LOCUS       sequence-425971-        6897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human UNG. C-terminal NanoLuc 
            luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6897)
  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 6897)
  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_UNG:1-313
FEATURES             Location/Qualifiers
     source          1..6897
                     /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             2072..2584
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2660..2678)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2715..2836
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2726..2745)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2780..2799
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3081..3438
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3081..3272
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3104..3124)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3289..3424
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3469..4506
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             4485..4493
                     /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    4530..4578
                     /note="synthetic polyadenylation signal"
     promoter        4734..4838
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4839..4907,4908..5699)
                     /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(5057..5076)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5857..6445)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5937..5956)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6561..6844
                     /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 caccatgatc ggccagaaga cgctctactc ctttttctcc cccagccccg ccaggaagcg
     1141 acacgccccc agccccgagc cggccgtcca ggggaccggc gtggctgggg tgcctgagga
     1201 aagcggagat gcggcggcca tcccagccaa gaaggccccg gctgggcagg aggagcctgg
     1261 gacgccgccc tcctcgccgc tgagtgccga gcagttggac cggatccaga ggaacaaggc
     1321 cgcggccctg ctcagactcg cggcccgcaa cgtgcccgtg ggctttggag agagctggaa
     1381 gaagcacctc agcggggagt tcgggaaacc gtattttatc aagctaatgg gatttgttgc
     1441 agaagaaaga aagcattaca ctgtttatcc acccccacac caagtcttca cctggaccca
     1501 gatgtgtgac ataaaagatg tgaaggttgt catcctggga caggatccat atcatggacc
     1561 taatcaagct cacgggctct gctttagtgt tcaaaggcct gttccgcctc cgcccagttt
     1621 ggagaacatt tataaagagt tgtctacaga catagaggat tttgttcatc ctggccatgg
     1681 agatttatct gggtgggcca agcaaggtgt tctccttctc aacgctgtcc tcacggttcg
     1741 tgcccatcaa gccaactctc ataaggagcg aggctgggag cagttcactg atgcagttgt
     1801 gtcctggcta aatcagaact cgaatggcct tgttttcttg ctctggggct cttatgctca
     1861 gaagaagggc agtgccattg ataggaagcg gcaccatgta ctacagacgg ctcatccctc
     1921 ccctttgtca gtgtatagag ggttctttgg atgtagacac ttttcaaaga ccaatgagct
     1981 gctgcagaag tctggcaaga agcccattga ctggaaggag ctgtctagat ttgggcccaa
     2041 ttcctgcagg cgagctctcg gctcgagcgg cgtcttcaca ctcgaagatt tcgttgggga
     2101 ctggcgacag acagccggct acaacctgga ccaagtcctt gaacagggag gtgtgtccag
     2161 tttgtttcag aatctcgggg tgtccgtaac tccgatccaa aggattgtcc tgagcggtga
     2221 aaatgggctg aagatcgaca tccatgtcat catcccgtat gaaggtctga gcggcgacca
     2281 aatgggccag atcgaaaaaa tttttaaggt ggtgtaccct gtggatgatc atcactttaa
     2341 ggtgatcctg cactatggca cactggtaat cgacggggtt acgccgaaca tgatcgacta
     2401 tttcggacgg ccgtatgaag gcatcgccgt gttcgacggc aaaaagatca ctgtaacagg
     2461 gaccctgtgg aacggcaaca aaattatcga cgagcgcctg atcaaccccg acggctccct
     2521 gctgttccga gtaaccatca acggagtgac cggctggcgg ctgtgcgaac gcattctggc
     2581 gtaaggccgc gactctaggg catgcaagct gatccggctg ctaacaaagc ccgaaaggaa
     2641 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcggccgct
     2701 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     2761 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     2821 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     2881 gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgaatttta
     2941 acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg
     3001 tatttcacac cgcatacgcg gatctgcgca gcaccatggc ctgaaataac ctctgaaaga
     3061 ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta
     3121 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3181 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3241 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3301 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3361 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3421 ggcctaggct tttgcaaaaa gctcgattct tctgacacta gcgccaccat gaagaagccc
     3481 gaactcaccg ctaccagcgt tgaaaaattt ctcatcgaga agttcgacag tgtgagcgac
     3541 ctgatgcagt tgtcggaggg cgaagagagc cgagccttca gcttcgatgt cggcggacgc
     3601 ggctatgtac tgcgggtgaa tagctgcgct gatggcttct acaaagaccg ctacgtgtac
     3661 cgccacttcg ccagcgctgc actacccatc cccgaagtgt tggacatcgg cgagttcagc
     3721 gagagcctga catactgcat cagtagacgc gcccaaggcg ttactctcca agacctcccc
     3781 gaaacagagc tgcctgctgt gttacagcct gtcgccgaag ctatggatgc tattgccgcc
     3841 gccgacctca gtcaaaccag cggcttcggc ccattcgggc cccaaggcat cggccagtac
     3901 acaacctggc gggatttcat ttgcgccatt gctgatcccc atgtctacca ctggcagacc
     3961 gtgatggacg acaccgtgtc cgccagcgta gctcaagccc tggacgaact gatgctgtgg
     4021 gccgaagact gtcccgaggt gcgccacctc gtccatgccg acttcggcag caacaacgtc
     4081 ctgaccgaca acggccgcat caccgccgta atcgactggt ccgaagctat gttcggggac
     4141 agtcagtacg aggtggccaa catcttcttc tggcggccct ggctggcttg catggagcag
     4201 cagactcgct acttcgagcg ccggcatccc gagctggccg gcagccctcg tctgcgagcc
     4261 tacatgctgc gcatcggcct ggatcagctc taccagagcc tcgtggacgg caacttcgac
     4321 gatgctgcct gggctcaagg ccgctgcgat gccatcgtcc gcagcggggc cggcaccgtc
     4381 ggtcgcacac aaatcgctcg ccggagcgca gccgtatgga ccgacggctg cgtcgaggtg
     4441 ctggccgaca gcggcaaccg ccggcccagt acacgaccgc gcgctaagga ggtaggtcga
     4501 gtttagactc tagccatcac gatggccgca ataaaatatc tttattttca ttacatctgt
     4561 gtgttggttt tttgtgtgaa tcgatagcga taaggatcct ctttgcgctt gcgttttccc
     4621 ttgtccagat agcccagtag ctgacattca tccggggtca gcaccgtttc tgcggactgg
     4681 ctttctacgt aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4741 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4801 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4861 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4921 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4981 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5041 gcactttcaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5101 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5161 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5221 gtgataacac tgcggccaac ttacttctga caactatcgg aggaccgaag gagctaaccg
     5281 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5341 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5401 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5461 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5521 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5581 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5641 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaat
     5701 tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct cactcaaagg cggtaatacg
     5761 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5821 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5881 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5941 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6001 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6061 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6121 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     6181 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     6241 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     6301 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     6361 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     6421 tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     6481 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6541 cacctagatc cttttatagt ccggaaatac aggaacgcac gctggatggc ccttcgctgg
     6601 gatggtgaaa ccatgaaaaa tggcagcttc agtggattaa gtgggggtaa tgtggcctgt
     6661 accctctggt tgcataggta ttcatacggt taaaatttat caggcgcgat tgcggcagtt
     6721 tttcgggtgg tttgttgcca tttttacctg tctgctgccg tgatcgcgct gaacgcgttt
     6781 tagcggtgcg tacaattaag ggattatggt aaatccactt actgtctgcc ctcgtagcca
     6841 tcgagataaa ccgcagtact ccggccacga tgcgtccggc gtagaggatc gagatct
//
