LOCUS       sequence-425992-        7803 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human CBFA2T3. C-terminal 
            NanoLuc luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7803)
  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 7803)
  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_CBFA2T3:1-615
FEATURES             Location/Qualifiers
     source          1..7803
                     /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             2978..3490
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(3566..3584)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3621..3742
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3632..3651)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3686..3705
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3987..4344
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3987..4178
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4010..4030)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4195..4330
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4375..5412
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5391..5399
                     /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    5436..5484
                     /note="synthetic polyadenylation signal"
     promoter        5640..5744
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5745..5813,5814..6605)
                     /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(5963..5982)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(6763..7351)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6843..6862)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7467..7750
                     /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 gcttcaagac tgagggacag ggcagccagt tcagcctcgg gatccacctg
     1141 tggctccatg tcccagacgc accctgtgct ggagagcggc ctcctggcat ctgccggctg
     1201 ctccgcaccc cggggtccca ggaagggcgg cccagcccca gtggacagga aagctaaggc
     1261 ctcagcgatg ccggactccc cagcggaggt gaagacgcag ccccggtcca caccccccag
     1321 catgccgccc ccaccgcctg ccgcatccca gggggccaca cgccccccct ccttcacgcc
     1381 acacacactg atgaacggca gcagccactc accaacagcc atcaatggtg caccgtgcac
     1441 acccaacggc ttcagcaatg gcccggccac ctcgtccaca gcctccttgt ccacacagca
     1501 cctgccccca gcctgcgggg cccggcagct cagcaagctc aagcgcttcc tcaccacact
     1561 gcagcagttt ggcagcgaca tctccccaga gattggggag cgcgtgcgca cactggtgct
     1621 gggcctggtg aactcgacat tgacgatcga ggagtttcat tccaagcttc aggaggccac
     1681 caacttccct ctgcggccgt ttgtcattcc cttcctgaag gcaaacctgc ccttgctgca
     1741 gcgggagctc ctgcactgtg cacgcctggc caagcagacg cccgcccagt acttggccca
     1801 gcatgagcag ctcctgctgg acgccagcgc ctcctccccc atcgactcct cagagctgct
     1861 actggaagtc aacgagaacg gcaagaggag gacgcccgac aggaccaaag agaacgggtc
     1921 agaccgcgac ccgctgcacc cccagcgcta cagccccagc aacgggccac cgcagcccac
     1981 accgccgccg cactaccgcc tggaggacat agccatggcc caccacttcc gagatgccta
     2041 ccgccaccca gacccccggg agctacgaga gcgccatcgg ccgcttgtgg tgcctgggtc
     2101 ccggcaggaa gaagtgatcg accacaagct cacagagcgt gagtgggcag aagagtggaa
     2161 gcacctcaac aacctcctga actgcatcat ggacatggtg gagaagacgc ggcgctcgct
     2221 cacggtgctg cgcaggtgcc aggaggccga ccgcgaggag ctcaaccact gggcgcggcg
     2281 ctacagcgac gccgaggaca caaagaaggg ccccgctccc gccgcggccc ggccccgcag
     2341 cagctccgcc ggtcccgaag ggcctcagct agacgtgcct cgcgagttcc tgccgaggac
     2401 cctcaccggc tacgtgcctg aggacatctg gaggaaggct gaagaggccg tgaatgaggt
     2461 gaagcggcag gccatgtcgg agctgcagaa agccgtgtcg gacgcggagc gcaaagcgca
     2521 cgagctcatc accacggagc gtgccaagat ggagcgggcc ctggccgagg cgaagcggca
     2581 ggcctccgag gacgccctga cggtcatcaa ccagcaggag gactccagcg agagctgctg
     2641 gaactgcggg cggaaagcca gtgagacgtg cagcggctgc aacgcggcac gctactgcgg
     2701 gtccttctgc cagcatcggg actgggagaa gcatcaccac gtgtgtggcc agagcctgca
     2761 gggccccaca gccgtggtgg ccgacccggt gcctggaccg cccgaagccg cccacagcct
     2821 gggcccctcc ctgcctgtgg gtgctgccag ccccagcgaa gccggctctg cggggccttc
     2881 tcgccccggc tcccccagcc cacctggccc actggacacc gtgccccgct ctagatttgg
     2941 gcccaattcc tgcaggcgag ctctcggctc gagcggcgtc ttcacactcg aagatttcgt
     3001 tggggactgg cgacagacag ccggctacaa cctggaccaa gtccttgaac agggaggtgt
     3061 gtccagtttg tttcagaatc tcggggtgtc cgtaactccg atccaaagga ttgtcctgag
     3121 cggtgaaaat gggctgaaga tcgacatcca tgtcatcatc ccgtatgaag gtctgagcgg
     3181 cgaccaaatg ggccagatcg aaaaaatttt taaggtggtg taccctgtgg atgatcatca
     3241 ctttaaggtg atcctgcact atggcacact ggtaatcgac ggggttacgc cgaacatgat
     3301 cgactatttc ggacggccgt atgaaggcat cgccgtgttc gacggcaaaa agatcactgt
     3361 aacagggacc ctgtggaacg gcaacaaaat tatcgacgag cgcctgatca accccgacgg
     3421 ctccctgctg ttccgagtaa ccatcaacgg agtgaccggc tggcggctgt gcgaacgcat
     3481 tctggcgtaa ggccgcgact ctagggcatg caagctgatc cggctgctaa caaagcccga
