LOCUS       sequence-426130-        5894 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Insect expression of human LRRK2 fragment T2141 to E2527. N-terminal
            6X His tag with TEV cleavage..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5894)
  AUTHORS   Ackloo S, Li F, Szewczyk M, Seitova A, Loppnau P, Zeng H, Xu J, 
            Ahmad S, Arnautova Y, Baghaie A, Beldar S, Bolotokova A, Centrella 
            P, Chau I, Clark M, Cuozzo J, Dehghani-Tafti S, Disch J, Dong A, 
            Dumas A, Feng J, Ghiabi P, Gibson E, Gilmer J, Goldman B, Green S, 
            Guie M, Guilinger J, Harms N, Herasymenko O, Houliston S, Hutchinson
            A, Kearnes S, Keefe A, Kimani S, Kramer T, Kutera M, Kwak H, Lento 
            C, Li Y, Liu J, Loup J, Machado R, Mulhern C, Perveen S, Righetto G,
            Riley P, Shrestha S, Sigel E, Silva M, Sintchak M, Slakman B, Taylor
            R, Thompson J, Torng W, Underkoffler C, Rechenberg M, Watson I, 
            Wilson D, Wolf E, Yadav M, Yazdi A, Zhang J, Zhang Y, Santhakumar V,
            Edwards A, Barsyte-Lovejoy D, Schapira M, Brown P, Halabelian L, 
            Arrowsmith C
  TITLE     A resource to enable chemical biology and drug discovery of WDR 
            Proteins
  JOURNAL   bioRxiv 2024.03.03.583197
REFERENCE   2  (bases 1 to 5894)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2024.03.03.583197"
COMMENT     Sequence Label: pFBOH-MHL_LRRK2:2141-2527
FEATURES             Location/Qualifiers
     source          1..5894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(205..224)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1018..1606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1507..1526
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1760..1777
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  1911..2135
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2202..2735)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2905..2910,2911..2927,2928..2933))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3285..3376
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3301..3318
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             3435..3452
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3465..3485
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    4784..4918
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4821..4840)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4875..4894
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4947..5112
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      5296..5750
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5383..5402)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5592..5613
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5776..5880
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5881..5894,1..55,56..847)
                     /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"
ORIGIN
        1 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
       61 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
      121 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
      181 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
      241 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
      301 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
      361 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
      421 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
      481 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
      541 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
      601 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
      661 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
      721 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
      781 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
      841 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
      901 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
      961 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     1021 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     1081 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     1141 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     1201 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     1261 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     1321 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     1381 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     1441 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     1501 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     1561 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     1621 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     1681 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     1741 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     1801 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct
     1861 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga
     1921 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag
     1981 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt
     2041 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga
     2101 ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac
     2161 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     2221 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     2281 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     2341 tgcttgagca gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     2401 gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc
     2461 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     2521 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
     2581 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     2641 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     2701 ccctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc
     2761 gcttgctgct tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccatg
     2821 aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt
     2881 gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt caactgggtt
     2941 cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc
     3001 gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag
     3061 gcattggcgg ccttgctgtt cttctacggc aaggtgctgt gcacggatct gccctggctt
     3121 caggagatcg gaagacctcg gccgtcgcgg cgcttgccgg tggtgctgac cccggatgaa
     3181 gtggttcgca tcctcggttt tctggaaggc gagcatcgtt tgttcgccca ggactctagc
     3241 tatagttcta gtggttggct acgtatactc cggaatatta atagatcatg gagataatta
     3301 aaatgataac catctcgcaa ataaataagt attttactgt tttcgtaaca gttttgtaat
     3361 aaaaaaacct ataaatattc cggattattc ataccgtccc accatcgggc gcggatctcg
     3421 gtccgaaaac catgcatcat catcatcatc acagcagcgg cagagaaaac ttgtatttcc
     3481 agggcacgag acgcatttta ttacctaaaa acgtaattgt tgaatgcatg gttgctacac
     3541 atcacaacag caggaatgca agcatttggc tgggctgtgg gcacaccgac agaggacagc
     3601 tctcatttct tgacttaaat actgaaggat acacttctga ggaagttgct gatagtagaa
     3661 tattgtgctt agccttggtg catcttcctg ttgaaaagga aagctggatt gtgtctggga
     3721 cacagtctgg tactctcctg gtcatcaata ccgaagatgg gaaaaagaga cataccctag
     3781 aaaagatgac tgattctgtc acttgtttgt attgcaattc cttttccaag caaagcaaac
     3841 aaaaaaattt tcttttggtt ggaaccgctg atggcaagtt agcaattttt gaagataaga
     3901 ctgttaagct taaaggagct gctcctttga agatactaaa tataggaaat gtcagtactc
     3961 cattgatgtg tttgagtgaa tccacaaatt caacggaaag aaatgtaatg tggggaggat
     4021 gtggcacaaa gattttctcc ttttctaatg atttcaccat tcagaaactc attgagacaa
     4081 gaacaagcca actgttttct tatgcagctt tcagtgattc caacatcata acagtggtgg
     4141 tagacactgc tctctatatt gctaagcaaa atagccctgt tgtggaagtg tgggataaga
     4201 aaactgaaaa actctgtgga ctaatagact gcgtgcactt tttaagggag gtaacggtaa
     4261 aagaaaacaa ggaatcaaaa cacaaaatgt cttattctgg gagagtgaaa accctctgcc
     4321 ttcagaagaa cactgctctt tggataggaa ctggaggagg ccatatttta ctcctggatc
     4381 tttcaactcg tcgacttata cgtgtaattt acaacttttg taattcggtc agagtcatga
     4441 tgacagcaca gctaggaagc cttaaaaatg tcatgctggt attgggctac aaccggaaaa
     4501 atactgaagg tacacaaaag cagaaagaga tacaatcttg cttgaccgtt tgggacatca
     4561 atcttccaca tgaagtgcaa aatttagaaa aacacattga agtgagaaaa gaattagctg
     4621 aaaaaatgag acgaacatct gttgagtgat gacgaagctt gtcgagaagt actagaggat
     4681 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     4741 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     4801 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4861 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatctg
     4921 atcactgata tcgcctagga gatccgaacc agataagtga aatctagttc caaactattt
     4981 tgtcattttt aattttcgta ttagcttacg acgctacacc cagttcccat ctattttgtc
     5041 actcttccct aaataatcct taaaaactcc atttccaccc ctcccagttc ccaactattt
     5101 tgtccgccca cagcggggca tttttcttcc tgttatgttt ttaatcaaac atcctgccaa
     5161 ctccatgtga caaaccgtca tcttcggcta ctttttctct gtcacagaat gaaaattttt
     5221 ctgtcatctc ttcgttatta atgtttgtaa ttgactgaat atcaacgctt atttgcagcc
     5281 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     5341 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     5401 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     5461 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     5521 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5581 cgttcttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     5641 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     5701 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     5761 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5821 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5881 atgagtattc aaca
//
