LOCUS       sequence-423309-        7268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human ACTB. C-terminal HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7268)
  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 7268)
  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_ACTB:1-375
FEATURES             Location/Qualifiers
     source          1..7268
                     /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             2222..2242
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2258..3142
                     /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(3214..3232)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3269..3390
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3280..3299)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3334..3353
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3635..3992
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3635..3826
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3658..3678)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3843..3978
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4043..4837
                     /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(4097..4116)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4707..4726
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4901..4949
                     /note="synthetic polyadenylation signal"
     promoter        5105..5209
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5210..5278,5279..6070)
                     /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(5428..5447)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6228..6816)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6308..6327)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6932..7215
                     /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 catggatgat gatatcgccg cgctcgtcgt cgacaacggc tccggcatgt gcaaggccgg
     1141 cttcgcgggc gacgatgccc cccgggccgt cttcccctcc atcgtggggc gccccaggca
     1201 ccagggcgtg atggtgggca tgggtcagaa ggattcctat gtgggcgacg aggcccagag
     1261 caagagaggc atcctcaccc tgaagtaccc catcgagcac ggcatcgtca ccaactggga
     1321 cgacatggag aaaatctggc accacacctt ctacaatgag ctgcgtgtgg ctcccgagga
     1381 gcaccccgtg ctgctgaccg aggcccccct gaaccccaag gccaaccgcg agaagatgac
     1441 ccagatcatg tttgagacct tcaacacccc agccatgtac gttgctatcc aggctgtgct
     1501 atccctgtac gcctctggcc gtaccactgg catcgtgatg gactccggtg acggggtcac
     1561 ccacactgtg cccatctacg aggggtatgc cctcccccat gccatcctgc gtctggacct
     1621 ggctggccgg gacctgactg actacctcat gaagatcctc accgagcgcg gctacagctt
     1681 caccaccacg gccgagcggg aaatcgtgcg tgacattaag gagaagctgt gctacgtcgc
     1741 cctggacttc gagcaagaga tggccacggc tgcttccagc tcctccctgg agaagagcta
     1801 cgagctgcct gacggccagg tcatcaccat tggcaatgag cggttccgct gccctgaggc
     1861 actcttccag ccttccttcc tgggcatgga gtcctgtggc atccacgaaa ctaccttcaa
     1921 ctccatcatg aagtgtgacg tggacatccg caaagacctg tacgccaaca cagtgctgtc
     1981 tggcggcacc accatgtacc ctggcattgc cgacaggatg cagaaggaga tcactgccct
     2041 ggcacccagc acaatgaaga tcaagatcat tgctcctcct gagcgcaagt actccgtgtg
     2101 gatcggcggc tccatcctgg cctcgctgtc caccttccag cagatgtgga tcagcaagca
     2161 ggagtatgac gagtccggcc cctccatcgt ccaccgcaaa tgcttcctcg agccaaccac
     2221 tgaggatctg tactttcaga gcgataacga tggatccgaa atcggtactg gctttccatt
     2281 cgacccccat tatgtggaag tcctgggcga gcgcatgcac tacgtcgatg ttggtccgcg
     2341 cgatggcacc cctgtgctgt tcctgcacgg taacccgacc tcctcctacg tgtggcgcaa
     2401 catcatcccg catgttgcac cgacccatcg ctgcattgct ccagacctga tcggtatggg
     2461 caaatccgac aaaccagacc tgggttattt cttcgacgac cacgtccgct tcatggatgc
     2521 cttcatcgaa gccctgggtc tggaagaggt cgtcctggtc attcacgact ggggctccgc
     2581 tctgggtttc cactgggcca agcgcaatcc agagcgcgtc aaaggtattg catttatgga
     2641 gttcatccgc cctatcccga cctgggacga atggccagaa tttgcccgcg agaccttcca
     2701 ggccttccgc accaccgacg tcggccgcaa gctgatcatc gatcagaacg tttttatcga
     2761 gggtacgctg ccgatgggtg tcgtccgccc gctgactgaa gtcgagatgg accattaccg
     2821 cgagccgttc ctgaatcctg ttgaccgcga gccactgtgg cgcttcccaa acgagctgcc
     2881 aatcgccggt gagccagcga acatcgtcgc gctggtcgaa gaatacatgg actggctgca
     2941 ccagtcccct gtcccgaagc tgctgttctg gggcacccca ggcgttctga tcccaccggc
     3001 cgaagccgct cgcctggcca aaagcctgcc taactgcaag gctgtggaca tcggcccggg
     3061 tctgaatctg ctgcaagaag acaacccgga cctgatcggc agcgagatcg cgcgctggct
     3121 gtctactctg gagatttccg gttaatagaa ttggcatgca agctgatccg gctgctaaca
     3181 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
     3241 ttggggcggc cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca
     3301 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     3361 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     3421 caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta caaatgtggt
     3481 aaaatcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt
     3541 acgcatctgt gcggtatttc acaccgcata cgcggatctg cgcagcacca tggcctgaaa
     3601 taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac cagctgtgga
     3661 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3721 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3781 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3841 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3901 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3961 gaggcttttt tggaggccta ggcttttgca aaaagcttga ttcttctgac acaacagtct
     4021 cgaacttaag gctagagcca ccatgattga acaagatgga ttgcacgcag gttctccggc
     4081 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4141 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4201 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     4261 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4321 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4381 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4441 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4501 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4561 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4621 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4681 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4741 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4801 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4861 accgaccaag cgacgcccaa cctgccatca cgatggccgc aataaaatat ctttattttc
     4921 attacatctg tgtgttggtt ttttgtgtga atcgatagcg ataaggatcc tctttgcgct
     4981 tgcgttttcc cttgtccaga tagcccagta gctgacattc atccggggtc agcaccgttt
     5041 ctgcggactg gctttctacg taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     5101 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5161 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     5221 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5281 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     5341 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     5401 tccaatgatg agcactttca aagttctgct atgtggcgcg gtattatccc gtattgacgc
     5461 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     5521 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     5581 cataaccatg agtgataaca ctgcggccaa cttacttctg acaactatcg gaggaccgaa
     5641 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     5701 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     5761 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5821 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5881 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5941 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     6001 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     6061 gcattggtaa ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc tcactcaaag
     6121 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6181 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6241 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6301 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6361 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6421 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6481 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6541 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6601 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     6661 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     6721 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     6781 aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag atcctttgat cttttctacg
     6841 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6901 aaaaggatct tcacctagat ccttttatag tccggaaata caggaacgca cgctggatgg
     6961 cccttcgctg ggatggtgaa accatgaaaa atggcagctt cagtggatta agtgggggta
     7021 atgtggcctg taccctctgg ttgcataggt attcatacgg ttaaaattta tcaggcgcga
     7081 ttgcggcagt ttttcgggtg gtttgttgcc atttttacct gtctgctgcc gtgatcgcgc
     7141 tgaacgcgtt ttagcggtgc gtacaattaa gggattatgg taaatccact tactgtctgc
     7201 cctcgtagcc atcgagataa accgcagtac tccggccacg atgcgtccgg cgtagaggat
     7261 cgagatct
//
