LOCUS       sequence-423326-        7586 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human C9orf72. C-terminal 
            HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7586)
  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 7586)
  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: pHTC-HaloTag_C9orf72:1-481
FEATURES             Location/Qualifiers
     source          1..7586
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2540..2560
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2576..3460
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     primer_bind     complement(3532..3550)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3587..3708
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3598..3617)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3652..3671
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3953..4310
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3953..4144
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3976..3996)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4161..4296
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4361..5155
                     /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(4415..4434)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5025..5044
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5219..5267
                     /note="synthetic polyadenylation signal"
     promoter        5423..5527
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5528..5596,5597..6388)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5746..5765)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6546..7134)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6626..6645)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7250..7533
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcaaa gcgatcgctt ccgaattcac
     1081 catgtcgact ctttgcccac cgccatctcc agctgttgcc aagacagaga ttgctttaag
     1141 tggcaaatca cctttattag cagctacttt tgcttactgg gacaatattc ttggtcctag
     1201 agtaaggcac atttgggctc caaagacaga acaggtactt ctcagtgatg gagaaataac
     1261 ttttcttgcc aaccacactc taaatggaga aatccttcga aatgcagaga gtggtgctat
     1321 agatgtaaag ttttttgtct tgtctgaaaa gggagtgatt attgtttcat taatctttga
     1381 tggaaactgg aatggggatc gcagcacata tggactatca attatacttc cacagacaga
     1441 acttagtttc tacctcccac ttcatagagt gtgtgttgat agattaacac atataatccg
     1501 gaaaggaaga atatggatgc ataaggaaag acaagaaaat gtccagaaga ttatcttaga
     1561 aggcacagag agaatggaag atcagggtca gagtattatt ccaatgctta ctggagaagt
     1621 gattcctgta atggaactgc tttcatctat gaaatcacac agtgttcctg aagaaataga
     1681 tatagctgat acagtactca atgatgatga tattggtgac agctgtcatg aaggctttct
     1741 tctcaatgcc atcagctcac acttgcaaac ctgtggctgt tccgttgtag taggtagcag
     1801 tgcagagaaa gtaaataaga tagtcagaac attatgcctt tttctgactc cagcagagag
     1861 aaaatgctcc aggttatgtg aagcagaatc atcatttaaa tatgagtcag ggctctttgt
     1921 acaaggcctg ctaaaggatt caactggaag ctttgtgctg cctttccggc aagtcatgta
     1981 tgctccatat cccaccacac acatagatgt ggatgtcaat actgtgaagc agatgccacc
     2041 ctgtcatgaa catatttata atcagcgtag atacatgaga tccgagctga cagccttctg
     2101 gagagccact tcagaagaag acatggctca ggatacgatc atctacactg acgaaagctt
     2161 tactcctgat ttgaatattt ttcaagatgt cttacacaga gacactctag tgaaagcctt
     2221 cctggatcag gtctttcagc tgaaacctgg cttatctctc agaagtactt tccttgcaca
     2281 gtttctactt gtccttcaca gaaaagcctt gacactaata aaatatatag aagacgatac
     2341 gcagaaggga aaaaagccct ttaaatctct tcggaacctg aagatagacc ttgatttaac
     2401 agcagagggc gatcttaaca taataatggc tctggctgag aaaattaaac caggcctaca
     2461 ctcttttatc tttggaagac ctttctacac tagtgtgcaa gaacgagatg ttctaatgac
     2521 ttttctcgag ccaaccactg aggatctgta ctttcagagc gataacgatg gatccgaaat
     2581 cggtactggc tttccattcg acccccatta tgtggaagtc ctgggcgagc gcatgcacta
     2641 cgtcgatgtt ggtccgcgcg atggcacccc tgtgctgttc ctgcacggta acccgacctc
     2701 ctcctacgtg tggcgcaaca tcatcccgca tgttgcaccg acccatcgct gcattgctcc
     2761 agacctgatc ggtatgggca aatccgacaa accagacctg ggttatttct tcgacgacca
     2821 cgtccgcttc atggatgcct tcatcgaagc cctgggtctg gaagaggtcg tcctggtcat
     2881 tcacgactgg ggctccgctc tgggtttcca ctgggccaag cgcaatccag agcgcgtcaa
     2941 aggtattgca tttatggagt tcatccgccc tatcccgacc tgggacgaat ggccagaatt
     3001 tgcccgcgag accttccagg ccttccgcac caccgacgtc ggccgcaagc tgatcatcga
     3061 tcagaacgtt tttatcgagg gtacgctgcc gatgggtgtc gtccgcccgc tgactgaagt
     3121 cgagatggac cattaccgcg agccgttcct gaatcctgtt gaccgcgagc cactgtggcg
     3181 cttcccaaac gagctgccaa tcgccggtga gccagcgaac atcgtcgcgc tggtcgaaga
     3241 atacatggac tggctgcacc agtcccctgt cccgaagctg ctgttctggg gcaccccagg
     3301 cgttctgatc ccaccggccg aagccgctcg cctggccaaa agcctgccta actgcaaggc
     3361 tgtggacatc ggcccgggtc tgaatctgct gcaagaagac aacccggacc tgatcggcag
     3421 cgagatcgcg cgctggctgt ctactctgga gatttccggt taatagaatt ggcatgcaag
     3481 ctgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
     3541 aataactagc ataacccctt ggggcggccg cttcgagcag acatgataag atacattgat
     3601 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     3661 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     3721 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggttttttt aagcaagtaa
     3781 aacctctaca aatgtggtaa aatcgaattt taacaaaata ttaacgctta caatttcctg
     3841 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg cggatctgcg
     3901 cagcaccatg gcctgaaata acctctgaaa gaggaacttg gttaggtacc ttctgaggcg
     3961 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4021 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4081 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4141 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4201 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     4261 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttgatt
     4321 cttctgacac aacagtctcg aacttaaggc tagagccacc atgattgaac aagatggatt
     4381 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     4441 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     4501 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     4561 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4621 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4681 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4741 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4801 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4861 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     4921 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4981 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     5041 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     5101 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     5161 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg atggccgcaa
     5221 taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagcgat
     5281 aaggatcctc tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat
     5341 ccggggtcag caccgtttct gcggactggc tttctacgta atggtttctt agacgtcagg
     5401 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5461 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5521 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5581 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5641 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5701 tcgccccgaa gaacgttttc caatgatgag cactttcaaa gttctgctat gtggcgcggt
     5761 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5821 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5881 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5941 aactatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     6001 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     6061 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     6121 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     6181 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6241 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6301 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6361 aggtgcctca ctgattaagc attggtaatt cgaaatgacc gaccaagcga cgcccaaccg
     6421 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6481 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6541 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6601 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6661 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6721 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6781 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6841 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6901 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6961 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     7021 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     7081 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat
     7141 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     7201 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttatagtc cggaaataca
     7261 ggaacgcacg ctggatggcc cttcgctggg atggtgaaac catgaaaaat ggcagcttca
     7321 gtggattaag tgggggtaat gtggcctgta ccctctggtt gcataggtat tcatacggtt
     7381 aaaatttatc aggcgcgatt gcggcagttt ttcgggtggt ttgttgccat ttttacctgt
     7441 ctgctgccgt gatcgcgctg aacgcgtttt agcggtgcgt acaattaagg gattatggta
     7501 aatccactta ctgtctgccc tcgtagccat cgagataaac cgcagtactc cggccacgat
     7561 gcgtccggcg tagaggatcg agatct
//
