LOCUS       sequence-426246-        6014 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect expression of human EIF2A fragment M1 to T427. N-terminal 6X 
            His tag with TEV cleavage..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6014)
  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 6014)
  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_EIF2A:1-427
FEATURES             Location/Qualifiers
     source          1..6014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    875..1009
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(912..931)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     966..985
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1038..1203
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      1387..1841
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1474..1493)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1683..1704
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1867..1971
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1972..2040,2041..2832)
                     /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(2190..2209)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3003..3591
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3492..3511
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3745..3762
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  3896..4120
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4187..4720)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4890..4895,4896..4912,4913..4918))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        5270..5361
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     5286..5303
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             5420..5437
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             5450..5470
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 acaaggtggc tgtctatgtt ccaggaagta aaggtgcacc ttcatttgtt agattatatc
       61 agtaccccaa ctttgctgga cctcatgcag ctttagctaa taaaagtttc tttaaggcag
      121 ataaagttac aatgctgtgg aataaaaaag ctactgctgt gttggtaata gctagcacag
      181 atgttgacaa gacaggagct tcctactatg gagaacaaac tctacactac attgcaacaa
      241 atggagaaag tgctgtagtg caattaccaa aaaatggccc catttatgat gtagtttgga
      301 attctagttc tactgagttt tgtgctgtat atggttttat gcctgccaaa gcgacaattt
      361 tcaacttgaa atgtgatcct gtatttgact ttggaactgg tcctcgtaat gcagcctact
      421 atagccctca tggacatata ttagtattag ctggatttgg aaatctgagg ggacaaatgg
      481 aagtgtggga tgtgaaaaac tacaaactta tttctaaacc ggtggcttct gattctacat
      541 attttgcttg gtgcccggat ggtgagcata ttttaacagc tacatgtgct cccaggttac
      601 gggttaataa tggatacaaa atttggcatt atactggctc tatcttgcac aagtatgatg
      661 tgccatcaaa tgcagaatta tggcaggttt cttggcagcc atttttggat ggaatatttc
      721 cagcaaaaac aataacttga tgacgaagct tgtcgagaag tactagagga tcataatcag
      781 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
      841 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
      901 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
      961 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct gatcactgat
     1021 atcgcctagg agatccgaac cagataagtg aaatctagtt ccaaactatt ttgtcatttt
     1081 taattttcgt attagcttac gacgctacac ccagttccca tctattttgt cactcttccc
     1141 taaataatcc ttaaaaactc catttccacc cctcccagtt cccaactatt ttgtccgccc
     1201 acagcggggc atttttcttc ctgttatgtt tttaatcaaa catcctgcca actccatgtg
     1261 acaaaccgtc atcttcggct actttttctc tgtcacagaa tgaaaatttt tctgtcatct
     1321 cttcgttatt aatgtttgta attgactgaa tatcaacgct tatttgcagc ctgaatggcg
     1381 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     1441 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     1501 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     1561 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     1621 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttaa
     1681 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     1741 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     1801 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     1861 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     1921 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     1981 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     2041 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2101 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2161 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     2221 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2281 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2341 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2401 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2461 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     2521 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2581 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2641 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     2701 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     2761 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     2821 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     2881 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     2941 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     3001 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3061 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3121 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     3181 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3241 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3301 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3361 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3421 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3481 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3541 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3601 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3661 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3721 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     3781 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcataga ccagccgcgt
     3841 aacctggcaa aatcggttac ggttgagtaa taaatggatg ccctgcgtaa gcgggtgtgg
     3901 gcggacaata aagtcttaaa ctgaacaaaa tagatctaaa ctatgacaat aaagtcttaa
     3961 actagacaga atagttgtaa actgaaatca gtccagttat gctgtgaaaa agcatactgg
     4021 acttttgtta tggctaaagc aaactcttca ttttctgaag tgcaaattgc ccgtcgtatt
     4081 aaagaggggc gtggccaagg gcatggtaaa gactatattc gcggcgttgt gacaatttac
     4141 cgaacaactc cgcggccggg aagccgatct cggcttgaac gaattgttag gtggcggtac
     4201 ttgggtcgat atcaaagtgc atcacttctt cccgtatgcc caactttgta tagagagcca
     4261 ctgcgggatc gtcaccgtaa tctgcttgca cgtagatcac ataagcacca agcgcgttgg
     4321 cctcatgctt gagcagattg atgagcgcgg tggcaatgcc ctgcctccgg tgctcgccgg
     4381 agactgcgag atcatagata tagatctcac tacgcggctg ctcaaacttg ggcagaacgt
     4441 aagccgcgag agcgccaaca accgcttctt ggtcgaaggc agcaagcgcg atgaatgtct
     4501 tactacggag caagttcccg aggtaatcgg agtccggctg atgttgggag taggtggcta
     4561 cgtctccgaa ctcacgaccg aaaagatcaa gagcagcccg catggatttg acttggtcag
     4621 ggccgagcct acatgtgcga atgatgccca tacttgagcc acctaacttt gttttagggc
     4681 gactgccctg ctgcgtaaca tcgttgctgc tccataacat caaacatcga cccacggcgt
     4741 aacgcgcttg ctgcttggat gcccgaggca tagactgtac aaaaaaacag tcataacaag
     4801 ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt
     4861 tgcgtgagcg catacgctac ttgcattaca gtttacgaac cgaacaggct tatgtcaact
     4921 gggttcgtgc cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg ggcagcagcg
     4981 aagtcgaggc atttctgtcc tggctggcga acgagcgcaa ggtttcggtc tccacgcatc
     5041 gtcaggcatt ggcggccttg ctgttcttct acggcaaggt gctgtgcacg gatctgccct
     5101 ggcttcagga gatcggaaga cctcggccgt cgcggcgctt gccggtggtg ctgaccccgg
     5161 atgaagtggt tcgcatcctc ggttttctgg aaggcgagca tcgtttgttc gcccaggact
     5221 ctagctatag ttctagtggt tggctacgta tactccggaa tattaataga tcatggagat
     5281 aattaaaatg ataaccatct cgcaaataaa taagtatttt actgttttcg taacagtttt
     5341 gtaataaaaa aacctataaa tattccggat tattcatacc gtcccaccat cgggcgcgga
     5401 tctcggtccg aaaaccatgc atcatcatca tcatcacagc agcggcagag aaaacttgta
     5461 tttccagggc atggcgccgt ccacgccgct cttgacagtc cgaggatcag aaggactgta
     5521 catggtgaat ggaccaccac attttacaga aagcacagtg tttccaaggg aatctgggaa
     5581 gaattgcaaa gtctgtatct ttagtaagga tgggaccttg tttgcctggg gcaatggaga
     5641 aaaagtaaat attatcagtg tcactaacaa gggactactg cactccttcg acctcctgaa
     5701 ggcagtttgc cttgaattct cacccaaaaa tactgtcctg gcaacgtggc agccttacac
     5761 tacttctaaa gatggcacag ctgggatacc caacctacaa ctttatgatg tgaaaactgg
     5821 gacatgtttg aaatctttca tccagaaaaa aatgcaaaat tggtgtccat cctggtcaga
     5881 agatgaaact ctttgtgccc gcaatgttaa caatgaagtt cacttctttg aaaacaacaa
     5941 ttttaacaca attgcaaata aattgcattt gcaaaaaatt aatgactttg tattatcacc
     6001 tggaccccaa ccat
//
