LOCUS       sequence-426146-        6803 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect expression of human EIF3B fragment D127 to E814. N-terminal 
            6X His tag with TEV cleavage..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6803)
  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 6803)
  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_EIF3B:127-814
FEATURES             Location/Qualifiers
     source          1..6803
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(484..489,490..506,507..512))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        864..955
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     880..897
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             1014..1031
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1044..1064
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    3272..3406
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3309..3328)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3363..3382
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3435..3600
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3784..4238
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3871..3890)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4080..4101
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4264..4368
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4369..4437,4438..5229)
                     /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(4587..4606)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5400..5988
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5889..5908
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6142..6159
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6293..6517
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(join(6584..6803,1..314))
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 tcactacgcg gctgctcaaa cttgggcaga acgtaagccg cgagagcgcc aacaaccgct
       61 tcttggtcga aggcagcaag cgcgatgaat gtcttactac ggagcaagtt cccgaggtaa
      121 tcggagtccg gctgatgttg ggagtaggtg gctacgtctc cgaactcacg accgaaaaga
      181 tcaagagcag cccgcatgga tttgacttgg tcagggccga gcctacatgt gcgaatgatg
      241 cccatacttg agccacctaa ctttgtttta gggcgactgc cctgctgcgt aacatcgttg
      301 ctgctccata acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga
      361 ggcatagact gtacaaaaaa acagtcataa caagccatga aaaccgccac tgcgccgtta
      421 ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg agcgcatacg ctacttgcat
      481 tacagtttac gaaccgaaca ggcttatgtc aactgggttc gtgccttcat ccgtttccac
      541 ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg aggcatttct gtcctggctg
      601 gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg cattggcggc cttgctgttc
      661 ttctacggca aggtgctgtg cacggatctg ccctggcttc aggagatcgg aagacctcgg
      721 ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag tggttcgcat cctcggtttt
      781 ctggaaggcg agcatcgttt gttcgcccag gactctagct atagttctag tggttggcta
      841 cgtatactcc ggaatattaa tagatcatgg agataattaa aatgataacc atctcgcaaa
      901 taaataagta ttttactgtt ttcgtaacag ttttgtaata aaaaaaccta taaatattcc
      961 ggattattca taccgtccca ccatcgggcg cggatctcgg tccgaaaacc atgcatcatc
     1021 atcatcatca cagcagcggc agagaaaact tgtatttcca gggcatgcag gacgcgggag
     1081 gaaacgaggg cagagcggcc gaggccgaac cccgggcgct ggagaatggc gacgcggacg
     1141 agccctcctt cagcgacccc gaggacttcg tggacgacgt gagcgaggaa gaattactgg
     1201 gagatgtact caaagatcgg ccccaggaag cagatggaat cgattcggtg attgtagtgg
     1261 acaatgtccc tcaggtggga cccgaccgac ttgagaaact caaaaatgtc atccacaaga
     1321 tcttttccaa gtttgggaaa atcacaaatg atttttatcc tgaagaggat gggaagacaa
     1381 aagggtatat tttcctggag tacgcgtccc ctgcccacgc tgtggatgct gtgaagaacg
     1441 ccgacggcta caagcttgac aagcagcaca cattccgggt caacctcttt acggattttg
     1501 acaagtatat gacgatcagt gacgagtggg atattccaga gaaacagcct ttcaaagacc
     1561 tggggaactt acgttactgg cttgaagagg cagaatgcag agatcagtac agtgtgattt
     1621 ttgagagtgg agaccgcact tccatattct ggaatgacgt aaaagaccct gtctcaattg
     1681 aagaaagagc gagatggaca gagacgtatg tgcgttggtc tcctaagggc acctacctgg
     1741 ctacctttca tcaaagaggc attgctctat gggggggaga gaaattcaag caaattcaga
     1801 gattcagcca ccaaggggtt cagcttattg acttctcacc ttgtgaaagg tacctggtga
     1861 cctttagccc cctgatggac acgcaggatg accctcaggc cataatcatc tgggacatcc
     1921 ttacggggca caagaagagg ggttttcact gtgagagctc agcccattgg cctattttta
     1981 agtggagcca tgatggcaaa ttctttgcca gaatgaccct ggatacgctt agcatctatg
     2041 aaactccttc tatgggtctt ttggacaaga agagtttgaa gatctctggg ataaaagact
     2101 tttcttggtc tcctggtggt aacataatcg ccttctgggt gcctgaagac aaagatattc
     2161 cagccagggt aaccctgatg cagctcccta ccaggcaaga gatccgagtg aggaacctgt
     2221 tcaatgtggt ggactgcaag ctccattggc agaagaacgg agactacttg tgtgtgaaag
     2281 tagataggac tccgaaaggc acccagggtg ttgtcacaaa ttttgaaatt ttccgaatga
