LOCUS       sequence-426220-        5633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect expression of human TLE4 fragment I474 to Y773. N-terminal 6X
            His tag with TEV cleavage..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5633)
  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 5633)
  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: pFBOH-MHL_TLE4:474-773
FEATURES             Location/Qualifiers
     source          1..5633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  75..299
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(366..899)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1069..1074,1075..1091,1092..1097))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        1449..1540
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1465..1482
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             1599..1616
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1629..1649
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    2687..2821
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2724..2743)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2778..2797
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2850..3015
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3199..3653
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3286..3305)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3495..3516
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3679..3783
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3784..3852,3853..4644)
                     /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(4002..4021)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4815..5403
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5304..5323
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5557..5574
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ccgcatagac cagccgcgta acctggcaaa atcggttacg gttgagtaat aaatggatgc
       61 cctgcgtaag cgggtgtggg cggacaataa agtcttaaac tgaacaaaat agatctaaac
      121 tatgacaata aagtcttaaa ctagacagaa tagttgtaaa ctgaaatcag tccagttatg
      181 ctgtgaaaaa gcatactgga cttttgttat ggctaaagca aactcttcat tttctgaagt
      241 gcaaattgcc cgtcgtatta aagaggggcg tggccaaggg catggtaaag actatattcg
      301 cggcgttgtg acaatttacc gaacaactcc gcggccggga agccgatctc ggcttgaacg
      361 aattgttagg tggcggtact tgggtcgata tcaaagtgca tcacttcttc ccgtatgccc
      421 aactttgtat agagagccac tgcgggatcg tcaccgtaat ctgcttgcac gtagatcaca
      481 taagcaccaa gcgcgttggc ctcatgcttg agcagattga tgagcgcggt ggcaatgccc
      541 tgcctccggt gctcgccgga gactgcgaga tcatagatat agatctcact acgcggctgc
      601 tcaaacttgg gcagaacgta agccgcgaga gcgccaacaa ccgcttcttg gtcgaaggca
      661 gcaagcgcga tgaatgtctt actacggagc aagttcccga ggtaatcgga gtccggctga
      721 tgttgggagt aggtggctac gtctccgaac tcacgaccga aaagatcaag agcagcccgc
      781 atggatttga cttggtcagg gccgagccta catgtgcgaa tgatgcccat acttgagcca
      841 cctaactttg ttttagggcg actgccctgc tgcgtaacat cgttgctgct ccataacatc
      901 aaacatcgac ccacggcgta acgcgcttgc tgcttggatg cccgaggcat agactgtaca
      961 aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc gctgcgttcg
     1021 gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag tttacgaacc
     1081 gaacaggctt atgtcaactg ggttcgtgcc ttcatccgtt tccacggtgt gcgtcacccg
     1141 gcaaccttgg gcagcagcga agtcgaggca tttctgtcct ggctggcgaa cgagcgcaag
     1201 gtttcggtct ccacgcatcg tcaggcattg gcggccttgc tgttcttcta cggcaaggtg
     1261 ctgtgcacgg atctgccctg gcttcaggag atcggaagac ctcggccgtc gcggcgcttg
     1321 ccggtggtgc tgaccccgga tgaagtggtt cgcatcctcg gttttctgga aggcgagcat
     1381 cgtttgttcg cccaggactc tagctatagt tctagtggtt ggctacgtat actccggaat
     1441 attaatagat catggagata attaaaatga taaccatctc gcaaataaat aagtatttta
     1501 ctgttttcgt aacagttttg taataaaaaa acctataaat attccggatt attcataccg
     1561 tcccaccatc gggcgcggat ctcggtccga aaaccatgca tcatcatcat catcacagca
     1621 gcggcagaga aaacttgtat ttccagggca tcccccggca tgctcgccag atcaacaccc
     1681 tcaaccacgg ggaggtggtg tgcgcggtga ccatcagcaa ccccacgaga cacgtgtaca
     1741 cgggtgggaa gggctgcgtc aaggtctggg acatcagcca cccaggcaat aagagtcctg
     1801 tctcccagct cgactgtctg aacagggata actacatccg ttcctgcaga ttgctccctg
     1861 atggtcgcac cctaattgtt ggaggggaag ccagtacttt gtccatttgg gacctggcgg
     1921 ctccaacccc acgcatcaag gcagagctga catcctcggc ccccgcctgc tatgccctgg
     1981 ccatcagccc cgattccaag gtctgcttct catgctgcag cgacggcaac atcgctgtgt
     2041 gggatctgca caaccagacc ttggtgaggc aattccaggg ccacacagat ggagccagct
     2101 gtattgacat ttctaatgat ggcaccaagc tctggacagg tggtttggac aacacggtca
     2161 ggtcctggga cctgcgcgag gggcggcagc tgcagcagca cgacttcacc tcccagatct
     2221 tttctctggg ctactgccca actggagagt ggcttgcagt ggggatggag aacagcaatg
     2281 tggaagtttt gcatgtcacc aagccagaca aataccaact acatcttcat gagagctgtg
     2341 tgctgtcgct caagtttgcc cattgtggca aatggtttgt aagcactgga aaggacaacc
     2401 ttctgaatgc ctggagaaca ccttatgggg ccagtatatt ccagtccaaa gaatcctcat
     2461 cggtgcttag ctgtgacatc tccgtggacg acaaatacat tgtcactggc tctggggata
     2521 agaaggccac agtttatgaa gttatttatt gatgacgaag cttgtcgaga agtactagag
     2581 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     2641 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     2701 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     2761 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     2821 ctgatcactg atatcgccta ggagatccga accagataag tgaaatctag ttccaaacta
     2881 ttttgtcatt tttaattttc gtattagctt acgacgctac acccagttcc catctatttt
     2941 gtcactcttc cctaaataat ccttaaaaac tccatttcca cccctcccag ttcccaacta
     3001 ttttgtccgc ccacagcggg gcatttttct tcctgttatg tttttaatca aacatcctgc
     3061 caactccatg tgacaaaccg tcatcttcgg ctactttttc tctgtcacag aatgaaaatt
     3121 tttctgtcat ctcttcgtta ttaatgtttg taattgactg aatatcaacg cttatttgca
     3181 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3241 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     3301 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3361 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     3421 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3481 ccacgttctt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3541 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3601 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     3661 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3721 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3781 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3841 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3901 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3961 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     4021 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     4081 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     4141 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4201 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4261 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4321 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4381 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4441 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4501 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4561 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4621 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4681 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4741 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4801 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4861 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4921 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     4981 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     5041 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     5101 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     5161 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5221 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5281 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5341 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5401 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5461 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     5521 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     5581 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc aca
//
