LOCUS       sequence-425954-        7736 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human PAF1. C-terminal HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7736)
  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 7736)
  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_PAF1:1-531
FEATURES             Location/Qualifiers
     source          1..7736
                     /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             2690..2710
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2726..3610
                     /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(3682..3700)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3737..3858
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3748..3767)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3802..3821
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4103..4460
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4103..4294
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4126..4146)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4311..4446
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4511..5305
                     /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(4565..4584)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5175..5194
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5369..5417
                     /note="synthetic polyadenylation signal"
     promoter        5573..5677
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5678..5746,5747..6538)
                     /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(5896..5915)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6696..7284)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6776..6795)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7400..7683
                     /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 catggcgccc accatccaga cccaggccca gcgggaggat ggccacaggc ccaattccca
     1141 ccggactctg cctgagaggt ctggagtggt ctgccgagtc aagtactgca atagcctccc
     1201 tgatatcccc ttcgacccca agttcatcac ctaccccttc gaccagaaca ggttcgtcca
     1261 gtacaaagcc acttccttgg agaaacagca caaacatgac ctcctgactg agccagacct
     1321 gggggtcacc atcgatctca tcaatcctga cacctaccgc atcgacccca atgttcttct
     1381 agatccagct gatgagaaac ttttggaaga ggagattcag gcccccacca gctccaagag
     1441 atcccagcag cacgcgaagg tggtgccatg gatgcgaaag acagagtaca tctccactga
     1501 gttcaaccgt tatggcatct ccaatgagaa gcctgaggtc aagattgggg tttctgtgaa
     1561 gcagcagttt accgaggaag aaatatacaa agacagggat agccagatca cagccattga
     1621 gaagactttt gaggatgccc agaaatcaat ctcacagcat tacagcaaac cccgagtcac
     1681 accggtggag gtcatgcctg tcttcccaga ctttaagatg tggatcaatc catgtgctca
     1741 ggtgatcttt gactcagacc cagcccccaa ggacacgagt ggtgcagctg cgttggagat
     1801 gatgtctcag gccatgatta ggggcatgat ggatgaggaa gggaaccagt ttgtggccta
     1861 tttcctgcct gtagaagaga cgttgaagaa acgaaagcgg gaccaggagg aggagatgga
     1921 ctatgcacca gatgatgtgt atgactacaa aattgctcgg gagtacaact ggaacgtgaa
     1981 gaacaaagct agcaagggct atgaggaaaa ctacttcttc atcttccgag agggtgacgg
     2041 ggtttactac aatgagttgg aaaccagggt ccgccttagt aagcgccggg ccaaggctgg
     2101 ggttcagtca ggcaccaacg ccctgcttgt ggtcaaacat cgggacatga atgagaagga
     2161 actggaagct caggaggcac ggaaggccca gctagaaaac cacgaaccgg aggaggaaga
     2221 ggaagaggag atggagacag aagagaaaga agctgggggc tcagatgagg agcaggagaa
     2281 gggcagcagc agtgagaagg agggcagtga agatgagcac tcgggcagcg agagtgaacg
     2341 ggaggaaggt gacagggacg aggccagtga caagagtggc agtggtgagg acgagagcag
     2401 cgaggatgag gcccgggctg cccgtgacaa agaggagatc tttggcagtg atgctgattc
     2461 tgaggacgat gccgactctg atgatgagga cagaggacag gcccaaggtg gcagtgacaa
     2521 tgattcagac agcggcagca atgggggtgg ccagcggagc cggagccaca gccgcagcgc
     2581 cagtcccttc cccagtggca gcgagcactc ggcccaggag gatggcagtg aagctgcagc
     2641 ttctgattcc agtgaagctg atagtgacag tgacctcgag ccaaccactg aggatctgta
     2701 ctttcagagc gataacgatg gatccgaaat cggtactggc tttccattcg acccccatta
     2761 tgtggaagtc ctgggcgagc gcatgcacta cgtcgatgtt ggtccgcgcg atggcacccc
     2821 tgtgctgttc ctgcacggta acccgacctc ctcctacgtg tggcgcaaca tcatcccgca
     2881 tgttgcaccg acccatcgct gcattgctcc agacctgatc ggtatgggca aatccgacaa
     2941 accagacctg ggttatttct tcgacgacca cgtccgcttc atggatgcct tcatcgaagc
     3001 cctgggtctg gaagaggtcg tcctggtcat tcacgactgg ggctccgctc tgggtttcca
     3061 ctgggccaag cgcaatccag agcgcgtcaa aggtattgca tttatggagt tcatccgccc
     3121 tatcccgacc tgggacgaat ggccagaatt tgcccgcgag accttccagg ccttccgcac
     3181 caccgacgtc ggccgcaagc tgatcatcga tcagaacgtt tttatcgagg gtacgctgcc
     3241 gatgggtgtc gtccgcccgc tgactgaagt cgagatggac cattaccgcg agccgttcct
     3301 gaatcctgtt gaccgcgagc cactgtggcg cttcccaaac gagctgccaa tcgccggtga
