LOCUS       sequence-426151-        6617 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect expression of human EML4 fragment I250 to A877. N-terminal 6X
            His tag with TEV cleavage..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6617)
  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 6617)
  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_EML4:250-877
FEATURES             Location/Qualifiers
     source          1..6617
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1292..1426
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1329..1348)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1383..1402
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1455..1620
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      1804..2258
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     2100..2121
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2284..2388
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2389..2457,2458..3249)
                     /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(2607..2626)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3420..4008
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3909..3928
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4162..4179
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  4313..4537
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4604..5137)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5307..5312,5313..5329,5330..5335))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        5687..5778
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     5703..5720
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             5837..5854
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5867..5887
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 aaggacctaa aggtgtatat caaatcagca aacaaatcaa agctcatgat ggcagtgtgt
       61 tcacactttg tcagatgaga aatgggatgt tattaactgg aggagggaaa gacagaaaaa
      121 taattctgtg ggatcatgat ctgaatcctg aaagagaaat agaggttcct gatcagtatg
      181 gcacaatcag agctgtagca gaaggaaagg cagatcaatt tttagtaggc acatcacgaa
      241 actttatttt acgaggaaca tttaatgatg gcttccaaat agaagtacag ggtcatacag
      301 atgagctttg gggtcttgcc acacatccct tcaaagattt gctcttgaca tgtgctcagg
      361 acaggcaggt gtgcctgtgg aactcaatgg aacacaggct ggaatggacc aggctggtag
      421 atgaaccagg acactgtgca gattttcatc caagtggcac agtggtggcc ataggaacgc
      481 actcaggcag gtggtttgtt ctggatgcag aaaccagaga tctagtttct atccacacag
      541 acgggaatga acagctctct gtgatgcgct actcaataga tggtaccttc ctggctgtag
      601 gatctcatga caactttatt tacctctatg tagtctctga aaatggaaga aaatatagca
      661 gatatggaag gtgcactgga cattccagct acatcacaca ccttgactgg tccccagaca
      721 acaagtatat aatgtctaac tcgggagact atgaaatatt gtactgggac attccaaatg
      781 gctgcaaact aatcaggaat cgatcggatt gtaaggacat tgattggacg acatatacct
      841 gtgtgctagg atttcaagta tttggtgtct ggccagaagg atctgatggg acagatatca
      901 atgcactggt gcgatcccac aatagaaagg tgatagctgt tgccgatgac ttttgtaaag
      961 tccatctgtt tcagtatccc tgctccaaag caaaggctcc cagtcacaag tacagtgccc
     1021 acagcagcca tgtcaccaat gtcagtttta ctcacaatga cagtcacctg atatcaactg
     1081 gtggaaaaga catgagcatc attcagtgga aacttgtgga aaagttatct ttgcctcaga
     1141 atgagactgt agcgtgatga cgaagcttgt cgagaagtac tagaggatca taatcagcca
     1201 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     1261 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     1321 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     1381 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatctgat cactgatatc
     1441 gcctaggaga tccgaaccag ataagtgaaa tctagttcca aactattttg tcatttttaa
     1501 ttttcgtatt agcttacgac gctacaccca gttcccatct attttgtcac tcttccctaa
     1561 ataatcctta aaaactccat ttccacccct cccagttccc aactattttg tccgcccaca
     1621 gcggggcatt tttcttcctg ttatgttttt aatcaaacat cctgccaact ccatgtgaca
     1681 aaccgtcatc ttcggctact ttttctctgt cacagaatga aaatttttct gtcatctctt
     1741 cgttattaat gtttgtaatt gactgaatat caacgcttat ttgcagcctg aatggcgaat
     1801 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     1861 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     1921 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     1981 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     2041 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttcttaatag
