LOCUS       sequence-423436-        7520 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human LLGL1. C-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7520)
  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 7520)
  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_LLGL1:1-1064
FEATURES             Location/Qualifiers
     source          1..7520
                     /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"
     RBS             3579..3587
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    4583..4704
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4594..4613)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4819..4852)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4862..4909
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4941..5735
                     /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(4995..5014)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5605..5624
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5893..6481)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5973..5992)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6607..6655
                     /note="synthetic polyadenylation signal"
     CDS             complement(6679..7077)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      7074..7083
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7108..7465)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7122..7257
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        7274..7465
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7422..7442
                     /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 ccatgatgaa gtttcggttc
     1261 cggcggcaag gagcagatcc gcagcgcgag aaactgaaac aagagttgtt tgcattcaat
     1321 aagaccgttg agcacggctt tcccaaccag cctagcgcgc tcgcgtttga cccggaattg
     1381 cgcatcatgg ccataggtac ccgttccggc gcggttaaga tttacggtgc cccaggcgta
     1441 gagtttactg gactccatcg cgacgcggca accgtgaccc agatgcactt cctgactggt
     1501 caaggtcgtt tgttgtcact tctggacgat tccagtctcc atttgtggga aatagtgcac
     1561 cataatggtt gcgcccatct cgaagaagcg ttgtctttcc aactcccgag caggcctgga
     1621 tttgacggtg cttcagcccc actcagtctt acaagagtaa ctgtcgtgct tcttgtggcg
     1681 gcttccgaca ttgctgcatt gggaactgaa ggcagttcag tcttctttct cgacgtaacc
     1741 accctgaccc tgctggaggg ccaaaccctc gcgccaggag aggttttgcg ttcggtaccg
     1801 gatgattatc gttgtggaaa ggcacttggc ccagttgaat cccttcaggg acacctgcgt
     1861 gacccgacaa agatacttat cggttatagt agaggcctcc ttgtcatctg gaaccaggct
     1921 tcgcaatgtg ttgatcacat atttctcggc aatcaacaac ttgaatcgct ctgttgggga
     1981 agggatagtt ccacggtcgt ctcgagtcat tcggatggat cctacgccgt atggtcagta
     2041 gacgccggat ctttcccgac attgcagccc acggtcgcga caacccctta cggaccattc
     2101 ccgtgtaaag ccattaataa gattttgtgg aggaactgcg agtcaggcgg acacttcatc
     2161 atttttagtg gcggtatgcc gcgtgcgtcc tacggtgacc gccactgcgt ttcagtactg
     2221 agagctgaaa cactcgtcac cttggacttt accagtagaa ttattgactt tttcactgta
     2281 cactccacaa gacccgaaga cgaattcgac gatccccaag cgctcgccgt actgctggaa
     2341 gaggagcttg tggtactcga tcttcaaacg cccggttggc cggctgttcc agccccctat
     2401 ctggcccccc tgcatagtag cgcgatcacc tgcagtgccc atgttgcgag cgtcccagcg
     2461 aagctctggg ccagaattgt cagtgcggga gaacagcaat ctccccagcc ggtatcttcc
     2521 gctcttagtt ggccaattac gggcggtcgc aacctggcgc aggagccgtc tcaacgcggt
     2581 ttgttgctga ccggtcatga agatggtaca gtacgcttct gggatgcatc tggtgttgct
     2641 ttgaggcccc tctacaaatt gtcaaccgcc ggcctttttc agacagattg tgaacatgcg
     2701 gattccctgg cacaagctgc tgaggacgat tggcctccct tcagaaaagt gggctgtttc
     2761 gacccctata gtgatgatcc gaggcttggt gtacaaaaag ttgccttgtg taaatatacc
     2821 gcgcagatgg tggtagcagg caccgccggt caagtgttgg tccttgagct gagcgatgtt
     2881 cccgtggaac aggctgtttc tgttgcgatc atagaccttt tgcaggaccg tgaaggattc
     2941 acctggaagg gacatgaacg tctgtcgccg cgcacaggtc ctcttccatg gccggctggt
     3001 ttccaacccc gcgtcctcgt ccaatgtctt cctccagccg ccgttacagc agtaacgctc
     3061 catactgaat ggtcactggt ggcatttgga acatctcacg gcttcggcct cttcgactac
     3121 cagcgtaagt ccccggtact ggcgagatgc accttgcatc cgaatgattc cctcgcgatg
     3181 gagggccctc tcagcagagt gaaatccctg aagaaatcac tccgtcagtc ctttcgtagg
     3241 atacgcaaga gtagagtatc tggaaaaaag cgcgcggcca acgctagttc caaacttcag
     3301 gaagccaatg cccaactcgc ggaacaagcc tgtccccacg atgtcgaaat gactccagtc
     3361 cagcgcagga ttgaaccacg ctcagctgac gactcccttt ccggtgttgt taggtgcctg
     3421 tactttgcgg atacattttt gcgcgatggc gcccatcacg gcccgaccat gtgggcaggt
     3481 accaactctg gcagcgtttt cgcttatgcg cttgaagtgc ccgctgccgc agtcggagga
