc aagcttcggg
     4861 ggtttaaacc tgcaggcatg caagctgatt tggctgctgc caccgctgag caaggccgct
     4921 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     4981 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     5041 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     5101 gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcttaattaa
     5161 gacttctaat ataacttcgt atagcataca ttatacgaag ttatggttcc actgttgaca
     5221 attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg aactaaaccc
     5281 aggaggcaga tcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     5341 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     5401 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     5461 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     5521 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     5581 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     5641 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     5701 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     5761 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     5821 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     5881 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     5941 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     6001 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     6061 tatcgccttc ttgacgagtt cttctgattc gaaatgaccg accaagcgac gcccaaccgg
     6121 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6181 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6241 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6301 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6361 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6421 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6481 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6541 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6601 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6661 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     6721 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6781 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggattt caagaagatc
     6841 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6901 tggtcatgag attatcaaaa aggatcttca cctagatcct tttatagtcc ggaatcgtca
     6961 cacaaaaaac caacacacag atgtaatgaa aataaagata ttttattgcg gccatcgtga
     7021 tggctagagt ttagccctcc cacacataac cagagggcag caattcacga atcccaactg
     7081 ccgtcggctg tccatcactg tccttcacta tggctttgat cccaggatgc agatcgagaa
     7141 gcacctgtcg gcaccgtccg caggggctca agatgcccct gttctcattt ccgatcgcga
     7201 cgatacaagt caggttgcca gctgccgcag cagcagcagt gcccagcacc acgagttctg
     7261 cacaaggtcc cccagtaaaa tgatatacat tgacaccagt gaagatgcgg ccgtcgctag
     7321 agagagctgc gctggcgacg ctgtagtctt cagagatggg gatgctgttg attgtagccg
     7381 ttgctctttc aatgagggtg gattcttctt gagacaaagg cttggccatg gtggcgctag
     7441 tgtcagaaga atcgagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact
     7501 tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc
     7561 agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta
     7621 ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg
     7681 gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg ctttgcatac
     7741 ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca ttccacagct
     7801 ggttctttcc gcctcagaag gtacctaacc aagttcctct ttcagaggtt atttcaggcc
     7861 atggtgctgc gc
//