     2101 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     2161 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2221 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     2281 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2341 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2401 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2461 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2521 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2581 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     2641 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2701 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2761 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     2821 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     2881 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     2941 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3001 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3061 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3121 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3181 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3241 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3301 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3361 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3421 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3481 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3541 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     3601 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3661 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3721 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3781 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3841 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3901 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3961 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4021 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4081 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4141 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     4201 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagacca gccgcgtaac
     4261 ctggcaaaat cggttacggt tgagtaataa atggatgccc tgcgtaagcg ggtgtgggcg
     4321 gacaataaag tcttaaactg aacaaaatag atctaaacta tgacaataaa gtcttaaact
     4381 agacagaata gttgtaaact gaaatcagtc cagttatgct gtgaaaaagc atactggact
     4441 tttgttatgg ctaaagcaaa ctcttcattt tctgaagtgc aaattgcccg tcgtattaaa
     4501 gaggggcgtg gccaagggca tggtaaagac tatattcgcg gcgttgtgac aatttaccga
     4561 acaactccgc ggccgggaag ccgatctcgg cttgaacgaa ttgttaggtg gcggtacttg
     4621 ggtcgatatc aaagtgcatc acttcttccc gtatgcccaa ctttgtatag agagccactg
     4681 cgggatcgtc accgtaatct gcttgcacgt agatcacata agcaccaagc gcgttggcct
     4741 catgcttgag cagattgatg agcgcggtgg caatgccctg cctccggtgc tcgccggaga
     4801 ctgcgagatc atagatatag atctcactac gcggctgctc aaacttgggc agaacgtaag
     4861 ccgcgagagc gccaacaacc gcttcttggt cgaaggcagc aagcgcgatg aatgtcttac
     4921 tacggagcaa gttcccgagg taatcggagt ccggctgatg ttgggagtag gtggctacgt
     4981 ctccgaactc acgaccgaaa agatcaagag cagcccgcat ggatttgact tggtcagggc
     5041 cgagcctaca tgtgcgaatg atgcccatac ttgagccacc taactttgtt ttagggcgac
     5101 tgccctgctg cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc acggcgtaac
     5161 gcgcttgctg cttggatgcc cgaggcatag actgtacaaa aaaacagtca taacaagcca
     5221 tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt caaggttctg gaccagttgc
     5281 gtgagcgcat acgctacttg cattacagtt tacgaaccga acaggcttat gtcaactggg
     5341 ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc aaccttgggc agcagcgaag
     5401 tcgaggcatt tctgtcctgg ctggcgaacg agcgcaaggt ttcggtctcc acgcatcgtc
     5461 aggcattggc ggccttgctg ttcttctacg gcaaggtgct gtgcacggat ctgccctggc
     5521 ttcaggagat cggaagacct cggccgtcgc ggcgcttgcc ggtggtgctg accccggatg
     5581 aagtggttcg catcctcggt tttctggaag gcgagcatcg tttgttcgcc caggactcta
     5641 gctatagttc tagtggttgg ctacgtatac tccggaatat taatagatca tggagataat
     5701 taaaatgata accatctcgc aaataaataa gtattttact gttttcgtaa cagttttgta
     5761 ataaaaaaac ctataaatat tccggattat tcataccgtc ccaccatcgg gcgcggatct
     5821 cggtccgaaa accatgcatc atcatcatca tcacagcagc ggcagagaaa acttgtattt
     5881 ccagggcatc agaacggaac tgcctcctga gaagctcaaa ctggagtggg catatggtta
     5941 tcgaggaaag gactgtagag ctaatgttta ccttcttccg accggggaaa tagtttattt
     6001 cattgcatca gtagtagtac tatttaatta tgaggagaga actcagcgac actacctggg
     6061 ccatacagac tgtgtgaaat gccttgctat acatcctgac aaaattagga ttgcaactgg
     6121 acagatagct ggcgtggata aagatggaag gcctctacaa ccccacgtca gagtgtggga
     6181 ttctgttact ctatccacac tgcagattat tggacttggc acttttgagc gtggagtagg
     6241 atgcctggat ttttcaaaag cagattcagg tgttcattta tgtgttattg atgactccaa
     6301 tgagcatatg cttactgtat gggactggca gaagaaagca aaaggagcag aaataaagac
     6361 aacaaatgaa gttgttttgg ctgtggagtt tcacccaaca gatgcaaata ccataattac
     6421 atgcggtaaa tctcatattt tcttctggac ctggagcggc aattcactaa caagaaaaca
     6481 gggaattttt gggaaatatg aaaagccaaa atttgtgcag tgtttagcat tcttggggaa
     6541 tggagatgtt cttactggag actcaggtgg agtcatgctt atatggagca aaactactgt
     6601 agagcccaca cctggga
//
