LOCUS       sequence-425909-        6864 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human MED16 isoform 3. 
            N-terminal HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6864)
  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 6864)
  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_MED16:1-860
FEATURES             Location/Qualifiers
     source          1..6864
                     /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    3927..4048
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3938..3957)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4163..4196)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4206..4253
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4285..5079
                     /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(4339..4358)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4949..4968
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5237..5825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5317..5336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5951..5999
                     /note="synthetic polyadenylation signal"
     CDS             complement(6023..6421)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6418..6427
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6452..6809)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6466..6601
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6618..6809
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6766..6786
                     /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 tgtgtgattt
     1261 gcggcggcca gcggcaggtg ggatgatgga cttggcctac gtctgtgagt gggagaaatg
     1321 gtccaagagc acccactgcc catcggtgcc cctggcctgc gcctggtcct gccgaaatct
     1381 catcgccttc accatggacc tgcgcagcga tgaccaggac ctgacccgca tgatccacat
     1441 cctggacacg gagcacccct gggacctgca ctcgatcccc tcagagcacc acgaggccat
     1501 cacctgcctg gagtgggacc agtcaggctc ccggctcctg tcagcagatg ccgacgggca
     1561 gatcaagtgc tggagcatgg cggaccacct ggctaatagc tgggagagct cagtgggcag
     1621 cctagtggag ggggacccca ttgtggccct gtcctggctg cacaatggtg tgaaactggc
     1681 cctgcacgtg gagaagtcgg gcgcctccag cttcggggag aagttctccc gagtcaagtt
     1741 ctcaccgtcg ctcacgctgt tcggcggcaa gcccatggag ggctggatcg cggtgacggt
     1801 cagcggcctg gtcaccgtgt ccctgctgaa gcccagcggg caggtgctga cgtccaccga
     1861 gagcctgtgc cggctgcgcg gccgcgtggc cctggccgac atcgccttca ccggcggcgg
     1921 caacatcgtg gtggccacgg cggacggcag cagcgcgtcg cccgtgcagt tctacaaggt
     1981 gtgcgtgagc gtggtgagcg agaagtgccg tatcgacacg gagatcctgc cctccctgtt
     2041 catgcgctgc accaccgacc tcaaccgcaa ggacaagttt cccgccatca cccacctcaa
     2101 gttcctggcc cgggacatgt cggagcaggt gcttttgtgc gcgtccagcc agaccagcag
     2161 catcgtggag tgctggtccc tgcgcaagga gggactcccc gtgaacaaca tcttccagca
     2221 gatctccccc gtggttggcg acaaacagcc cacaattctc aaatggcgga tcctatcggc
     2281 caccaacgat ctggaccgtg tgtcggccgt ggcgctgccc aagctgccca tctcgctcac
     2341 caacaccgac ctcaaggtgg ccagcgacac acagttctac cctggcctcg ggctggccct
     2401 ggccttccac gacggcagcg tccacatcgt gcaccggctc tcactgcaga ccatggccgt
     2461 cttctacagc tccgcggccc cgaggcctgt ggatgagccg gccatgaagc gcccccgcac
     2521 cgcgggcccc gccgtccact taaaggctat gcagctatcg tggacgtcac tggccctggt
     2581 ggggattgac agccacggga agctgagcgt gctccgcctc tcaccttcca tgggccaccc
     2641 gctggaggtg gggctggcgc tgcggcacct gctcttcctg ctggagtact gcatggtgac
     2701 cggctacgac tggtgggaca tcctgctgca cgtgcagccc agtatggtac agagcctggt
     2761 ggagaagctg cacgaggagt acacgcgcca gaccgctgcc ctgcagcagg tcctctccac
     2821 ccggatcctg gccatgaagg cctcgctctg caagctgtcg ccctgcacgg tgacccgcgt
     2881 gtgcgactac cacaccaagc tcttcctcat cgccatcagc tccaccctga agtcgctgct
     2941 gcgcccccac tttctcaaca cgcctgacaa gagccccggc gaccggctga ccgagatctg
     3001 caccaagatc accgacgtcg acattgacaa ggtcatgatc aacctcaaga cggaggaatt
     3061 tgtgctggac atgaacacac tgcaggcgct gcagcagctc ttgcagtggg tgggcgactt
     3121 cgtgctgtac ctgctggcca gcctacccaa ccagggttcc ctgctgaggc cgggccacag
     3181 ctttctgcgg gacggcacct cgctgggcat gcttcgggaa ttgatggtgg tcatccgcat
