LOCUS       sequence-425968-        8301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of full-length human PML isoform 13. C-terminal
            NanoLuc luciferase tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8301)
  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 8301)
  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_PML:1-781
FEATURES             Location/Qualifiers
     source          1..8301
                     /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"
     regulatory      1079..1088
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3476..3988
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     polyA_signal    4119..4240
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4130..4149)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4184..4203
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4485..4842
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4485..4676
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4508..4528)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4693..4828
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4873..5910
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             5889..5897
                     /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    5934..5982
                     /note="synthetic polyadenylation signal"
     promoter        6138..6242
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6243..6311,6312..7103)
                     /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(6461..6480)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(7261..7849)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7341..7360)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    7965..8248
                     /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 caccatggag cctgcacccg cccgatctcc gaggccccag caggaccccg cccggcccca
     1141 ggagcccacc atgcctcccc ccgagacccc ctctgaaggc cgccagccca gccccagccc
     1201 cagccctaca gagcgagccc ccgcttcgga ggaggagttc cagtttctgc gctgccagca
     1261 atgccaggcg gaagccaagt gcccgaagct gctgccttgt ctgcacacgc tgtgctcagg
     1321 atgcctggag gcgtcgggca tgcagtgccc catctgccag gcgccctggc ccctaggtgc
     1381 agacacaccc gccctggata acgtcttttt cgagagtctg cagcggcgcc tgtcggtgta
     1441 ccggcagatt gtggatgcgc aggctgtgtg cacccgctgc aaagagtcgg ccgacttctg
     1501 gtgctttgag tgcgagcagc tcctctgcgc caagtgcttc gaggcacacc agtggttcct
     1561 caagcacgag gcccggcccc tagcagagct gcgcaaccag tcggtgcgtg agttcctgga
     1621 cggcacccgc aagaccaaca acatcttctg ctccaacccc aaccaccgca cccctacgct
     1681 gaccagcatc tactgccgag gatgttccaa gccgctgtgc tgctcgtgcg cgctccttga
     1741 cagcagccac agtgatctca agtgcgacat cagcgcagag atccagcagc gacaggagga
     1801 gctggacgcc atgacgcagg cgctgcagga gcaggatagt gcctttggcg cggttcacgc
     1861 gcagatgcac gcggctgtcg gccagctggg ccgcgcgcgt gccgagaccg aggagctgat
     1921 ccgcgagcgc gtgcgccagg tggtagctca cgtgcgggct caggagcgcg agctgctgga
     1981 ggctgtggac gcgcggtacc agcgcgacta cgaggagatg gccagtcggc tgggccgcct
     2041 ggatgctgtg ctgcagcgca tccgcacggg cagcgcgctg gtgcagagga tgaagtgcta
     2101 cgcctcggac caggaggtgc tggacatgca cggtttcctg cgccaggcgc tctgccgcct
     2161 gcgccaggag gagccccaga gcctgcaagc tgccgtgcgc accgatggct tcgacgagtt
     2221 caaggtgcgc ctgcaggacc tcagctcttg catcacccag gggaaagatg cagctgtatc
     2281 caagaaagcc agcccagagg ctgccagcac tcccagggac cctattgacg ttgacctgga
     2341 tgtctccaat acaacgacag cccagaagag gaagtgcagc cagacccagt gccccaggaa
     2401 ggtcatcaag atggagtctg aggaggggaa ggaggcaagg ttggctcgga gctccccgga
     2461 gcagcccagg cccagcacct ccaaggcagt ctcaccaccc cacctggatg gaccgcctag
     2521 ccccaggagc cccgtcatag gaagtgaggt cttcctgccc aacagcaacc acgtggccag
     2581 tggcgccggg gaggcagagg aacgcgttgt ggtgatcagc agctcggaag actcagatgc
     2641 cgaaaactcg tgcatggagc ccatggagac cgccgagcca cagtcctcgc cagcccactc
     2701 ctcgccagcc cactcctcgc cagcccactc ctcgccagtc cagtctctgc tgagagcaca
     2761 aggagcctcc agcctgccct gtggcacata ccacccccca gcttggcctc cccaccagcc
     2821 cgctgagcag gctgccaccc ccgatgctga gcctcacagc gagcctcctg atcaccagga
     2881 gcgccctgcc gtccaccgtg ggatccgcta cctgttgtac agagcacaga gagccatccg
     2941 ccttcgccat gccctccgct tgcaccctca attgcatcgg gcccctattc ggacttggtc
     3001 tccccatgtg gtccaagcca gcactcctgc catcacaggg cccctcaacc atcctgccaa
     3061 tgcccaggaa catcctgccc agctgcaaag gggcatcagc ccaccccacc ggatacgagg
     3121 ggctgtgcga tcccgcagcc gctccctccg gggctcctcc catttatccc agtggctcaa
     3181 caactttttt gccctcccct tctcctccat ggcttcccag cttgacatgt cttccgtggt
     3241 gggggcaggg gaaggcagag cccagactct tggagcagtt gttccccctg gggactctgt
     3301 cagaggctcc atggaggcct ctcaagtcca agtgcctctg gaagcctctc caattacatt
     3361 cccaccaccc tgtgccccag aaaggccccc catcagccca gtcccaggcg cccgtcaagc
     3421 aggcctctct agatttgggc ccaattcctg caggcgagct ctcggctcga gcggcgtctt
     3481 cacactcgaa gatttcgttg gggactggcg acagacagcc ggctacaacc tggaccaagt
     3541 ccttgaacag ggaggtgtgt ccagtttgtt tcagaatctc ggggtgtccg taactccgat
     3601 ccaaaggatt gtcctgagcg gtgaaaatgg gctgaagatc gacatccatg tcatcatccc
     3661 gtatgaaggt ctgagcggcg accaaatggg ccagatcgaa aaaattttta aggtggtgta
     3721 ccctgtggat gatcatcact ttaaggtgat cctgcactat ggcacactgg taatcgacgg