     3541 gaaaagaggc cagaacaggc ggtcgaggct gtgttgggta aggaggtaca actcatgcac
     3601 cgcgccccag tagttgcgat tgcggtcctc gatggtcgcg gccgtcctct cccagaaccg
     3661 tatgaggctt caagggacct cgctcaggcc ccggacatgc aaggcggcca cgctgtactt
     3721 atagcttcgg aagaacaatt caaagtgttc actcttccaa aggtgagcgc caagaccaag
     3781 ttcaaattga cggcacacga gggctgcaga gttcgcaaag tagcccttgc tactttcgcg
     3841 tccgtagctt gcgaagacta tgcagaaacc tgcctggcgt gtttgaccaa ccttggtgac
     3901 gttcatgtct tttctgtgcc tggactgcgc cctcaggtgc actactcatg tattcgtaaa
     3961 gaagatatat ccggcattgc ctcctgcgtg ttcactcgcc atggccaggg cttttatttg
     4021 ataagtccca gcgagttcga gaggttctcg ttgtccgcgc gcaacataac ggagcctctt
     4081 tgttcgttgg atatcaattg gcctagagac gctacccaag ccagttaccg tataagggag
     4141 tctcccaaac tctcacaggc caatggaaca ccttcgattc ttttggctcc tcagtccctc
     4201 gacggtagcc ctgatccagc acatagtatg ggacctgaca cgccagagcc gcctgaggcg
     4261 gccctttctc ctatgagcat agattcagca acgagtgcag atacgactct cgatacaacc
     4321 ggcgatgtga cagtagagga tgtcaaggac ttccttggta gcagtgagga atccgagaag
     4381 aatctgagga acctggctga agatgaggcc cacgcctgtg ccatcctgat caaaggctcg
     4441 agcggtggga gctccggtgt gagcggctgg cggctgttca agaagattag ctaagtttaa
     4501 acggccgcga ctctagactg caggcatgca agctgatttg gctgctgcca ccgctgagca
     4561 aggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
     4621 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     4681 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     4741 cagggggaga tgtgggaggt ttttttaagc aagtaaaacc tctacaaatg tggtaaaatc
     4801 ttaattaaga cttctaatat aacttcgtat agcatacatt atacgaagtt atggttccac
     4861 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
     4921 ctaaacccag gaggcagatc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4981 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5041 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5101 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     5161 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5221 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5281 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5341 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5401 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5461 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5521 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5581 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5641 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5701 tcgccttcta tcgccttctt gacgagttct tctgattcga aatgaccgac caagcgacgc
     5761 ccaaccggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5821 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5881 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5941 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     6001 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     6061 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     6121 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6181 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6241 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6301 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     6361 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6421 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatttca
     6481 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6541 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tatagtccgg
     6601 aatcgtcaca caaaaaacca acacacagat gtaatgaaaa taaagatatt ttattgcggc
     6661 catcgtgatg gctagagttt agccctccca cacataacca gagggcagca attcacgaat
     6721 cccaactgcc gtcggctgtc catcactgtc cttcactatg gctttgatcc caggatgcag
     6781 atcgagaagc acctgtcggc accgtccgca ggggctcaag atgcccctgt tctcatttcc
     6841 gatcgcgacg atacaagtca ggttgccagc tgccgcagca gcagcagtgc ccagcaccac
     6901 gagttctgca caaggtcccc cagtaaaatg atatacattg acaccagtga agatgcggcc
     6961 gtcgctagag agagctgcgc tggcgacgct gtagtcttca gagatgggga tgctgttgat
     7021 tgtagccgtt gctctttcaa tgagggtgga ttcttcttga gacaaaggct tggccatggt
     7081 ggcgctagtg tcagaagaat cgagcttttt gcaaaagcct aggcctccaa aaaagcctcc
     7141 tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa
     7201 aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg
     7261 cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct
     7321 gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct
     7381 ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt
     7441 ccacagctgg ttctttccgc ctcagaaggt acctaaccaa gttcctcttt cagaggttat
     7501 ttcaggccat ggtgctgcgc
//
