LOCUS       sequence-426046-        6074 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect expression of full-length human GRWD1. N-terminal 6X His tag 
            with thrombin protease cleavage site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6074)
  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 6074)
  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-LIC_GRWD1:1-446
FEATURES             Location/Qualifiers
     source          1..6074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      342..930
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     831..850
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1084..1101
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  1235..1459
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1526..2059)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2229..2234,2235..2251,2252..2257))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        2609..2700
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     2625..2642
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             2768..2785
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2795..2812
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     polyA_signal    4288..4422
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4325..4344)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4379..4398
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4451..4616
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4800..5254
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4887..4906)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5096..5117
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5280..5384
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5385..5453,5454..6074,1..171)
                     /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(5603..5622)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
       61 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
      121 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
      181 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
      241 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
      301 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
      361 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
      421 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
      481 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
      541 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
      601 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
      661 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
      721 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
      781 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
      841 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
      901 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
      961 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     1021 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     1081 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     1141 catctgtgcg gtatttcaca ccgcatagac cagccgcgta acctggcaaa atcggttacg
     1201 gttgagtaat aaatggatgc cctgcgtaag cgggtgtggg cggacaataa agtcttaaac
     1261 tgaacaaaat agatctaaac tatgacaata aagtcttaaa ctagacagaa tagttgtaaa
     1321 ctgaaatcag tccagttatg ctgtgaaaaa gcatactgga cttttgttat ggctaaagca
     1381 aactcttcat tttctgaagt gcaaattgcc cgtcgtatta aagaggggcg tggccaaggg
     1441 catggtaaag actatattcg cggcgttgtg acaatttacc gaacaactcc gcggccggga
     1501 agccgatctc ggcttgaacg aattgttagg tggcggtact tgggtcgata tcaaagtgca
     1561 tcacttcttc ccgtatgccc aactttgtat agagagccac tgcgggatcg tcaccgtaat
     1621 ctgcttgcac gtagatcaca taagcaccaa gcgcgttggc ctcatgcttg agcagattga
     1681 tgagcgcggt ggcaatgccc tgcctccggt gctcgccgga gactgcgaga tcatagatat
     1741 agatctcact acgcggctgc tcaaacttgg gcagaacgta agccgcgaga gcgccaacaa
     1801 ccgcttcttg gtcgaaggca gcaagcgcga tgaatgtctt actacggagc aagttcccga
     1861 ggtaatcgga gtccggctga tgttgggagt aggtggctac gtctccgaac tcacgaccga
     1921 aaagatcaag agcagcccgc atggatttga cttggtcagg gccgagccta catgtgcgaa
     1981 tgatgcccat acttgagcca cctaactttg ttttagggcg actgccctgc tgcgtaacat
     2041 cgttgctgct ccataacatc aaacatcgac ccacggcgta acgcgcttgc tgcttggatg
     2101 cccgaggcat agactgtaca aaaaaacagt cataacaagc catgaaaacc gccactgcgc
     2161 cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc atacgctact
     2221 tgcattacag tttacgaacc gaacaggctt atgtcaactg ggttcgtgcc ttcatccgtt