     3541 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcg
     3601 gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     3661 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3721 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     3781 gggggagatg tgggaggttt ttttaagcaa gtaaaacctc tacaaatgtg gtaaaatcga
     3841 attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct
     3901 gtgcggtatt tcacaccgca tacgcggatc tgcgcagcac catggcctga aataacctct
     3961 gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg gaatgtgtgt
     4021 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4081 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4141 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4201 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4261 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4321 tttggaggcc taggcttttg caaaaagctc gattcttctg acactagcgc caccatgaag
     4381 aagcccgaac tcaccgctac cagcgttgaa aaatttctca tcgagaagtt cgacagtgtg
     4441 agcgacctga tgcagttgtc ggagggcgaa gagagccgag ccttcagctt cgatgtcggc
     4501 ggacgcggct atgtactgcg ggtgaatagc tgcgctgatg gcttctacaa agaccgctac
     4561 gtgtaccgcc acttcgccag cgctgcacta cccatccccg aagtgttgga catcggcgag
     4621 ttcagcgaga gcctgacata ctgcatcagt agacgcgccc aaggcgttac tctccaagac
     4681 ctccccgaaa cagagctgcc tgctgtgtta cagcctgtcg ccgaagctat ggatgctatt
     4741 gccgccgccg acctcagtca aaccagcggc ttcggcccat tcgggcccca aggcatcggc
     4801 cagtacacaa cctggcggga tttcatttgc gccattgctg atccccatgt ctaccactgg
     4861 cagaccgtga tggacgacac cgtgtccgcc agcgtagctc aagccctgga cgaactgatg
     4921 ctgtgggccg aagactgtcc cgaggtgcgc cacctcgtcc atgccgactt cggcagcaac
     4981 aacgtcctga ccgacaacgg ccgcatcacc gccgtaatcg actggtccga agctatgttc
     5041 ggggacagtc agtacgaggt ggccaacatc ttcttctggc ggccctggct ggcttgcatg
     5101 gagcagcaga ctcgctactt cgagcgccgg catcccgagc tggccggcag ccctcgtctg
     5161 cgagcctaca tgctgcgcat cggcctggat cagctctacc agagcctcgt ggacggcaac
     5221 ttcgacgatg ctgcctgggc tcaaggccgc tgcgatgcca tcgtccgcag cggggccggc
     5281 accgtcggtc gcacacaaat cgctcgccgg agcgcagccg tatggaccga cggctgcgtc
     5341 gaggtgctgg ccgacagcgg caaccgccgg cccagtacac gaccgcgcgc taaggaggta
     5401 ggtcgagttt agactctagc catcacgatg gccgcaataa aatatcttta ttttcattac
     5461 atctgtgtgt tggttttttg tgtgaatcga tagcgataag gatcctcttt gcgcttgcgt
     5521 tttcccttgt ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg
     5581 gactggcttt ctacgtaatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     5641 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     5701 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     5761 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5821 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     5881 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     5941 tgatgagcac tttcaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     6001 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     6061 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     6121 ccatgagtga taacactgcg gccaacttac ttctgacaac tatcggagga ccgaaggagc
     6181 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     6241 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     6301 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     6361 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     6421 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     6481 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6541 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6601 ggtaattcga aatgaccgac caagcgacgc ccaaccggta tcagctcact caaaggcggt
     6661 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6721 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6781 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6841 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6901 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6961 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     7021 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7081 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     7141 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     7201 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     7261 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     7321 gcagattacg cgcagaaaaa aaggatttca agaagatcct ttgatctttt ctacggggtc
     7381 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     7441 gatcttcacc tagatccttt tatagtccgg aaatacagga acgcacgctg gatggccctt
     7501 cgctgggatg gtgaaaccat gaaaaatggc agcttcagtg gattaagtgg gggtaatgtg
     7561 gcctgtaccc tctggttgca taggtattca tacggttaaa atttatcagg cgcgattgcg
     7621 gcagtttttc gggtggtttg ttgccatttt tacctgtctg ctgccgtgat cgcgctgaac
     7681 gcgttttagc ggtgcgtaca attaagggat tatggtaaat ccacttactg tctgccctcg
     7741 tagccatcga gataaaccgc agtactccgg ccacgatgcg tccggcgtag aggatcgaga
     7801 tct
//