     2341 gggagaaaca ggtacctgtg gatgtggtcg agatgaaaga aaccatcata gcctttgcct
     2401 gggaaccaaa tggaagtaag tttgctgtgc tgcacggaga ggctccgcgg atatctgtgt
     2461 ctttctacca cgtcaaaaac aacgggaaga ttgaactcat caagatgttc gacaagcagc
     2521 aggcgaacac catcttctgg agcccccaag gacagttcgt ggtgttggcg ggcctgagga
     2581 gtatgaacgg tgccttagcg tttgtggaca cttcggactg cacggtcatg aacatcgcag
     2641 agcactacat ggcttccgac gtcgaatggg atcctactgg gcgctacgtc gtcacctctg
     2701 tgtcctggtg gagccataag gtggacaacg cgtactggct gtggactttc cagggacgcc
     2761 tcctgcagaa gaacaacaag gaccgcttct gccagctgct gtggcggccc cggcctccca
     2821 cactcctgag ccaggaacag atcaagcaaa ttaaaaagga tctgaagaaa tactctaaga
     2881 tctttgaaca gaaggatcgt ttgagtcagt ccaaagcctc aaaggaattg gtggagagaa
     2941 ggcgcaccat gatggaagat ttccggaagt gccggaaaat ggcccaggag ctctatatgg
     3001 agcagaaaaa cgagcgcctg gagttgcgag gaggggtgga cactgacgag ctggacagca
     3061 acgtggacga ctgggaagag gagaccattg agttcttcgt cactgaagaa atcattcccc
     3121 tcgggaatca ggagtgatga cgaagcttgt cgagaagtac tagaggatca taatcagcca
     3181 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     3241 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     3301 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3361 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatctgat cactgatatc
     3421 gcctaggaga tccgaaccag ataagtgaaa tctagttcca aactattttg tcatttttaa
     3481 ttttcgtatt agcttacgac gctacaccca gttcccatct attttgtcac tcttccctaa
     3541 ataatcctta aaaactccat ttccacccct cccagttccc aactattttg tccgcccaca
     3601 gcggggcatt tttcttcctg ttatgttttt aatcaaacat cctgccaact ccatgtgaca
     3661 aaccgtcatc ttcggctact ttttctctgt cacagaatga aaatttttct gtcatctctt
     3721 cgttattaat gtttgtaatt gactgaatat caacgcttat ttgcagcctg aatggcgaat
     3781 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3841 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3901 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3961 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     4021 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttcttaatag
     4081 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4141 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4201 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     4261 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4321 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4381 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4441 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4501 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4561 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4621 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     4681 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4741 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     4801 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4861 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     4921 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     4981 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5041 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5101 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5161 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5221 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5281 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5341 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5401 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5461 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5521 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5581 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5641 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5701 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5761 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5821 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5881 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5941 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6001 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6061 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     6121 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     6181 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagacca gccgcgtaac
     6241 ctggcaaaat cggttacggt tgagtaataa atggatgccc tgcgtaagcg ggtgtgggcg
     6301 gacaataaag tcttaaactg aacaaaatag atctaaacta tgacaataaa gtcttaaact
     6361 agacagaata gttgtaaact gaaatcagtc cagttatgct gtgaaaaagc atactggact
     6421 tttgttatgg ctaaagcaaa ctcttcattt tctgaagtgc aaattgcccg tcgtattaaa
     6481 gaggggcgtg gccaagggca tggtaaagac tatattcgcg gcgttgtgac aatttaccga
     6541 acaactccgc ggccgggaag ccgatctcgg cttgaacgaa ttgttaggtg gcggtacttg
     6601 ggtcgatatc aaagtgcatc acttcttccc gtatgcccaa ctttgtatag agagccactg
     6661 cgggatcgtc accgtaatct gcttgcacgt agatcacata agcaccaagc gcgttggcct
     6721 catgcttgag cagattgatg agcgcggtgg caatgccctg cctccggtgc tcgccggaga
     6781 ctgcgagatc atagatatag atc
//