     3241 ctggggcctt ctgaagccca gctgcctgcc cgtgtatacg gccacctcgg atacccagga
     3301 cagcatgtcc ctgctcttcc gcctgctcac caagctctgg atctgctttc ctagcacagg
     3361 tccatgctct gtgtgggtgc ttttgggatg gcagccactt ccaggtcgcg atgagggccc
     3421 agcgagcgag ccggacgagg cgctggtgga tgaatgctgc ctgctgccca gccagctgct
     3481 tatccccagc ctggactggc tgccagccag cgacggcctg gttagccgcc tgcagcccaa
     3541 gcagcccctt cgtctgcagt ttggccgggc gcccacgctg cctggcagtg ctgccaccct
     3601 gcagctcgac ggcctcgcca gggccccagg ccagcccaag atcgaccacc tgcggaggct
     3661 gcaccttggc gcttgcccca cggaggaatg caaggcctgc accaggtgcg gctgtgtcac
     3721 catgctcaag tcgcccaaca gaaccacggc ggtgaagcag tgggagcagc gctggatcaa
     3781 gaactgcctg tgcggtgggc tctggtggcg ggtgcccctc agctacccct aatctagagg
     3841 tcaagcttcg ggggtttaaa cctgcaggca tgcaagctga tttggctgct gccaccgctg
     3901 agcaaggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     3961 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     4021 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     4081 ggttcagggg gagatgtggg aggttttttt aagcaagtaa aacctctaca aatgtggtaa
     4141 aatcttaatt aagacttcta atataacttc gtatagcata cattatacga agttatggtt
     4201 ccactgttga caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga
     4261 ggaactaaac ccaggaggca gatcatgatt gaacaagatg gattgcacgc aggttctccg
     4321 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     4381 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     4441 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     4501 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     4561 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     4621 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4681 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4741 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4801 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4861 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4921 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4981 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5041 cgcatcgcct tctatcgcct tcttgacgag ttcttctgat tcgaaatgac cgaccaagcg
     5101 acgcccaacc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     5161 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     5221 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     5281 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5341 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     5401 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     5461 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     5521 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     5581 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5641 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     5701 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5761 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5821 ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5881 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttatagt
     5941 ccggaatcgt cacacaaaaa accaacacac agatgtaatg aaaataaaga tattttattg
     6001 cggccatcgt gatggctaga gtttagccct cccacacata accagagggc agcaattcac
     6061 gaatcccaac tgccgtcggc tgtccatcac tgtccttcac tatggctttg atcccaggat
     6121 gcagatcgag aagcacctgt cggcaccgtc cgcaggggct caagatgccc ctgttctcat
     6181 ttccgatcgc gacgatacaa gtcaggttgc cagctgccgc agcagcagca gtgcccagca
     6241 ccacgagttc tgcacaaggt cccccagtaa aatgatatac attgacacca gtgaagatgc
     6301 ggccgtcgct agagagagct gcgctggcga cgctgtagtc ttcagagatg gggatgctgt
     6361 tgattgtagc cgttgctctt tcaatgaggg tggattcttc ttgagacaaa ggcttggcca
     6421 tggtggcgct agtgtcagaa gaatcgagct ttttgcaaaa gcctaggcct ccaaaaaagc
     6481 ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa
     6541 aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga
     6601 tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac
     6661 ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca
     6721 tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca
     6781 cattccacag ctggttcttt ccgcctcaga aggtacctaa ccaagttcct ctttcagagg
     6841 ttatttcagg ccatggtgct gcgc
//