     3781 ggttacgccg aacatgatcg actatttcgg acggccgtat gaaggcatcg ccgtgttcga
     3841 cggcaaaaag atcactgtaa cagggaccct gtggaacggc aacaaaatta tcgacgagcg
     3901 cctgatcaac cccgacggct ccctgctgtt ccgagtaacc atcaacggag tgaccggctg
     3961 gcggctgtgc gaacgcattc tggcgtaagg ccgcgactct agggcatgca agctgatccg
     4021 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     4081 gcataacccc ttggggcggc cgcttcgagc agacatgata agatacattg atgagtttgg
     4141 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     4201 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     4261 ttttatgttt caggttcagg gggagatgtg ggaggttttt ttaagcaagt aaaacctcta
     4321 caaatgtggt aaaatcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta
     4381 ttttctcctt acgcatctgt gcggtatttc acaccgcata cgcggatctg cgcagcacca
     4441 tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg cggaaagaac
     4501 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4561 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4621 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4681 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4741 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4801 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctcga ttcttctgac
     4861 actagcgcca ccatgaagaa gcccgaactc accgctacca gcgttgaaaa atttctcatc
     4921 gagaagttcg acagtgtgag cgacctgatg cagttgtcgg agggcgaaga gagccgagcc
     4981 ttcagcttcg atgtcggcgg acgcggctat gtactgcggg tgaatagctg cgctgatggc
     5041 ttctacaaag accgctacgt gtaccgccac ttcgccagcg ctgcactacc catccccgaa
     5101 gtgttggaca tcggcgagtt cagcgagagc ctgacatact gcatcagtag acgcgcccaa
     5161 ggcgttactc tccaagacct ccccgaaaca gagctgcctg ctgtgttaca gcctgtcgcc
     5221 gaagctatgg atgctattgc cgccgccgac ctcagtcaaa ccagcggctt cggcccattc
     5281 gggccccaag gcatcggcca gtacacaacc tggcgggatt tcatttgcgc cattgctgat
     5341 ccccatgtct accactggca gaccgtgatg gacgacaccg tgtccgccag cgtagctcaa
     5401 gccctggacg aactgatgct gtgggccgaa gactgtcccg aggtgcgcca cctcgtccat
     5461 gccgacttcg gcagcaacaa cgtcctgacc gacaacggcc gcatcaccgc cgtaatcgac
     5521 tggtccgaag ctatgttcgg ggacagtcag tacgaggtgg ccaacatctt cttctggcgg
     5581 ccctggctgg cttgcatgga gcagcagact cgctacttcg agcgccggca tcccgagctg
     5641 gccggcagcc ctcgtctgcg agcctacatg ctgcgcatcg gcctggatca gctctaccag
     5701 agcctcgtgg acggcaactt cgacgatgct gcctgggctc aaggccgctg cgatgccatc
     5761 gtccgcagcg gggccggcac cgtcggtcgc acacaaatcg ctcgccggag cgcagccgta
     5821 tggaccgacg gctgcgtcga ggtgctggcc gacagcggca accgccggcc cagtacacga
     5881 ccgcgcgcta aggaggtagg tcgagtttag actctagcca tcacgatggc cgcaataaaa
     5941 tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gcgataagga
     6001 tcctctttgc gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg
     6061 gtcagcaccg tttctgcgga ctggctttct acgtaatggt ttcttagacg tcaggtggca
     6121 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     6181 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     6241 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     6301 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     6361 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6421 ccgaagaacg ttttccaatg atgagcactt tcaaagttct gctatgtggc gcggtattat
     6481 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     6541 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     6601 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacta
     6661 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     6721 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     6781 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     6841 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     6901 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     6961 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     7021 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     7081 cctcactgat taagcattgg taattcgaaa tgaccgacca agcgacgccc aaccggtatc
     7141 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     7201 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     7261 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     7321 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     7381 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     7441 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     7501 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     7561 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     7621 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     7681 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     7741 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     7801 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt
     7861 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7921 catgagatta tcaaaaagga tcttcaccta gatcctttta tagtccggaa atacaggaac
     7981 gcacgctgga tggcccttcg ctgggatggt gaaaccatga aaaatggcag cttcagtgga
     8041 ttaagtgggg gtaatgtggc ctgtaccctc tggttgcata ggtattcata cggttaaaat
     8101 ttatcaggcg cgattgcggc agtttttcgg gtggtttgtt gccattttta cctgtctgct
     8161 gccgtgatcg cgctgaacgc gttttagcgg tgcgtacaat taagggatta tggtaaatcc
     8221 acttactgtc tgccctcgta gccatcgaga taaaccgcag tactccggcc acgatgcgtc
     8281 cggcgtagag gatcgagatc t
//
