LOCUS       sequence-411262-        7196 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect expression of human WDR12 fragment T90 to G422. N-terminal 
            biotin acceptor peptide tag and C-terminal 6X His tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7196)
  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 7196)
  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: pFBD-BirA_WDR12:90-422
FEATURES             Location/Qualifiers
     source          1..7196
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..457
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(89..108)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     299..320
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        484..588
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(589..657,658..1449)
                     /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(807..826)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1620..2208
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2109..2128
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2362..2379
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2511..2735
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2802..3335)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3524..3529,3530..3546,3547..3552))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    4061..4109
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4134..4153
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4180..5145)
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             complement(4183..5145)
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     promoter        complement(5182..5291)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        5315..5406
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     5331..5348
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             5457..5501
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             6534..6551
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    6686..6820
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6723..6742)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6777..6796
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  6849..7014
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
       61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
      121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
      181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
      241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
      301 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
      361 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
      421 aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt tcggggaaat
      481 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
      541 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
      601 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
      661 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
      721 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
      781 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
      841 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
      901 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
      961 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     1021 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     1081 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     1141 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     1201 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     1261 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     1321 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     1381 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     1441 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     1501 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     1561 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1621 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1681 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     1741 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     1801 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     1861 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     1921 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     1981 tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg
     2041 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     2101 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     2161 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     2221 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2281 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2341 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     2401 cggtattttc tccttacgca tctgtgcggt atttcacacc gcagaccagc cgcgtaacct
     2461 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga
     2521 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag
     2581 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt
     2641 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga
     2701 ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac
