LOCUS       sequence-423308-        7256 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human ACTB. N-terminal HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7256)
  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 7256)
  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: pHTN-HaloTag_ACTB:1-375
FEATURES             Location/Qualifiers
     source          1..7256
                     /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"
     regulatory      1061..1070
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1067..1957
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     CDS             1970..1990
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     primer_bind     complement(3207..3225)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3257..3378
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3268..3287)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3322..3341
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3623..3980
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3623..3814
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3646..3666)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3831..3966
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4031..4825
                     /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(4085..4104)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4695..4714
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4889..4937
                     /note="synthetic polyadenylation signal"
     promoter        5093..5197
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5198..5266,5267..6058)
                     /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(5416..5435)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6216..6804)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6296..6315)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6920..7203
                     /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 gccaccatgg cagaaatcgg
     1081 tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca tgcactacgt
     1141 cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc cgacctcctc
     1201 ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca ttgctccaga
     1261 cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg acgaccacgt
     1321 ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc tggtcattca
     1381 cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc gcgtcaaagg
     1441 tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc cagaatttgc
     1501 ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga tcatcgatca
     1561 gaacgttttt atcgagggta cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga
     1621 gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt
     1681 cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata
     1741 catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca ccccaggcgt
     1801 tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt
     1861 ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga tcggcagcga
     1921 gatcgcgcgc tggctgtcga cgctcgagat ttccggcgag ccaaccactg aggatctgta
     1981 ctttcagagc gataacgcga tcgcttccga attcatggat gatgatatcg ccgcgctcgt
     2041 cgtcgacaac ggctccggca tgtgcaaggc cggcttcgcg ggcgacgatg ccccccgggc
     2101 cgtcttcccc tccatcgtgg ggcgccccag gcaccagggc gtgatggtgg gcatgggtca
     2161 gaaggattcc tatgtgggcg acgaggccca gagcaagaga ggcatcctca ccctgaagta
     2221 ccccatcgag cacggcatcg tcaccaactg ggacgacatg gagaaaatct ggcaccacac
     2281 cttctacaat gagctgcgtg tggctcccga ggagcacccc gtgctgctga ccgaggcccc
     2341 cctgaacccc aaggccaacc gcgagaagat gacccagatc atgtttgaga ccttcaacac
     2401 cccagccatg tacgttgcta tccaggctgt gctatccctg tacgcctctg gccgtaccac
     2461 tggcatcgtg atggactccg gtgacggggt cacccacact gtgcccatct acgaggggta
     2521 tgccctcccc catgccatcc tgcgtctgga cctggctggc cgggacctga ctgactacct
     2581 catgaagatc ctcaccgagc gcggctacag cttcaccacc acggccgagc gggaaatcgt
     2641 gcgtgacatt aaggagaagc tgtgctacgt cgccctggac ttcgagcaag agatggccac
     2701 ggctgcttcc agctcctccc tggagaagag ctacgagctg cctgacggcc aggtcatcac
     2761 cattggcaat gagcggttcc gctgccctga ggcactcttc cagccttcct tcctgggcat
     2821 ggagtcctgt ggcatccacg aaactacctt caactccatc atgaagtgtg acgtggacat
     2881 ccgcaaagac ctgtacgcca acacagtgct gtctggcggc accaccatgt accctggcat
     2941 tgccgacagg atgcagaagg agatcactgc cctggcaccc agcacaatga agatcaagat
     3001 cattgctcct cctgagcgca agtactccgt gtggatcggc ggctccatcc tggcctcgct
     3061 gtccaccttc cagcagatgt ggatcagcaa gcaggagtat gacgagtccg gcccctccat
     3121 cgtccaccgc aaatgcttct gatgatctag atttgggccc aattcctgca ggattttgcg
     3181 gccgcttgct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggccg
     3241 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     3301 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     3361 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     3421 gagatgtggg aggttttttt aagcaagtaa aacctctaca aatgtggtaa aatcgaattt
     3481 taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc
     3541 ggtatttcac accgcatacg cggatctgcg cagcaccatg gcctgaaata acctctgaaa
     3601 gaggaacttg gttaggtacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     3661 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3721 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3781 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3841 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3901 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3961 gaggcctagg cttttgcaaa aagcttgatt cttctgacac aacagtctcg aacttaaggc
     4021 tagagccacc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4081 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4141 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4201 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4261 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4321 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4381 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4441 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4501 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     4561 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4621 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4681 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4741 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4801 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4861 acgcccaacc tgccatcacg atggccgcaa taaaatatct ttattttcat tacatctgtg
     4921 tgttggtttt ttgtgtgaat cgatagcgat aaggatcctc tttgcgcttg cgttttccct
     4981 tgtccagata gcccagtagc tgacattcat ccggggtcag caccgtttct gcggactggc
     5041 tttctacgta atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     5101 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5161 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5221 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5281 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5341 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5401 cactttcaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5461 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5521 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5581 tgataacact gcggccaact tacttctgac aactatcgga ggaccgaagg agctaaccgc
     5641 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5701 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5761 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5821 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5881 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5941 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6001 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaatt
     6061 cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg
     6121 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     6181 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6241 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6301 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6361 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6421 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6481 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6541 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6601 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     6661 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6721 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6781 acgcgcagaa aaaaaggatt tcaagaagat cctttgatct tttctacggg gtctgacgct
     6841 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6901 acctagatcc ttttatagtc cggaaataca ggaacgcacg ctggatggcc cttcgctggg
     6961 atggtgaaac catgaaaaat ggcagcttca gtggattaag tgggggtaat gtggcctgta
     7021 ccctctggtt gcataggtat tcatacggtt aaaatttatc aggcgcgatt gcggcagttt
     7081 ttcgggtggt ttgttgccat ttttacctgt ctgctgccgt gatcgcgctg aacgcgtttt
     7141 agcggtgcgt acaattaagg gattatggta aatccactta ctgtctgccc tcgtagccat
     7201 cgagataaac cgcagtactc cggccacgat gcgtccggcg tagaggatcg agatct
//
