LOCUS       sequence-423371-        8189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human MTA2. C-terminal HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8189)
  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 8189)
  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_MTA2:1-668
FEATURES             Location/Qualifiers
     source          1..8189
                     /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      1079..1088
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1083..1092
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3143..3163
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             3179..4063
                     /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(4135..4153)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4190..4311
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4201..4220)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4255..4274
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4556..4913
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4556..4747
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4579..4599)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4764..4899
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4964..5758
                     /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(5018..5037)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5628..5647
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5822..5870
                     /note="synthetic polyadenylation signal"
     promoter        6026..6130
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6131..6199,6200..6991)
                     /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(6349..6368)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7149..7737)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7229..7248)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7853..8136
                     /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 ccgaattcgc
     1081 caccatggcg gccaacatgt accgggtggg agattacgtc tattttgaga actcttccag
     1141 caatccttac ctggttagac ggattgagga gctcaacaag actgcaaatg gaaatgtgga
     1201 ggcaaaggtt gtctgtcttt tccggcgcag ggacatttct agtagcctca acagcctggc
     1261 tgatagtaat gccagggagt ttgaagagga atcaaagcag ccaggggtgt ctgagcagca
     1321 gcgccatcaa ctgaagcacc gggaactttt tctttctcgg caatttgaat cattaccagc
     1381 cacccacata cgggggaaat gcagtgtgac cctcttgaat gagacagata tcttgagcca
     1441 gtacctggaa aaggaggact gcttttttta ctcactggtg tttgaccccg tgcagaagac
     1501 acttctcgct gatcagggcg agattagagt tggttgcaaa taccaagctg agatcccaga
     1561 tcgcctagta gagggagaat ctgataatcg gaaccagcag aagatggaga tgaaggtctg
     1621 ggacccagac aaccctctca cagaccggca gatcgaccag tttcttgtgg tggcccgagc
     1681 tgtgggaacc tttgcaagag ccctagattg tagcagctcc attcggcagc caagcttgca
     1741 catgagtgca gctgctgcct cccgagatat cactctgttt cacgccatgg ataccttgca
     1801 aaggaacggc tacgacctgg ctaaggccat gtcgaccctg gtaccccagg gaggcccggt
     1861 gctgtgtcgg gatgagatgg aggaatggtc agcctcagag gccatgctat ttgaggaggc
     1921 cctagagaag tatgggaagg acttcaatga tattcgccag gattttctac cctggaagtc
     1981 acttgccagc atagtccagt tttattacat gtggaaaacc acagaccggt atattcagca
     2041 gaaaaggttg aaagctgctg aagcagacag caaactgaaa caggtctaca ttcccaccta
     2101 cactaagcca aaccctaacc agatcatttc tgtgggttca aaacctggca tgaatggggc
     2161 tggatttcag aagggcctga cttgtgagag ttgccacacc acacagtctg ctcagtggta
     2221 tgcctggggc ccacctaaca tgcagtgccg cctctgtgct tcctgttgga tctactggaa
     2281 gaagtatggg ggactgaaga ccccaactca gcttgagggg gccactcggg gcaccacgga
     2341 gccacactca aggggtcatt tatccagacc tgaagctcaa agtctctctc cttacacaac
     2401 cagcgccaac agggccaagc tactggctaa gaacagacaa actttcctgc ttcagaccac
     2461 aaagctgacc cgtcttgcca gacgcatgtg cagggaccta ttacagccaa ggagggccgc
     2521 ccgacggcct tatgctccta tcaatgccaa tgccatcaaa gcagagtgct ccattcgact
     2581 tcctaaggcc gccaagactc cattgaagat tcaccctctg gtgcggctgc ccctggcaac
     2641 tatcgtcaaa gatctggtgg cccaggcacc cctgaaacca aaaacacctc ggggtaccaa
     2701 gacaccgatc aacagaaacc agctgtccca gaaccgggga ctggggggca ttatggtgaa
     2761 acgggcctat gagactatgg caggggcagg ggttcctttc tctgccaatg gaaggcctct
     2821 ggcttcaggg attcgttcaa gctcacagcc agcagccaag cgtcagaaac taaacccagc
     2881 tgatgccccc aatcctgtgg tgtttgtggc cacaaaggat accagggccc tacggaaggc
     2941 tctgacccat ctggaaatgc ggcgagctgc tcgccgaccc aacttgcccc tgaaggtgaa
     3001 gccaacgctg attgcagtgc ggccccctgt ccctctacct gcaccctcac atcctgccag
     3061 caccaatgag cctattgtcc tggaggaccg gatatctaga tttgggccca attcctgcag
     3121 gcgagctctc gagccaacca ctgaggatct gtactttcag agcgataacg atggatccga
     3181 aatcggtact ggctttccat tcgaccccca ttatgtggaa gtcctgggcg agcgcatgca
     3241 ctacgtcgat gttggtccgc gcgatggcac ccctgtgctg ttcctgcacg gtaacccgac
     3301 ctcctcctac gtgtggcgca acatcatccc gcatgttgca ccgacccatc gctgcattgc
     3361 tccagacctg atcggtatgg gcaaatccga caaaccagac ctgggttatt tcttcgacga
     3421 ccacgtccgc ttcatggatg ccttcatcga agccctgggt ctggaagagg tcgtcctggt
     3481 cattcacgac tggggctccg ctctgggttt ccactgggcc aagcgcaatc cagagcgcgt
     3541 caaaggtatt gcatttatgg agttcatccg ccctatcccg acctgggacg aatggccaga
