LOCUS       sequence-423317-        6900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human MAGEA3. C-terminal NanoLuc
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6900)
  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 6900)
  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_MAGEA3:1-314
FEATURES             Location/Qualifiers
     source          1..6900
                     /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             2075..2587
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2663..2681)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2718..2839
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2729..2748)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2783..2802
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3084..3441
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3084..3275
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3107..3127)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3292..3427
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3472..4509
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4488..4496
                     /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    4533..4581
                     /note="synthetic polyadenylation signal"
     promoter        4737..4841
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4842..4910,4911..5702)
                     /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(5060..5079)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5860..6448)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5940..5959)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6564..6847
                     /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 caccatgcct cttgagcaga ggagtcagca ctgcaagcct gaagaaggcc ttgaggcccg
     1141 aggagaggcc ctgggcctgg tgggtgcgca ggctcctgct actgaggagc aggaggctgc
     1201 ctcctcctct tctactctag ttgaagtcac cctgggggag gtgcctgctg ccgagtcacc
     1261 agatcctccc cagagtcctc agggagcctc cagcctcccc actaccatga actaccctct
     1321 ctggagccaa tcctatgagg actccagcaa ccaagaagag gaggggccaa gcaccttccc
     1381 tgacctggag tccgagttcc aagcagcact cagtaggaag gtggccgagt tggttcattt
     1441 tctgctcctc aagtatcgag ccagggagcc ggtcacaaag gcagaaatgc tggggagtgt
     1501 cgtcggaaat tggcagtatt tctttcctgt gatcttcagc aaagcttcca gttccttgca
     1561 gctggtcttt ggcatcgagc tgatggaagt ggaccccatc ggccacttgt acatctttgc
     1621 cacctgcctg ggcctctcct acgatggcct gctgggtgac aatcagatca tgcccaaggc
     1681 aggcctcctg ataatcgtcc tggccataat cgcaagagag ggcgactgtg cccctgagga
     1741 gaaaatctgg gaggagctga gtgtgttaga ggtgtttgag gggagggaag acagtatctt
     1801 gggggatccc aagaagctgc tcacccaaca tttcgtgcag gaaaactacc tggagtaccg
     1861 gcaggtcccc ggcagtgatc ctgcatgtta tgaattcctg tggggtccaa gggccctcgt
     1921 tgaaaccagc tatgtgaaag tcctgcacca tatggtaaag atcagtggag gacctcacat
     1981 ttcctaccca cccctgcatg agtgggtttt gagagagggg gaagagtcta gatttgggcc
     2041 caattcctgc aggcgagctc tcggctcgag cggcgtcttc acactcgaag atttcgttgg
     2101 ggactggcga cagacagccg gctacaacct ggaccaagtc cttgaacagg gaggtgtgtc
     2161 cagtttgttt cagaatctcg gggtgtccgt aactccgatc caaaggattg tcctgagcgg
     2221 tgaaaatggg ctgaagatcg acatccatgt catcatcccg tatgaaggtc tgagcggcga
     2281 ccaaatgggc cagatcgaaa aaatttttaa ggtggtgtac cctgtggatg atcatcactt
     2341 taaggtgatc ctgcactatg gcacactggt aatcgacggg gttacgccga acatgatcga
     2401 ctatttcgga cggccgtatg aaggcatcgc cgtgttcgac ggcaaaaaga tcactgtaac
     2461 agggaccctg tggaacggca acaaaattat cgacgagcgc ctgatcaacc ccgacggctc
     2521 cctgctgttc cgagtaacca tcaacggagt gaccggctgg cggctgtgcg aacgcattct
     2581 ggcgtaaggc cgcgactcta gggcatgcaa gctgatccgg ctgctaacaa agcccgaaag
     2641 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcggcc
     2701 gcttcgagca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
     2761 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
     2821 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     2881 ggagatgtgg gaggtttttt taagcaagta aaacctctac aaatgtggta aaatcgaatt
     2941 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     3001 cggtatttca caccgcatac gcggatctgc gcagcaccat ggcctgaaat aacctctgaa