     2281 tccacggtgt gcgtcacccg gcaaccttgg gcagcagcga agtcgaggca tttctgtcct
     2341 ggctggcgaa cgagcgcaag gtttcggtct ccacgcatcg tcaggcattg gcggccttgc
     2401 tgttcttcta cggcaaggtg ctgtgcacgg atctgccctg gcttcaggag atcggaagac
     2461 ctcggccgtc gcggcgcttg ccggtggtgc tgaccccgga tgaagtggtt cgcatcctcg
     2521 gttttctgga aggcgagcat cgtttgttcg cccaggactc tagctatagt tctagtggtt
     2581 ggctacgtat actccggaat attaatagat catggagata attaaaatga taaccatctc
     2641 gcaaataaat aagtatttta ctgttttcgt aacagttttg taataaaaaa acctataaat
     2701 attccggatt attcataccg tcccaccatc gggcgcggat ctcggtccga aaaccatggg
     2761 cagcagccat catcatcatc atcacagcag cggcctggtt ccgcgtggta gtatggcggc
     2821 gcgcaagggt cggcgtcgca cgtgtgaaac cggggaaccc atggaagccg agtccggcga
     2881 cacaagttcc gagggcccgg cccaggtcta cctgcccggc cgggggccgc cgctacgcga
     2941 aggggaggag ctggtcatgg acgaggaggc ctatgtgctc taccaccgag cgcagactgg
     3001 cgccccctgt ctcagctttg acatagtccg ggatcacctg ggagacaacc ggacagagct
     3061 tcctcttaca ctttacttgt gtgctgggac ccaggctgag agcgcccaga gcaacagact
     3121 gatgatgctt cggatgcaca atctgcatgg gacaaagccc ccaccctcag agggcagtga
     3181 tgaagaagaa gaggaggaag atgaagagga tgaagaagag cggaaacctc agctggagct
     3241 ggccatggtg ccccactatg gtggcatcaa ccgagttcgg gtgtcatggc tgggtgaaga
     3301 gcctgtggct ggggtgtggt cagagaaggg ccaggtggag gtgtttgcgc tgcggcggct
     3361 tctgcaggtg gtggaggagc cccaggccct ggcagccttc ctccgggatg agcaggccca
     3421 aatgaagccc atcttctcct tcgctggaca catgggcgag ggctttgccc ttgactggtc
     3481 cccccgggtg accggtcgcc tgctgaccgg tgactgtcaa aagaacatcc acctctggac
     3541 acctacggac ggcggctcct ggcacgtgga ccagcggcca ttcgtgggcc acacacgctc
     3601 tgtggaggac ctgcagtggt caccgactga gaacacggtg tttgcctcct gctcagctga
     3661 cgcctccatc cgcatctggg acatccgggc agcccccagc aaggcctgca tgctcaccac
     3721 agccaccgcc catgatgggg acgtcaatgt catcagctgg agccgccggg agcccttcct
     3781 gctcagtggc ggggatgatg gggccctcaa gatctgggac cttcggcagt tcaagtctgg
     3841 ttccccagtg gccaccttca agcagcacgt ggcccccgtg acctccgtcg agtggcaccc
     3901 ccaggacagc ggggtctttg cagcctcggg tgcagaccac cagatcacac agtgggacct
     3961 ggcagtggag cgggaccctg aggcgggcga cgtggaggcc gaccccggac tggccgacct
     4021 cccgcagcag ctgctgttcg tgcaccaggg cgagaccgag ctgaaggagc tgcactggca
     4081 cccgcagtgc ccagggctcc tggtcagcac ggcgctgtca ggcttcacca tcttccgcac
     4141 catcagcgtc tgatgacgaa gcttgtcgag aagtactaga ggatcataat cagccatacc
     4201 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     4261 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     4321 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     4381 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tctgatcact gatatcgcct
     4441 aggagatccg aaccagataa gtgaaatcta gttccaaact attttgtcat ttttaatttt
     4501 cgtattagct tacgacgcta cacccagttc ccatctattt tgtcactctt ccctaaataa
     4561 tccttaaaaa ctccatttcc acccctccca gttcccaact attttgtccg cccacagcgg
     4621 ggcatttttc ttcctgttat gtttttaatc aaacatcctg ccaactccat gtgacaaacc
     4681 gtcatcttcg gctacttttt ctctgtcaca gaatgaaaat ttttctgtca tctcttcgtt
     4741 attaatgttt gtaattgact gaatatcaac gcttatttgc agcctgaatg gcgaatggga
     4801 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4861 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4921 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4981 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     5041 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct taatagtgga
     5101 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     5161 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     5221 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     5281 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     5341 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     5401 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5461 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     5521 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     5581 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5641 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5701 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5761 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5821 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5881 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5941 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     6001 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     6061 gctggtttat tgct
//