     3601 atttgcccgc gagaccttcc aggccttccg caccaccgac gtcggccgca agctgatcat
     3661 cgatcagaac gtttttatcg agggtacgct gccgatgggt gtcgtccgcc cgctgactga
     3721 agtcgagatg gaccattacc gcgagccgtt cctgaatcct gttgaccgcg agccactgtg
     3781 gcgcttccca aacgagctgc caatcgccgg tgagccagcg aacatcgtcg cgctggtcga
     3841 agaatacatg gactggctgc accagtcccc tgtcccgaag ctgctgttct ggggcacccc
     3901 aggcgttctg atcccaccgg ccgaagccgc tcgcctggcc aaaagcctgc ctaactgcaa
     3961 ggctgtggac atcggcccgg gtctgaatct gctgcaagaa gacaacccgg acctgatcgg
     4021 cagcgagatc gcgcgctggc tgtctactct ggagatttcc ggttaataga attggcatgc
     4081 aagctgatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
     4141 agcaataact agcataaccc cttggggcgg ccgcttcgag cagacatgat aagatacatt
     4201 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     4261 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     4321 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt tttaagcaag
     4381 taaaacctct acaaatgtgg taaaatcgaa ttttaacaaa atattaacgc ttacaatttc
     4441 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgcggatct
     4501 gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt accttctgag
     4561 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4621 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4681 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4741 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4801 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     4861 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagcttg
     4921 attcttctga cacaacagtc tcgaacttaa ggctagagcc accatgattg aacaagatgg
     4981 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     5041 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     5101 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     5161 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     5221 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     5281 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     5341 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     5401 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     5461 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     5521 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     5581 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     5641 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5701 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     5761 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgatggccg
     5821 caataaaata tctttatttt cattacatct gtgtgttggt tttttgtgtg aatcgatagc
     5881 gataaggatc ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt
     5941 catccggggt cagcaccgtt tctgcggact ggctttctac gtaatggttt cttagacgtc
     6001 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     6061 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6121 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     6181 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     6241 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     6301 ttttcgcccc gaagaacgtt ttccaatgat gagcactttc aaagttctgc tatgtggcgc
     6361 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     6421 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     6481 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     6541 gacaactatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     6601 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     6661 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6721 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6781 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6841 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6901 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6961 gataggtgcc tcactgatta agcattggta attcgaaatg accgaccaag cgacgcccaa
     7021 ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     7081 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     7141 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     7201 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     7261 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     7321 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     7381 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     7441 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     7501 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     7561 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     7621 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7681 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atttcaagaa
     7741 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7801 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttata gtccggaaat
     7861 acaggaacgc acgctggatg gcccttcgct gggatggtga aaccatgaaa aatggcagct
     7921 tcagtggatt aagtgggggt aatgtggcct gtaccctctg gttgcatagg tattcatacg
     7981 gttaaaattt atcaggcgcg attgcggcag tttttcgggt ggtttgttgc catttttacc
     8041 tgtctgctgc cgtgatcgcg ctgaacgcgt tttagcggtg cgtacaatta agggattatg
     8101 gtaaatccac ttactgtctg ccctcgtagc catcgagata aaccgcagta ctccggccac
     8161 gatgcgtccg gcgtagagga tcgagatct
//
