LOCUS       sequence-423437-        6908 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human MED16 isoform 3. 
            C-terminal HiBiT tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6908)
  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 6908)
  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_MED16:1-860
FEATURES             Location/Qualifiers
     source          1..6908
                     /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    3971..4092
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3982..4001)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(4207..4240)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4250..4297
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4329..5123
                     /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(4383..4402)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4993..5012
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(5281..5869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5361..5380)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5995..6043
                     /note="synthetic polyadenylation signal"
     CDS             complement(6067..6465)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     regulatory      6462..6471
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(6496..6853)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6510..6645
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        6662..6853
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     6810..6830
                     /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 ccatgtgtga tttgcggcgg
     1261 ccagcggcag gtgggatgat ggacttggcc tacgtctgtg agtgggagaa atggtccaag
     1321 agcacccact gcccatcggt gcccctggcc tgcgcctggt cctgccgaaa tctcatcgcc
     1381 ttcaccatgg acctgcgcag cgatgaccag gacctgaccc gcatgatcca catcctggac
     1441 acggagcacc cctgggacct gcactcgatc ccctcagagc accacgaggc catcacctgc
     1501 ctggagtggg accagtcagg ctcccggctc ctgtcagcag atgccgacgg gcagatcaag
     1561 tgctggagca tggcggacca cctggctaat agctgggaga gctcagtggg cagcctagtg
     1621 gagggggacc ccattgtggc cctgtcctgg ctgcacaatg gtgtgaaact ggccctgcac
     1681 gtggagaagt cgggcgcctc cagcttcggg gagaagttct cccgagtcaa gttctcaccg
     1741 tcgctcacgc tgttcggcgg caagcccatg gagggctgga tcgcggtgac ggtcagcggc
     1801 ctggtcaccg tgtccctgct gaagcccagc gggcaggtgc tgacgtccac cgagagcctg
     1861 tgccggctgc gcggccgcgt ggccctggcc gacatcgcct tcaccggcgg cggcaacatc
     1921 gtggtggcca cggcggacgg cagcagcgcg tcgcccgtgc agttctacaa ggtgtgcgtg
     1981 agcgtggtga gcgagaagtg ccgtatcgac acggagatcc tgccctccct gttcatgcgc
     2041 tgcaccaccg acctcaaccg caaggacaag tttcccgcca tcacccacct caagttcctg
     2101 gcccgggaca tgtcggagca ggtgcttttg tgcgcgtcca gccagaccag cagcatcgtg
     2161 gagtgctggt ccctgcgcaa ggagggactc cccgtgaaca acatcttcca gcagatctcc
     2221 cccgtggttg gcgacaaaca gcccacaatt ctcaaatggc ggatcctatc ggccaccaac
     2281 gatctggacc gtgtgtcggc cgtggcgctg cccaagctgc ccatctcgct caccaacacc
     2341 gacctcaagg tggccagcga cacacagttc taccctggcc tcgggctggc cctggccttc
     2401 cacgacggca gcgtccacat cgtgcaccgg ctctcactgc agaccatggc cgtcttctac
     2461 agctccgcgg ccccgaggcc tgtggatgag ccggccatga agcgcccccg caccgcgggc
     2521 cccgccgtcc acttaaaggc tatgcagcta tcgtggacgt cactggccct ggtggggatt
     2581 gacagccacg ggaagctgag cgtgctccgc ctctcacctt ccatgggcca cccgctggag
     2641 gtggggctgg cgctgcggca cctgctcttc ctgctggagt actgcatggt gaccggctac
     2701 gactggtggg acatcctgct gcacgtgcag cccagtatgg tacagagcct ggtggagaag
     2761 ctgcacgagg agtacacgcg ccagaccgct gccctgcagc aggtcctctc cacccggatc
     2821 ctggccatga aggcctcgct ctgcaagctg tcgccctgca cggtgacccg cgtgtgcgac
     2881 taccacacca agctcttcct catcgccatc agctccaccc tgaagtcgct gctgcgcccc
     2941 cactttctca acacgcctga caagagcccc ggcgaccggc tgaccgagat ctgcaccaag
     3001 atcaccgacg tcgacattga caaggtcatg atcaacctca agacggagga atttgtgctg
     3061 gacatgaaca cactgcaggc gctgcagcag ctcttgcagt gggtgggcga cttcgtgctg
     3121 tacctgctgg ccagcctacc caaccagggt tccctgctga ggccgggcca cagctttctg
     3181 cgggacggca cctcgctggg catgcttcgg gaattgatgg tggtcatccg catctggggc
