LOCUS       sequence-426005-        7011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human EIF2B2. C-terminal NanoLuc
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7011)
  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 7011)
  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: pNLF1-C_EIF2B2:1-351
FEATURES             Location/Qualifiers
     source          1..7011
                     /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             2186..2698
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2774..2792)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2829..2950
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2840..2859)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2894..2913
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3195..3552
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3195..3386
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3218..3238)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3403..3538
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3583..4620
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4599..4607
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    4644..4692
                     /note="synthetic polyadenylation signal"
     promoter        4848..4952
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4953..5021,5022..5813)
                     /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(5171..5190)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5971..6559)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6051..6070)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6675..6958
                     /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 ccgaattcgc
     1081 caccatgccg ggatccgcag cgaagggctc ggagttgtca gagaggatcg agagcttcgt
     1141 ggagaccctg aagcggggtg gtgggccgcg cagctccgag gaaatggctc gggagaccct
     1201 agggttgctg cgccagatca tcacggacca ccgctggagc aacgcggggg agctgatgga
     1261 gctgatccgc agagagggca ggaggatgac ggccgctcag ccctccgaga ccaccgtggg
     1321 caacatggtg cggagagtgc tcaagattat ccgggaggag tatggcagac tccatggacg
     1381 cagcgacgag agtgatcagc aggagtccct gcacaaactg ttgacatccg gaggcctaaa
     1441 cgaggatttc agcttccatt atgcccaact ccagtccaac atcattgagg cgattaatga
     1501 gctgctagtg gagctggaag ggacaatgga gaacattgca gcccaggctc tggagcacat
     1561 tcactccaat gaggtgatca tgaccattgg cttctcccga acagtagagg ccttcctcaa
     1621 agaggctgcc cgaaagagga aattccatgt cattgtagca gagtgtgctc ctttctgcca
     1681 gggtcatgaa atggctgtga atttgtccaa agcaggtatt gagacaactg tcatgactga
     1741 tgctgccatt tttgccgtta tgtcaagagt caacaaggtg atcattggca cgaagaccat
     1801 cctggccaat ggggccctga gagctgtgac aggaactcac actctggcac tggcagcaaa
     1861 acaccattcc accccactca tcgtctgtgc acctatgttc aaactttctc cacagttccc
     1921 caatgaagaa gactcatttc ataagtttgt ggctcctgaa gaagtcctgc cattcacaga
     1981 aggggacatt ctggagaagg tcagcgtgca ttgccctgtg tttgactacg ttcccccaga
     2041 gctcattacc ctctttatct ccaacattgg tgggaatgca ccttcctaca tctaccgcct
     2101 gatgagtgaa ctctaccatc ctgatgatca tgttttatct agatttgggc ccaattcctg
     2161 caggcgagct ctcggctcga gcggcgtctt cacactcgaa gatttcgttg gggactggcg
     2221 acagacagcc ggctacaacc tggaccaagt ccttgaacag ggaggtgtgt ccagtttgtt
     2281 tcagaatctc ggggtgtccg taactccgat ccaaaggatt gtcctgagcg gtgaaaatgg
     2341 gctgaagatc gacatccatg tcatcatccc gtatgaaggt ctgagcggcg accaaatggg
     2401 ccagatcgaa aaaattttta aggtggtgta ccctgtggat gatcatcact ttaaggtgat
     2461 cctgcactat ggcacactgg taatcgacgg ggttacgccg aacatgatcg actatttcgg
     2521 acggccgtat gaaggcatcg ccgtgttcga cggcaaaaag atcactgtaa cagggaccct
     2581 gtggaacggc aacaaaatta tcgacgagcg cctgatcaac cccgacggct ccctgctgtt
     2641 ccgagtaacc atcaacggag tgaccggctg gcggctgtgc gaacgcattc tggcgtaagg
     2701 ccgcgactct agggcatgca agctgatccg gctgctaaca aagcccgaaa ggaagctgag
     2761 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcggc cgcttcgagc
     2821 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     2881 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     2941 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg
     3001 ggaggttttt ttaagcaagt aaaacctcta caaatgtggt aaaatcgaat tttaacaaaa
     3061 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3121 acaccgcata cgcggatctg cgcagcacca tggcctgaaa taacctctga aagaggaact
