LOCUS       sequence-426071-        5684 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect expression of human CDC20 fragment S161 to P477. N-terminal 
            6X His tag with TEV cleavage..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5684)
  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 5684)
  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: pFBOH-MHL_CDC20:161-477
FEATURES             Location/Qualifiers
     source          1..5684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      55..509
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(142..161)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     351..372
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        535..639
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(640..708,709..1500)
                     /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(858..877)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1671..2259
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2160..2179
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2413..2430
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2564..2788
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2855..3388)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3558..3563,3564..3580,3581..3586))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3938..4029
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3954..3971
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4088..4105
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4118..4138
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    5227..5361
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5264..5283)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5318..5337
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  5390..5555
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 tgtttgtaat tgactgaata tcaacgctta tttgcagcct gaatggcgaa tgggacgcgc
       61 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
      121 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
      181 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
      241 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
      301 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttcttaata gtggactctt
      361 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
      421 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
      481 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
      541 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
      601 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
      661 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
      721 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
      781 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
      841 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
      901 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
      961 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     1021 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     1081 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     1141 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     1201 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     1261 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     1321 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     1381 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     1441 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     1501 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     1561 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     1621 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1681 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1741 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     1801 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     1861 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     1921 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     1981 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2041 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2101 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2161 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2221 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     2281 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     2341 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     2401 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     2461 ctccttacgc atctgtgcgg tatttcacac cgcatagacc agccgcgtaa cctggcaaaa
     2521 tcggttacgg ttgagtaata aatggatgcc ctgcgtaagc gggtgtgggc ggacaataaa
     2581 gtcttaaact gaacaaaata gatctaaact atgacaataa agtcttaaac tagacagaat
     2641 agttgtaaac tgaaatcagt ccagttatgc tgtgaaaaag catactggac ttttgttatg
     2701 gctaaagcaa actcttcatt ttctgaagtg caaattgccc gtcgtattaa agaggggcgt
     2761 ggccaagggc atggtaaaga ctatattcgc ggcgttgtga caatttaccg aacaactccg
     2821 cggccgggaa gccgatctcg gcttgaacga attgttaggt ggcggtactt gggtcgatat
     2881 caaagtgcat cacttcttcc cgtatgccca actttgtata gagagccact gcgggatcgt
     2941 caccgtaatc tgcttgcacg tagatcacat aagcaccaag cgcgttggcc tcatgcttga
     3001 gcagattgat gagcgcggtg gcaatgccct gcctccggtg ctcgccggag actgcgagat
     3061 catagatata gatctcacta cgcggctgct caaacttggg cagaacgtaa gccgcgagag
     3121 cgccaacaac cgcttcttgg tcgaaggcag caagcgcgat gaatgtctta ctacggagca
     3181 agttcccgag gtaatcggag tccggctgat gttgggagta ggtggctacg tctccgaact
     3241 cacgaccgaa aagatcaaga gcagcccgca tggatttgac ttggtcaggg ccgagcctac
     3301 atgtgcgaat gatgcccata cttgagccac ctaactttgt tttagggcga ctgccctgct
     3361 gcgtaacatc gttgctgctc cataacatca aacatcgacc cacggcgtaa cgcgcttgct
     3421 gcttggatgc ccgaggcata gactgtacaa aaaaacagtc ataacaagcc atgaaaaccg
     3481 ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg cgtgagcgca
     3541 tacgctactt gcattacagt ttacgaaccg aacaggctta tgtcaactgg gttcgtgcct
     3601 tcatccgttt ccacggtgtg cgtcacccgg caaccttggg cagcagcgaa gtcgaggcat
     3661 ttctgtcctg gctggcgaac gagcgcaagg tttcggtctc cacgcatcgt caggcattgg
     3721 cggccttgct gttcttctac ggcaaggtgc tgtgcacgga tctgccctgg cttcaggaga
     3781 tcggaagacc tcggccgtcg cggcgcttgc cggtggtgct gaccccggat gaagtggttc
     3841 gcatcctcgg ttttctggaa ggcgagcatc gtttgttcgc ccaggactct agctatagtt
     3901 ctagtggttg gctacgtata ctccggaata ttaatagatc atggagataa ttaaaatgat
     3961 aaccatctcg caaataaata agtattttac tgttttcgta acagttttgt aataaaaaaa
     4021 cctataaata ttccggatta ttcataccgt cccaccatcg ggcgcggatc tcggtccgaa
     4081 aaccatgcat catcatcatc atcacagcag cggcagagaa aacttgtatt tccagggcag
     4141 ccggaagacc tgccgttaca ttccttccct gccagaccgt atcctggatg cgcctgaaat
     4201 ccgaaatgac tattacctga accttgtgga ttggagttct gggaatgtac tggccgtggc
     4261 actggacaac agtgtgtacc tgtggagtgc aagctctggt gacatcctgc agcttttgca
     4321 aatggagcag cctggggaat atatatcctc tgtggcctgg atcaaagagg gcaactactt
     4381 ggctgtgggc accagcagtg ctgaggtgca gctatgggat gtgcagcagc agaaacggct
     4441 tcgaaatatg accagtcact ctgcccgagt gggctcccta agctggaaca gctatatcct
     4501 gtccagtggt tcacgttctg gccacatcca ccaccatgat gttcgggtag cagaacacca
     4561 tgtggccaca ctgagtggcc acagccagga agtgtgtggg ctgcgctggg ccccagatgg
     4621 acgacatttg gccagtggtg gtaatgataa cttggtcaat gtgtggccta gtgctcctgg
     4681 agagggtggc tgggttcctc tgcagacatt cacccagcat caaggggctg tcaaggccgt
     4741 agcatggtgt ccctggcagt ccaatgtcct ggcaacagga gggggcacca gtgatcgaca
     4801 cattcgcatc tggaatgtgt gctctggggc ctgtctgagt gccgtggatg cccattccca
     4861 ggtgtgctcc atcctctggt ctccccatta caaggagctc atctcaggcc atggctttgc
     4921 acagaaccag ctagttattt ggaagtaccc aaccatggcc aaggtggctg aactcaaagg
     4981 tcacacatcc cgggtcctga gtctgaccat gagcccagat ggggccacag tggcatccgc
     5041 agcagcagat gagaccctga ggctatggcg ctgttttgag ttggaccctt gatgacgaag
     5101 cttgtcgaga agtactagag gatcataatc agccatacca catttgtaga ggttttactt
     5161 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     5221 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5281 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5341 gtatcttatc atgtctggat ctgatcactg atatcgccta ggagatccga accagataag
     5401 tgaaatctag ttccaaacta ttttgtcatt tttaattttc gtattagctt acgacgctac
     5461 acccagttcc catctatttt gtcactcttc cctaaataat ccttaaaaac tccatttcca
     5521 cccctcccag ttcccaacta ttttgtccgc ccacagcggg gcatttttct tcctgttatg
     5581 tttttaatca aacatcctgc caactccatg tgacaaaccg tcatcttcgg ctactttttc
     5641 tctgtcacag aatgaaaatt tttctgtcat ctcttcgtta ttaa
//