     3241 cttctgaagc ccagctgcct gcccgtgtat acggccacct cggataccca ggacagcatg
     3301 tccctgctct tccgcctgct caccaagctc tggatctgct ttcctagcac aggtccatgc
     3361 tctgtgtggg tgcttttggg atggcagcca cttccaggtc gcgatgaggg cccagcgagc
     3421 gagccggacg aggcgctggt ggatgaatgc tgcctgctgc ccagccagct gcttatcccc
     3481 agcctggact ggctgccagc cagcgacggc ctggttagcc gcctgcagcc caagcagccc
     3541 cttcgtctgc agtttggccg ggcgcccacg ctgcctggca gtgctgccac cctgcagctc
     3601 gacggcctcg ccagggcccc aggccagccc aagatcgacc acctgcggag gctgcacctt
     3661 ggcgcttgcc ccacggagga atgcaaggcc tgcaccaggt gcggctgtgt caccatgctc
     3721 aagtcgccca acagaaccac ggcggtgaag cagtgggagc agcgctggat caagaactgc
     3781 ctgtgcggtg ggctctggtg gcgggtgccc ctcagctacc ccggctcgag cggtgggagc
     3841 tccggtgtga gcggctggcg gctgttcaag aagattagct aagtttaaac ggccgcgact
     3901 ctagactgca ggcatgcaag ctgatttggc tgctgccacc gctgagcaag gccgcttcga
     3961 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     4021 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     4081 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg
     4141 tgggaggttt ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatctt aattaagact
     4201 tctaatataa cttcgtatag catacattat acgaagttat ggttccactg ttgacaatta
     4261 atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact aaacccagga
     4321 ggcagatcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4381 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4441 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4501 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4561 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4621 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4681 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4741 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4801 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     4861 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4921 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4981 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5041 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5101 gccttcttga cgagttcttc tgattcgaaa tgaccgacca agcgacgccc aaccggtatc
     5161 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5221 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5281 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5341 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5401 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5461 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5521 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5581 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5641 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5701 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     5761 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5821 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt
     5881 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5941 catgagatta tcaaaaagga tcttcaccta gatcctttta tagtccggaa tcgtcacaca
     6001 aaaaaccaac acacagatgt aatgaaaata aagatatttt attgcggcca tcgtgatggc
     6061 tagagtttag ccctcccaca cataaccaga gggcagcaat tcacgaatcc caactgccgt
     6121 cggctgtcca tcactgtcct tcactatggc tttgatccca ggatgcagat cgagaagcac
     6181 ctgtcggcac cgtccgcagg ggctcaagat gcccctgttc tcatttccga tcgcgacgat
     6241 acaagtcagg ttgccagctg ccgcagcagc agcagtgccc agcaccacga gttctgcaca
     6301 aggtccccca gtaaaatgat atacattgac accagtgaag atgcggccgt cgctagagag
     6361 agctgcgctg gcgacgctgt agtcttcaga gatggggatg ctgttgattg tagccgttgc
     6421 tctttcaatg agggtggatt cttcttgaga caaaggcttg gccatggtgg cgctagtgtc
     6481 agaagaatcg agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg
     6541 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
     6601 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg
     6661 cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     6721 gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct
     6781 gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc acagctggtt
     6841 ctttccgcct cagaaggtac ctaaccaagt tcctctttca gaggttattt caggccatgg
     6901 tgctgcgc
//
