LOCUS       sequence-423416-        7916 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human AHI1. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7916)
  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 7916)
  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_AHI1:1-1196
FEATURES             Location/Qualifiers
     source          1..7916
                     /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    4979..5100
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4990..5009)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(5215..5248)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5258..5305
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5337..6131
                     /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(5391..5410)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6001..6020
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(6289..6877)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6369..6388)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    7003..7051
                     /note="synthetic polyadenylation signal"
     CDS             complement(7075..7473)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      7470..7479
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7504..7861)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7518..7653
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        7670..7861
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7818..7838
                     /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 ccatgcctac agctgagagt
     1261 gaagcaaaag taaaaaccaa agttcgcttt gaagaattgc ttaagaccca cagtgatcta
     1321 atgcgtgaaa agaaaaaact gaagaaaaaa cttgtcaggt ctgaagaaaa catctcacct
     1381 gacactatta gaagcaatct tcactatatg aaagaaacta caagtgatga tcccgacact
     1441 attagaagca atcttcccca tattaaagaa actacaagtg atgatgtaag tgctgctaac
     1501 actaacaacc tgaagaagag cacgagagtc actaaaaaca aattgaggaa cacacagtta
     1561 gcaactgaaa atcctaatgg tgatgctagt gtagaggaag acaaacaagg aaagccaaat
     1621 aaaaaggtga taaagacggt gccccagttg actacacaag acctgaaacc ggaaactcct
     1681 gagaataagg ttgattctac acaccagaaa acacatacaa agccacagcc aggcgttgat
     1741 catcagaaaa gtgagaaggc aaatgaggga agagaagaga ctgatttaga agaggatgaa
     1801 gaattgatgc aagcatatca gtgccatgta actgaagaaa tggcaaagga gattaagagg
     1861 aaaataagaa agaaactgaa agaacagttg acttactttc cctcagatac tttattccat
     1921 gatgacaaac taagcagtga aaaaaggaaa aagaaaaagg aagttccagt cttctctaaa
     1981 gctgaaacaa gtacattgac catctctggt gacacagttg aaggtgaaca aaagaaagaa
     2041 tcttcagtta gatcagtttc ttcagattct catcaagatg atgaaataag ctcaatggaa
     2101 caaagcacag aagacagcat gcaagatgat acaaaaccta aaccaaaaaa aacaaaaaag
     2161 aagactaaag cagttgcaga taataatgaa gatgttgatg gtgatggtgt tcatgaaata
     2221 acaagccgag atagcccggt ttatcccaaa tgtttgcttg atgatgacct tgtcttggga
     2281 gtttacattc accgaactga tagacttaag tcagatttta tgatttctca cccaatggta
     2341 aaaattcatg tggttgatga gcatactggt caatatgtca agaaagatga tagtggacgg
     2401 cctgtttcat cttactatga aaaagagaat gtggattata ttcttcctat tatgacccag
     2461 ccatatgatt ttaaacagtt aaaatcaaga cttccagagt gggaagaaca aattgtattt
     2521 aatgaaaatt ttccctattt gcttcgaggc tctgatgaga gtcctaaagt catcctgttc
     2581 tttgagattc ttgatttctt aagcgtggat gaaattaaga ataattctga ggttcaaaac
     2641 caagaatgtg gctttcggaa aattgcctgg gcatttctta agcttctggg agccaatgga
     2701 aatgcaaaca tcaactcaaa acttcgcttg cagctatatt acccacctac taagcctcga
     2761 tccccattaa gtgttgttga ggcatttgaa tggtggtcaa aatgtccaag aaatcattac
     2821 ccatcaacac tgtacgtaac tgtaagagga ctgaaagttc cagactgtat aaagccatct
     2881 taccgctcta tgatggctct tcaggaggaa aaaggtaaac cagtgcattg tgaacgtcac
     2941 catgagtcaa gctcagtaga cacagaacct ggattagaag agtcaaagga agtaataaag
     3001 tggaaacgac tccctgggca ggcttgccgt atcccaaaca aacacctctt ctcactaaat
     3061 gcaggagaac gaggatgttt ttgtcttgat ttctcccaca atggaagaat attagcagca
     3121 gcttgtgcca gccgggatgg atatccaatt attttatatg aaattccttc tggacgtttc
     3181 atgagagaat tgtgtggcca cctcaatatc atttatgatc tttcctggtc aaaagatgat
     3241 cactacatcc ttacttcatc atctgatggc actgccagga tatggaaaaa tgaaataaac
     3301 aatacaaata ctttcagagt tttacctcat ccttcttttg tttacacggc taaattccat
     3361 ccagctgtaa gagagctagt agttacagga tgctatgatt ccatgatacg gatatggaaa
     3421 gttgagatga gagaagattc tgccatattg gtccgacagt ttgatgttca caaaagtttt
     3481 atcaactcac tttgttttga tactgaaggt catcatatgt attcaggaga ttgtacaggg
     3541 gtgattgttg tttggaatac ctatgtcaag attaatgatt tggaacattc agtgcaccac
     3601 tggactataa ataaggaaat taaagaaact gagtttaagg gaattccaat aagttatttg
     3661 gagattcatc ccaatggaaa acgtttgtta atccatacca aagacagtac tttgagaatt
     3721 atggatctcc ggatattagt agcaaggaag tttgtaggag cagcaaatta tcgggagaag
