LOCUS       sequence-425936-        5304 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human GNB1. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5304)
  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 5304)
  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_GNB1:1-340
FEATURES             Location/Qualifiers
     source          1..5304
                     /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    2367..2488
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2378..2397)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2603..2636)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2646..2693
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2725..3519
                     /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(2779..2798)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3389..3408
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(3677..4265)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3757..3776)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4391..4439
                     /note="synthetic polyadenylation signal"
     CDS             complement(4463..4861)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      4858..4867
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4892..5249)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4906..5041
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        5058..5249
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5206..5226
                     /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 tgagtgagct
     1261 tgaccagtta cggcaggagg ccgagcaact taagaaccag attcgagacg ccaggaaagc
     1321 atgtgcagat gcaactctct ctcagatcac aaacaacatc gacccagtgg gaagaatcca
     1381 aatgcgcacg aggaggacac tgcgggggca cctggccaag atctacgcca tgcactgggg
     1441 cacagactcc aggcttctcg tcagtgcctc gcaggatggt aaacttatca tctgggacag
     1501 ctacaccacc aacaaggtcc acgccatccc tctgcgctcc tcctgggtca tgacctgtgc
     1561 atatgcccct tctgggaact atgtggcctg cggtggcctg gataacattt gctccattta
     1621 caatctgaaa actcgtgagg ggaacgtgcg cgtgagtcgt gagctggcag gacacacagg
     1681 ttacctgtcc tgctgccgat tcctggatga caatcagatc gtcaccagct ctggagacac
     1741 cacgtgtgcc ctgtgggaca tcgagaccgg ccagcagacg accacgttta ccggacacac
     1801 tggagatgtc atgagccttt ctcttgctcc tgacaccaga ctgttcgtct ctggtgcttg
     1861 tgatgcttca gccaaactct gggatgtgcg agaaggcatg tgccggcaga ccttcactgg
     1921 ccacgagtct gacatcaatg ccatttgctt ctttccaaat ggcaatgcat ttgccactgg
     1981 ctcagacgac gccacctgca ggctgtttga ccttcgtgct gaccaggagc tcatgactta
     2041 ctcccatgac aacatcatct gcgggatcac ctctgtctcc ttctccaaga gcgggcgcct
     2101 cctccttgct gggtacgacg acttcaactg caacgtctgg gatgcactca aagccgaccg
     2161 ggcaggtgtc ttggctgggc atgacaaccg cgtcagctgc ctgggcgtga ctgacgatgg
     2221 catggctgtg gcgacagggt cctgggatag cttcctcaag atctggaact aatctagagg
     2281 tcaagcttcg ggggtttaaa cctgcaggca tgcaagctga tttggctgct gccaccgctg
     2341 agcaaggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     2401 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     2461 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     2521 ggttcagggg gagatgtggg aggttttttt aagcaagtaa aacctctaca aatgtggtaa
     2581 aatcttaatt aagacttcta atataacttc gtatagcata cattatacga agttatggtt
     2641 ccactgttga caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga
     2701 ggaactaaac ccaggaggca gatcatgatt gaacaagatg gattgcacgc aggttctccg
     2761 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     2821 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     2881 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     2941 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3001 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3061 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3121 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3181 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3241 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3301 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3361 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3421 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3481 cgcatcgcct tctatcgcct tcttgacgag ttcttctgat tcgaaatgac cgaccaagcg
     3541 acgcccaacc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3601 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3661 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3721 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3781 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3841 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3901 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3961 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4021 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4081 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     4141 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4201 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     4261 ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     4321 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttatagt
     4381 ccggaatcgt cacacaaaaa accaacacac agatgtaatg aaaataaaga tattttattg
     4441 cggccatcgt gatggctaga gtttagccct cccacacata accagagggc agcaattcac
     4501 gaatcccaac tgccgtcggc tgtccatcac tgtccttcac tatggctttg atcccaggat
     4561 gcagatcgag aagcacctgt cggcaccgtc cgcaggggct caagatgccc ctgttctcat
     4621 ttccgatcgc gacgatacaa gtcaggttgc cagctgccgc agcagcagca gtgcccagca
     4681 ccacgagttc tgcacaaggt cccccagtaa aatgatatac attgacacca gtgaagatgc
     4741 ggccgtcgct agagagagct gcgctggcga cgctgtagtc ttcagagatg gggatgctgt
     4801 tgattgtagc cgttgctctt tcaatgaggg tggattcttc ttgagacaaa ggcttggcca
     4861 tggtggcgct agtgtcagaa gaatcgagct ttttgcaaaa gcctaggcct ccaaaaaagc
     4921 ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa
     4981 aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga
     5041 tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac
     5101 ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca
     5161 tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca
     5221 cattccacag ctggttcttt ccgcctcaga aggtacctaa ccaagttcct ctttcagagg
     5281 ttatttcagg ccatggtgct gcgc
//