     2761 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     2821 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     2881 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     2941 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     3001 gcgagatcat agatatagat ctcactacgc ggctgctcaa acctgggcag aacgtaagcc
     3061 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     3121 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
     3181 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     3241 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     3301 ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata acatcaaaca
     3361 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacaaaaaa
     3421 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa
     3481 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca
     3541 ggcttatgtc aactgggttc gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac
     3601 cttgggcagc agcgaagtcg aggcatttct gtcctggctg gcgaacgagc gcaaggtttc
     3661 ggtctccacg catcgtcagg cattggcggc cttgctgttc ttctacggca aggtgctgtg
     3721 cacggatctg ccctggcttc aggagatcgg tagacctcgg ccgtcgcggc gcttgccggt
     3781 ggtgctgacc ccggatgaag tggttcgcat cctcggtttt ctggaaggcg agcatcgttt
     3841 gttcgcccag gactctagct atagttctag tggttggcct acgtacccgt agtggctatg
     3901 gcagggcttg ccgccccgac gttggctgcg agccctgggc cttcacccga acttgggggt
     3961 tggggtgggg aaaaggaaga aacgcgggcg tattggtccc aatggggtct cggtggggta
     4021 tcgacagagt gccagccctg ggaccgaacc ccgcgtttat gaacaaacga cccaacaccc
     4081 gtgcgtttta ttctgtcttt ttattgccgt catagcgcgg gttccttccg gtattgtctc
     4141 cttccgtgtt tcagttagcc tcccccatct cccggtactt catttttctg cactacgcag
     4201 ggatatttca ccgcccatcc agggttttat tattccatcc tgctcaagta ataaagcccc
     4261 ctgtttgtct attccgcgtg aaatgccaaa tatttcttta tcaccaatga taagtttcac
     4321 tgggcgatta ataaaattat ccagcttttc ccagcgcgac agataaggtg ccaatccttc
     4381 ttgttcgaag agttccaacg cagcacgtaa ttcacgtatt agcatggccg ccaacgtatt
     4441 acgatcgaga ttgatccccg cttcctgcag cgtgatccac ccctgattaa cgacactctc
     4501 ttcaacacgg cgcattgcca tgttgatccc ggctccaatg actatttgcg ccgcatcgcc
     4561 agttttgcca gtcagctcca ccagaatgcc tgccagcttg cgatcctgca gatagaggtc
     4621 attaggccat ttaacacgaa ctttatctgc acccagcttg cgtaatactt ccgccatcac
     4681 gataccgata accagactta aaccaatcgc cgccgccggg ccttgttcca gacgccagaa
     4741 catcgacaaa tataagtttg cgccaaaagg cgaaaaccat ttccgacccc ggcgaccacg
     4801 gccagcctgc tggtattctg caatgcaagc atcgcccgat ttaagctctc cgatacgatc
     4861 aagaaggtac tgattcgtgg agtcaatcac tggcagcacg gctacactac cgccatccag
     4921 ctgacccaat atctgtttag cattaagtaa ctggataggc tcaggcaggc tgtatccttt
     4981 acccggaacg gtaaagacat caacgcccca gtcacgcagt gtctgaatgt gtttattaat
     5041 agccgcccgg ctcattccca gcgtttcacc caactgctcg ccagagtgaa attcaccgtt
     5101 cgctaacagg gcaatcaatt tcagtggcac ggtgttatcc ttcatggctc gagatcccgg
     5161 gtgatcaagt cttcgtcgag tgattgtaaa taaaatgtaa tttacagtat agtattttaa
     5221 ttaatataca aatgatttga taataattct tatttaacta taatatattg tgttgggttg
     5281 aattaaaggt ccgtatactc cggaatatta atagatcatg gagataatta aaatgataac
     5341 catctcgcaa ataaataagt attttactgt tttcgtaaca gttttgtaat aaaaaaacct
     5401 ataaatattc cggattattc ataccgtccc accatcgggc gcggatcccg atgagcggcc
     5461 tgaacgatat ttttgaagcg cagaaaattg aatggcatga aggcagcgct ggaggttcag
     5521 gtactgcacc ccagccagag caatgcatgt tccatgatga ctggatcagt tcaattaaag
     5581 gggcagagga atggatcttg actggttctt atgataagac ttctcggatc tggtccttgg
     5641 aaggaaagtc aataatgaca attgtgggac atacggatgt tgtaaaagat gtggcctggg
     5701 tgaaaaaaga tagtttgtcc tgcttattat tgagtgcttc tatggatcag actattctct
     5761 tatgggagtg gaatgtagag agaaacaaag tgaaagccct acactgctgt agaggtcatg
     5821 ctggaagtgt agattctata gctgttgatg gctcaggaac taaattttgc agtggctcct
     5881 gggataagat gctaaagatc tggtctacag tccctacaga tgaagaagat gaaatggagg
     5941 agtccacaaa tcgaccaaga aagaaacaga agacagaaca gttgggacta acaaggactc
     6001 ccatagtgac cctctctggc cacatggagg cagtttcctc agttctgtgg tcagatgctg
     6061 aagaaatctg cagtgcatct tgggaccata caattagagt gtgggatgtt gagtctggca
     6121 gtcttaagtc aactttgaca ggaaataaag tgtttaattg tatttcctat tctccacttt
     6181 gtaaacgttt agcatctgga agcacagata ggcatatcag actgtgggat ccccgaacta
     6241 aagatggttc tttggtgtcg ctgtccctaa cgtcacatac tggttgggtg acatcagtaa
     6301 aatggtctcc tacccatgaa cagcagctga tttcaggatc tttagataac attgttaagc
     6361 tgtgggatac aagaagttgt aaggctcctc tctatgatct ggctgctcat gaagacaaag
     6421 ttctgagtgt agactggaca gacacagggc tacttctgag tggaggagca gacaataaat
     6481 tgtattccta cagatattca cctaccactt cccatgttgg gggcggaagt ggacatcacc
     6541 accatcatca ctgatgaagc ttgtcgagaa gtactagagg atcataatca gccataccac
     6601 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     6661 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     6721 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     6781 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc tgatcactgc ttgagcctag
     6841 gagatccgaa ccagataagt gaaatctagt tccaaactat tttgtcattt ttaattttcg
     6901 tattagctta cgacgctaca cccagttccc atctattttg tcactcttcc ctaaataatc
     6961 cttaaaaact ccatttccac ccctcccagt tcccaactat tttgtccgcc cacagcgggg
     7021 catttttctt cctgttatgt ttttaatcaa acatcctgcc aactccatgt gacaaaccgt
     7081 catcttcggc tactttttct ctgtcacaga atgaaaattt ttctgtcatc tcttcgttat
     7141 taatgtttgt aattgactga atatcaacgc ttatttgcag cctgaatggc gaatgg
//