     3181 tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     3241 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3301 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3361 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3421 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3481 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3541 ggcttttgca aaaagctcga ttcttctgac actagcgcca ccatgaagaa gcccgaactc
     3601 accgctacca gcgttgaaaa atttctcatc gagaagttcg acagtgtgag cgacctgatg
     3661 cagttgtcgg agggcgaaga gagccgagcc ttcagcttcg atgtcggcgg acgcggctat
     3721 gtactgcggg tgaatagctg cgctgatggc ttctacaaag accgctacgt gtaccgccac
     3781 ttcgccagcg ctgcactacc catccccgaa gtgttggaca tcggcgagtt cagcgagagc
     3841 ctgacatact gcatcagtag acgcgcccaa ggcgttactc tccaagacct ccccgaaaca
     3901 gagctgcctg ctgtgttaca gcctgtcgcc gaagctatgg atgctattgc cgccgccgac
     3961 ctcagtcaaa ccagcggctt cggcccattc gggccccaag gcatcggcca gtacacaacc
     4021 tggcgggatt tcatttgcgc cattgctgat ccccatgtct accactggca gaccgtgatg
     4081 gacgacaccg tgtccgccag cgtagctcaa gccctggacg aactgatgct gtgggccgaa
     4141 gactgtcccg aggtgcgcca cctcgtccat gccgacttcg gcagcaacaa cgtcctgacc
     4201 gacaacggcc gcatcaccgc cgtaatcgac tggtccgaag ctatgttcgg ggacagtcag
     4261 tacgaggtgg ccaacatctt cttctggcgg ccctggctgg cttgcatgga gcagcagact
     4321 cgctacttcg agcgccggca tcccgagctg gccggcagcc ctcgtctgcg agcctacatg
     4381 ctgcgcatcg gcctggatca gctctaccag agcctcgtgg acggcaactt cgacgatgct
     4441 gcctgggctc aaggccgctg cgatgccatc gtccgcagcg gggccggcac cgtcggtcgc
     4501 acacaaatcg ctcgccggag cgcagccgta tggaccgacg gctgcgtcga ggtgctggcc
     4561 gacagcggca accgccggcc cagtacacga ccgcgcgcta aggaggtagg tcgagtttag
     4621 actctagcca tcacgatggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg
     4681 gttttttgtg tgaatcgata gcgataagga tcctctttgc gcttgcgttt tcccttgtcc
     4741 agatagccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct
     4801 acgtaatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4861 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4921 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     4981 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5041 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5101 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5161 tcaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5221 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5281 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5341 acactgcggc caacttactt ctgacaacta tcggaggacc gaaggagcta accgcttttt
     5401 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5461 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5521 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5581 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5641 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     5701 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     5761 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taattcgaaa
     5821 tgaccgacca agcgacgccc aaccggtatc agctcactca aaggcggtaa tacggttatc
     5881 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5941 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6001 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6061 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6121 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6181 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6241 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6301 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6361 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     6421 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6481 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6541 cagaaaaaaa ggatttcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6601 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6661 gatcctttta tagtccggaa atacaggaac gcacgctgga tggcccttcg ctgggatggt
     6721 gaaaccatga aaaatggcag cttcagtgga ttaagtgggg gtaatgtggc ctgtaccctc
     6781 tggttgcata ggtattcata cggttaaaat ttatcaggcg cgattgcggc agtttttcgg
     6841 gtggtttgtt gccattttta cctgtctgct gccgtgatcg cgctgaacgc gttttagcgg
     6901 tgcgtacaat taagggatta tggtaaatcc acttactgtc tgccctcgta gccatcgaga
     6961 taaaccgcag tactccggcc acgatgcgtc cggcgtagag gatcgagatc t
//
