LOCUS       sequence-425908-        7476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human LLGL1. N-terminal HiBiT 
            tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7476)
  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 7476)
  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_LLGL1:1-1064
FEATURES             Location/Qualifiers
     source          1..7476
                     /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             3586..3594
                     /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    4539..4660
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4550..4569)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4775..4808)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4818..4865
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4897..5691
                     /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(4951..4970)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5561..5580
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5849..6437)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5929..5948)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6563..6611
                     /note="synthetic polyadenylation signal"
     CDS             complement(6635..7033)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      7030..7039
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(7064..7421)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7078..7213
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        7230..7421
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7378..7398
                     /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 tgatgaagtt
     1261 tcggttccgg cggcaaggag cagatccgca gcgcgagaaa ctgaaacaag agttgtttgc
     1321 attcaataag accgttgagc acggctttcc caaccagcct agcgcgctcg cgtttgaccc
     1381 ggaattgcgc atcatggcca taggtacccg ttccggcgcg gttaagattt acggtgcccc
     1441 aggcgtagag tttactggac tccatcgcga cgcggcaacc gtgacccaga tgcacttcct
     1501 gactggtcaa ggtcgtttgt tgtcacttct ggacgattcc agtctccatt tgtgggaaat
     1561 agtgcaccat aatggttgcg cccatctcga agaagcgttg tctttccaac tcccgagcag
     1621 gcctggattt gacggtgctt cagccccact cagtcttaca agagtaactg tcgtgcttct
     1681 tgtggcggct tccgacattg ctgcattggg aactgaaggc agttcagtct tctttctcga
     1741 cgtaaccacc ctgaccctgc tggagggcca aaccctcgcg ccaggagagg ttttgcgttc
     1801 ggtaccggat gattatcgtt gtggaaaggc acttggccca gttgaatccc ttcagggaca
     1861 cctgcgtgac ccgacaaaga tacttatcgg ttatagtaga ggcctccttg tcatctggaa
     1921 ccaggcttcg caatgtgttg atcacatatt tctcggcaat caacaacttg aatcgctctg
     1981 ttggggaagg gatagttcca cggtcgtctc gagtcattcg gatggatcct acgccgtatg
     2041 gtcagtagac gccggatctt tcccgacatt gcagcccacg gtcgcgacaa ccccttacgg
     2101 accattcccg tgtaaagcca ttaataagat tttgtggagg aactgcgagt caggcggaca
     2161 cttcatcatt tttagtggcg gtatgccgcg tgcgtcctac ggtgaccgcc actgcgtttc
     2221 agtactgaga gctgaaacac tcgtcacctt ggactttacc agtagaatta ttgacttttt
     2281 cactgtacac tccacaagac ccgaagacga attcgacgat ccccaagcgc tcgccgtact
     2341 gctggaagag gagcttgtgg tactcgatct tcaaacgccc ggttggccgg ctgttccagc
     2401 cccctatctg gcccccctgc atagtagcgc gatcacctgc agtgcccatg ttgcgagcgt
     2461 cccagcgaag ctctgggcca gaattgtcag tgcgggagaa cagcaatctc cccagccggt
     2521 atcttccgct cttagttggc caattacggg cggtcgcaac ctggcgcagg agccgtctca
     2581 acgcggtttg ttgctgaccg gtcatgaaga tggtacagta cgcttctggg atgcatctgg
     2641 tgttgctttg aggcccctct acaaattgtc aaccgccggc ctttttcaga cagattgtga
     2701 acatgcggat tccctggcac aagctgctga ggacgattgg cctcccttca gaaaagtggg
     2761 ctgtttcgac ccctatagtg atgatccgag gcttggtgta caaaaagttg ccttgtgtaa
     2821 atataccgcg cagatggtgg tagcaggcac cgccggtcaa gtgttggtcc ttgagctgag
     2881 cgatgttccc gtggaacagg ctgtttctgt tgcgatcata gaccttttgc aggaccgtga
     2941 aggattcacc tggaagggac atgaacgtct gtcgccgcgc acaggtcctc ttccatggcc
     3001 ggctggtttc caaccccgcg tcctcgtcca atgtcttcct ccagccgccg ttacagcagt
     3061 aacgctccat actgaatggt cactggtggc atttggaaca tctcacggct tcggcctctt
     3121 cgactaccag cgtaagtccc cggtactggc gagatgcacc ttgcatccga atgattccct
     3181 cgcgatggag ggccctctca gcagagtgaa atccctgaag aaatcactcc gtcagtcctt
     3241 tcgtaggata cgcaagagta gagtatctgg aaaaaagcgc gcggccaacg ctagttccaa
     3301 acttcaggaa gccaatgccc aactcgcgga acaagcctgt ccccacgatg tcgaaatgac
     3361 tccagtccag cgcaggattg aaccacgctc agctgacgac tccctttccg gtgttgttag
     3421 gtgcctgtac tttgcggata catttttgcg cgatggcgcc catcacggcc cgaccatgtg