     3781 attcatagta ctttgactcc atgtgggact tttctgtttg ctggaagtga ggatggtata
     3841 gtgtatgttt ggaacccaga aacaggagaa caagtagcca tgtattctga cttgccattc
     3901 aagtcaccca ttcgagacat ttcttatcat ccatttgaaa atatggttgc attctgtgca
     3961 tttgggcaaa atgagccaat tcttctgtat atttacgatt tccatgttgc ccagcaggag
     4021 gctgaaatgt tcaaacgcta caatggaaca tttccattac ctggaataca ccaaagtcaa
     4081 gatgccctat gtacctgtcc aaaactaccc catcaaggct cttttcagat tgatgaattt
     4141 gtccacactg aaagttcttc aacgaagatg cagctagtaa aacagaggct tgaaactgtc
     4201 acagaggtga tacgttcctg tgctgcaaaa gtcaacaaaa atctctcatt tacttcacca
     4261 ccagcagttt cctcacaaca gtctaagtta aagcagtcaa acatgctgac cgctcaagag
     4321 attctacatc agtttggttt cactcagacc gggattatca gcatagaaag aaagccttgt
     4381 aaccatcagg tagatacagc accaacggta gtggctcttt atgactacac agcgaatcga
     4441 tcagatgaac taaccatcca tcgcggagac attatccgag tgtttttcaa agataatgaa
     4501 gactggtggt atggcagcat aggaaaggga caggaaggtt attttccagc taatcatgtg
     4561 gctagtgaaa cactgtatca agaactgcct cctgagataa aggagcgatc ccctccttta
     4621 agccctgagg aaaaaactaa aatagaaaaa tctccagctc ctcaaaagca atcaatcaat
     4681 aagaacaagt cccaggactt cagactaggc tcagaatcta tgacacattc tgaaatgaga
     4741 aaagaacaga gccatgagga ccaaggacac ataatggata cacggatgag gaagaacaag
     4801 caagcaggca gaaaagtcac tctaatagag ggctcgagcg gtgggagctc cggtgtgagc
     4861 ggctggcggc tgttcaagaa gattagctaa gtttaaacgg ccgcgactct agactgcagg
     4921 catgcaagct gatttggctg ctgccaccgc tgagcaaggc cgcttcgagc agacatgata
     4981 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     5041 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     5101 aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt
     5161 ttaagcaagt aaaacctcta caaatgtggt aaaatcttaa ttaagacttc taatataact
     5221 tcgtatagca tacattatac gaagttatgg ttccactgtt gacaattaat catcggcata
     5281 gtatatcggc atagtataat acgacaaggt gaggaactaa acccaggagg cagatcatga
     5341 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     5401 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     5461 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     5521 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     5581 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     5641 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     5701 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5761 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5821 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     5881 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5941 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     6001 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     6061 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     6121 agttcttctg attcgaaatg accgaccaag cgacgcccaa ccggtatcag ctcactcaaa
     6181 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6241 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6301 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6361 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6421 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6481 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6541 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6601 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6661 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6721 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6781 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6841 caagcagcag attacgcgca gaaaaaaagg atttcaagaa gatcctttga tcttttctac
     6901 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6961 aaaaaggatc ttcacctaga tccttttata gtccggaatc gtcacacaaa aaaccaacac
     7021 acagatgtaa tgaaaataaa gatattttat tgcggccatc gtgatggcta gagtttagcc
     7081 ctcccacaca taaccagagg gcagcaattc acgaatccca actgccgtcg gctgtccatc
     7141 actgtccttc actatggctt tgatcccagg atgcagatcg agaagcacct gtcggcaccg
     7201 tccgcagggg ctcaagatgc ccctgttctc atttccgatc gcgacgatac aagtcaggtt
     7261 gccagctgcc gcagcagcag cagtgcccag caccacgagt tctgcacaag gtcccccagt
     7321 aaaatgatat acattgacac cagtgaagat gcggccgtcg ctagagagag ctgcgctggc
     7381 gacgctgtag tcttcagaga tggggatgct gttgattgta gccgttgctc tttcaatgag
     7441 ggtggattct tcttgagaca aaggcttggc catggtggcg ctagtgtcag aagaatcgag
     7501 ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga
     7561 ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga
     7621 atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt
     7681 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     7741 tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     7801 gcctggggac tttccacacc ctaactgaca cacattccac agctggttct ttccgcctca
     7861 gaaggtacct aaccaagttc ctctttcaga ggttatttca ggccatggtg ctgcgc
//
