LOCUS       sequence-424690-        7259 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human TRIB1. C-terminal 
            HaloTag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7259)
  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 7259)
  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_TRIB1:1-372
FEATURES             Location/Qualifiers
     source          1..7259
                     /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             2213..2233
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2249..3133
                     /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(3205..3223)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3260..3381
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3271..3290)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3325..3344
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3626..3983
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3626..3817
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3649..3669)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3834..3969
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4034..4828
                     /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(4088..4107)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4698..4717
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4892..4940
                     /note="synthetic polyadenylation signal"
     promoter        5096..5200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5201..5269,5270..6061)
                     /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(5419..5438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6219..6807)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6299..6318)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6923..7206
                     /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 catgcgagtc ggtccggtgc gctctgccat gagcggcgcc tcgcagcccc gcggcccggc
     1141 cctgctcttc ccagccaccc gaggcgtccc ggccaaacgc ctgctggacg ccgacgacgc
     1201 ggcggctgtg gcggccaagt gcccgcgcct ctccgagtgc tccagccccc cggactacct
     1261 cagccccccc ggctcgccct gcagtccgca gcccccgcct gccgctccgg gggccggcgg
     1321 aggctccggg agcgcgccgg ggcccagccg catcgccgac tacctgctgc tgcccctagc
     1381 cgagcgcgag catgtgtccc gggcgctgtg catccacact ggacgcgagc tgcgctgcaa
     1441 ggtgtttccc attaaacact accaggacaa aatcaggcct tacatccagc tgccatcgca
     1501 cagcaacatt actggcattg tggaagtgat ccttggggaa accaaggcct atgtcttctt
     1561 tgagaaggac tttggggaca tgcactccta tgtgcgaagc cggaagaggc tgcgggaaga
     1621 ggaagccgcc cggctcttca agcagattgt ctccgccgtc gcccactgcc accagtcagc
     1681 catcgtgctg ggggacctga agcttaggaa gttcgtcttc tccacggagg agagaaccca
     1741 gcttagacta gaaagtctag aagacacaca cataatgaag ggggaagatg atgctttgtc
     1801 agacaaacat ggctgcccag cctacgtgag ccctgagatc ctcaacacca ctgggaccta
     1861 ctccggaaag gctgcggacg tttggagcct gggggtgatg ctctacaccc ttctggttgg
     1921 acgatacccc ttccatgact cagaccccag tgcccttttc tccaaaattc ggcgtggaca
     1981 gttctgcatt cctgagcaca tttcccccaa agccaggtgc ctcattcgca gcctcttgag
     2041 acgggagccc tccgagagac tcactgcccc cgagatccta ctgcacccct ggtttgagtc
     2101 cgtcttggaa cccgggtaca tcgactcaga aataggaact tcagaccaga ttgttccaga
     2161 gtaccaggag gacagtgaca ttagttcctt cttctgcctc gagccaacca ctgaggatct
     2221 gtactttcag agcgataacg atggatccga aatcggtact ggctttccat tcgaccccca
     2281 ttatgtggaa gtcctgggcg agcgcatgca ctacgtcgat gttggtccgc gcgatggcac
     2341 ccctgtgctg ttcctgcacg gtaacccgac ctcctcctac gtgtggcgca acatcatccc
     2401 gcatgttgca ccgacccatc gctgcattgc tccagacctg atcggtatgg gcaaatccga
     2461 caaaccagac ctgggttatt tcttcgacga ccacgtccgc ttcatggatg ccttcatcga
     2521 agccctgggt ctggaagagg tcgtcctggt cattcacgac tggggctccg ctctgggttt
     2581 ccactgggcc aagcgcaatc cagagcgcgt caaaggtatt gcatttatgg agttcatccg
     2641 ccctatcccg acctgggacg aatggccaga atttgcccgc gagaccttcc aggccttccg
     2701 caccaccgac gtcggccgca agctgatcat cgatcagaac gtttttatcg agggtacgct
     2761 gccgatgggt gtcgtccgcc cgctgactga agtcgagatg gaccattacc gcgagccgtt
     2821 cctgaatcct gttgaccgcg agccactgtg gcgcttccca aacgagctgc caatcgccgg
     2881 tgagccagcg aacatcgtcg cgctggtcga agaatacatg gactggctgc accagtcccc
     2941 tgtcccgaag ctgctgttct ggggcacccc aggcgttctg atcccaccgg ccgaagccgc
     3001 tcgcctggcc aaaagcctgc ctaactgcaa ggctgtggac atcggcccgg gtctgaatct
     3061 gctgcaagaa gacaacccgg acctgatcgg cagcgagatc gcgcgctggc tgtctactct
     3121 ggagatttcc ggttaataga attggcatgc aagctgatcc ggctgctaac aaagcccgaa
     3181 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcgg
     3241 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     3301 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     3361 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     3421 ggggagatgt gggaggtttt tttaagcaag taaaacctct acaaatgtgg taaaatcgaa
     3481 ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg
     3541 tgcggtattt cacaccgcat acgcggatct gcgcagcacc atggcctgaa ataacctctg
     3601 aaagaggaac ttggttaggt accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc
     3661 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3721 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3781 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3841 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3901 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     3961 ttggaggcct aggcttttgc aaaaagcttg attcttctga cacaacagtc tcgaacttaa
     4021 ggctagagcc accatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4081 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4141 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4201 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4261 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4321 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4381 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4441 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4501 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4561 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4621 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4681 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4741 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4801 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     4861 gcgacgccca acctgccatc acgatggccg caataaaata tctttatttt cattacatct
     4921 gtgtgttggt tttttgtgtg aatcgatagc gataaggatc ctctttgcgc ttgcgttttc
     4981 ccttgtccag atagcccagt agctgacatt catccggggt cagcaccgtt tctgcggact
     5041 ggctttctac gtaatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     5101 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     5161 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     5221 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     5281 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     5341 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     5401 gagcactttc aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     5461 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     5521 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     5581 gagtgataac actgcggcca acttacttct gacaactatc ggaggaccga aggagctaac
     5641 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     5701 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     5761 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     5821 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5881 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5941 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     6001 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     6061 attcgaaatg accgaccaag cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata
     6121 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6181 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6241 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6301 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6361 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6421 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6481 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6541 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6601 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     6661 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6721 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6781 attacgcgca gaaaaaaagg atttcaagaa gatcctttga tcttttctac ggggtctgac
     6841 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6901 ttcacctaga tccttttata gtccggaaat acaggaacgc acgctggatg gcccttcgct
     6961 gggatggtga aaccatgaaa aatggcagct tcagtggatt aagtgggggt aatgtggcct
     7021 gtaccctctg gttgcatagg tattcatacg gttaaaattt atcaggcgcg attgcggcag
     7081 tttttcgggt ggtttgttgc catttttacc tgtctgctgc cgtgatcgcg ctgaacgcgt
     7141 tttagcggtg cgtacaatta agggattatg gtaaatccac ttactgtctg ccctcgtagc
     7201 catcgagata aaccgcagta ctccggccac gatgcgtccg gcgtagagga tcgagatct
//