     3061 agaggaactt ggttaggtac cttctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3121 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3181 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3241 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3301 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3361 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3421 ggaggcctag gcttttgcaa aaagctcgat tcttctgaca ctagcgccac catgaagaag
     3481 cccgaactca ccgctaccag cgttgaaaaa tttctcatcg agaagttcga cagtgtgagc
     3541 gacctgatgc agttgtcgga gggcgaagag agccgagcct tcagcttcga tgtcggcgga
     3601 cgcggctatg tactgcgggt gaatagctgc gctgatggct tctacaaaga ccgctacgtg
     3661 taccgccact tcgccagcgc tgcactaccc atccccgaag tgttggacat cggcgagttc
     3721 agcgagagcc tgacatactg catcagtaga cgcgcccaag gcgttactct ccaagacctc
     3781 cccgaaacag agctgcctgc tgtgttacag cctgtcgccg aagctatgga tgctattgcc
     3841 gccgccgacc tcagtcaaac cagcggcttc ggcccattcg ggccccaagg catcggccag
     3901 tacacaacct ggcgggattt catttgcgcc attgctgatc cccatgtcta ccactggcag
     3961 accgtgatgg acgacaccgt gtccgccagc gtagctcaag ccctggacga actgatgctg
     4021 tgggccgaag actgtcccga ggtgcgccac ctcgtccatg ccgacttcgg cagcaacaac
     4081 gtcctgaccg acaacggccg catcaccgcc gtaatcgact ggtccgaagc tatgttcggg
     4141 gacagtcagt acgaggtggc caacatcttc ttctggcggc cctggctggc ttgcatggag
     4201 cagcagactc gctacttcga gcgccggcat cccgagctgg ccggcagccc tcgtctgcga
     4261 gcctacatgc tgcgcatcgg cctggatcag ctctaccaga gcctcgtgga cggcaacttc
     4321 gacgatgctg cctgggctca aggccgctgc gatgccatcg tccgcagcgg ggccggcacc
     4381 gtcggtcgca cacaaatcgc tcgccggagc gcagccgtat ggaccgacgg ctgcgtcgag
     4441 gtgctggccg acagcggcaa ccgccggccc agtacacgac cgcgcgctaa ggaggtaggt
     4501 cgagtttaga ctctagccat cacgatggcc gcaataaaat atctttattt tcattacatc
     4561 tgtgtgttgg ttttttgtgt gaatcgatag cgataaggat cctctttgcg cttgcgtttt
     4621 cccttgtcca gatagcccag tagctgacat tcatccgggg tcagcaccgt ttctgcggac
     4681 tggctttcta cgtaatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4741 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4801 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4861 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4921 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4981 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5041 tgagcacttt caaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5101 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5161 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5221 tgagtgataa cactgcggcc aacttacttc tgacaactat cggaggaccg aaggagctaa
     5281 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5341 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5401 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5461 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5521 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5581 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5641 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5701 aattcgaaat gaccgaccaa gcgacgccca accggtatca gctcactcaa aggcggtaat
     5761 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5821 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5881 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5941 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6001 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6061 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6121 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6181 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6241 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6301 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6361 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6421 gattacgcgc agaaaaaaag gatttcaaga agatcctttg atcttttcta cggggtctga
     6481 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6541 cttcacctag atccttttat agtccggaaa tacaggaacg cacgctggat ggcccttcgc
     6601 tgggatggtg aaaccatgaa aaatggcagc ttcagtggat taagtggggg taatgtggcc
     6661 tgtaccctct ggttgcatag gtattcatac ggttaaaatt tatcaggcgc gattgcggca
     6721 gtttttcggg tggtttgttg ccatttttac ctgtctgctg ccgtgatcgc gctgaacgcg
     6781 ttttagcggt gcgtacaatt aagggattat ggtaaatcca cttactgtct gccctcgtag
     6841 ccatcgagat aaaccgcagt actccggcca cgatgcgtcc ggcgtagagg atcgagatct
//