     3481 ggcaggtacc aactctggca gcgttttcgc ttatgcgctt gaagtgcccg ctgccgcagt
     3541 cggaggagaa aagaggccag aacaggcggt cgaggctgtg ttgggtaagg aggtacaact
     3601 catgcaccgc gccccagtag ttgcgattgc ggtcctcgat ggtcgcggcc gtcctctccc
     3661 agaaccgtat gaggcttcaa gggacctcgc tcaggccccg gacatgcaag gcggccacgc
     3721 tgtacttata gcttcggaag aacaattcaa agtgttcact cttccaaagg tgagcgccaa
     3781 gaccaagttc aaattgacgg cacacgaggg ctgcagagtt cgcaaagtag cccttgctac
     3841 tttcgcgtcc gtagcttgcg aagactatgc agaaacctgc ctggcgtgtt tgaccaacct
     3901 tggtgacgtt catgtctttt ctgtgcctgg actgcgccct caggtgcact actcatgtat
     3961 tcgtaaagaa gatatatccg gcattgcctc ctgcgtgttc actcgccatg gccagggctt
     4021 ttatttgata agtcccagcg agttcgagag gttctcgttg tccgcgcgca acataacgga
     4081 gcctctttgt tcgttggata tcaattggcc tagagacgct acccaagcca gttaccgtat
     4141 aagggagtct cccaaactct cacaggccaa tggaacacct tcgattcttt tggctcctca
     4201 gtccctcgac ggtagccctg atccagcaca tagtatggga cctgacacgc cagagccgcc
     4261 tgaggcggcc ctttctccta tgagcataga ttcagcaacg agtgcagata cgactctcga
     4321 tacaaccggc gatgtgacag tagaggatgt caaggacttc cttggtagca gtgaggaatc
     4381 cgagaagaat ctgaggaacc tggctgaaga tgaggcccac gcctgtgcca tcctgatcaa
     4441 ataatctaga ggtcaagctt cgggggttta aacctgcagg catgcaagct gatttggctg
     4501 ctgccaccgc tgagcaaggc cgcttcgagc agacatgata agatacattg atgagtttgg
     4561 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     4621 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     4681 ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta
     4741 caaatgtggt aaaatcttaa ttaagacttc taatataact tcgtatagca tacattatac
     4801 gaagttatgg ttccactgtt gacaattaat catcggcata gtatatcggc atagtataat
     4861 acgacaaggt gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac
     4921 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4981 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     5041 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     5101 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5161 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5221 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5281 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5341 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5401 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5461 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5521 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5581 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5641 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg attcgaaatg
     5701 accgaccaag cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca
     5761 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5821 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5881 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5941 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6001 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6061 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6121 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6181 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6241 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     6301 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6361 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6421 gaaaaaaagg atttcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6481 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6541 tccttttata gtccggaatc gtcacacaaa aaaccaacac acagatgtaa tgaaaataaa
     6601 gatattttat tgcggccatc gtgatggcta gagtttagcc ctcccacaca taaccagagg
     6661 gcagcaattc acgaatccca actgccgtcg gctgtccatc actgtccttc actatggctt
     6721 tgatcccagg atgcagatcg agaagcacct gtcggcaccg tccgcagggg ctcaagatgc
     6781 ccctgttctc atttccgatc gcgacgatac aagtcaggtt gccagctgcc gcagcagcag
     6841 cagtgcccag caccacgagt tctgcacaag gtcccccagt aaaatgatat acattgacac
     6901 cagtgaagat gcggccgtcg ctagagagag ctgcgctggc gacgctgtag tcttcagaga
     6961 tggggatgct gttgattgta gccgttgctc tttcaatgag ggtggattct tcttgagaca
     7021 aaggcttggc catggtggcg ctagtgtcag aagaatcgag ctttttgcaa aagcctaggc
     7081 ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct
     7141 ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag
     7201 ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca
     7261 tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact
     7321 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     7381 ctaactgaca cacattccac agctggttct ttccgcctca gaaggtacct aaccaagttc
     7441 ctctttcaga ggttatttca ggccatggtg ctgcgc
//