     3361 gccagcgaac atcgtcgcgc tggtcgaaga atacatggac tggctgcacc agtcccctgt
     3421 cccgaagctg ctgttctggg gcaccccagg cgttctgatc ccaccggccg aagccgctcg
     3481 cctggccaaa agcctgccta actgcaaggc tgtggacatc ggcccgggtc tgaatctgct
     3541 gcaagaagac aacccggacc tgatcggcag cgagatcgcg cgctggctgt ctactctgga
     3601 gatttccggt taatagaatt ggcatgcaag ctgatccggc tgctaacaaa gcccgaaagg
     3661 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcggccg
     3721 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     3781 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     3841 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     3901 gagatgtggg aggttttttt aagcaagtaa aacctctaca aatgtggtaa aatcgaattt
     3961 taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc
     4021 ggtatttcac accgcatacg cggatctgcg cagcaccatg gcctgaaata acctctgaaa
     4081 gaggaacttg gttaggtacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     4141 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4201 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4261 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4321 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4381 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4441 gaggcctagg cttttgcaaa aagcttgatt cttctgacac aacagtctcg aacttaaggc
     4501 tagagccacc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4561 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4621 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4681 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4741 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4801 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4861 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4921 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4981 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     5041 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5101 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5161 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5221 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5281 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     5341 acgcccaacc tgccatcacg atggccgcaa taaaatatct ttattttcat tacatctgtg
     5401 tgttggtttt ttgtgtgaat cgatagcgat aaggatcctc tttgcgcttg cgttttccct
     5461 tgtccagata gcccagtagc tgacattcat ccggggtcag caccgtttct gcggactggc
     5521 tttctacgta atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     5581 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5641 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5701 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5761 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5821 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5881 cactttcaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5941 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     6001 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     6061 tgataacact gcggccaact tacttctgac aactatcgga ggaccgaagg agctaaccgc
     6121 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     6181 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6241 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6301 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6361 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6421 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6481 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaatt
     6541 cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg
     6601 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     6661 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6721 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6781 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6841 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6901 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6961 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     7021 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     7081 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     7141 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     7201 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     7261 acgcgcagaa aaaaaggatt tcaagaagat cctttgatct tttctacggg gtctgacgct
     7321 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     7381 acctagatcc ttttatagtc cggaaataca ggaacgcacg ctggatggcc cttcgctggg
     7441 atggtgaaac catgaaaaat ggcagcttca gtggattaag tgggggtaat gtggcctgta
     7501 ccctctggtt gcataggtat tcatacggtt aaaatttatc aggcgcgatt gcggcagttt
     7561 ttcgggtggt ttgttgccat ttttacctgt ctgctgccgt gatcgcgctg aacgcgtttt
     7621 agcggtgcgt acaattaagg gattatggta aatccactta ctgtctgccc tcgtagccat
     7681 cgagataaac cgcagtactc cggccacgat gcgtccggcg tagaggatcg agatct
//
