LOCUS       sequence-346871-        5693 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Chlamydomonas reinhardtii RSP9 recombinant protein in 
            insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5693)
  AUTHORS   Grossman-Haham I, Coudray N, Yu Z, Wang F, Zhang N, Bhabha G, Vale 
            RD
  TITLE     Structure of the radial spoke head and insights into its role in 
            mechanoregulation of ciliary beating.
  JOURNAL   Nat Struct Mol Biol. 2021 Jan;28(1):20-28. doi: 
            10.1038/s41594-020-00519-9. Epub 2020 Dec 14.
  PUBMED    33318704
REFERENCE   2  (bases 1 to 5693)
  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: 
            "10.1038/s41594-020-00519-9"; journalName: "Nat Struct Mol Biol"; 
            date: "2021-01"; volume: "28"; issue: "1"; pages: "20-28"
COMMENT     Sequence Label: pLIB-RSP9
FEATURES             Location/Qualifiers
     source          1..5693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(512..517,518..534,535..540))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(936..969)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1017..1108
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1033..1050
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    2090..2224
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2127..2146)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2181..2200
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2334..2499
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2683..3137
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2770..2789)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2979..3000
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3163..3267
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3268..3336,3337..4128)
                     /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(3486..3505)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4299..4887
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4788..4807
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5041..5058
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  5192..5416
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(join(5483..5693,1..323))
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 atatagatct cactacgcgg ctgctcaaac ttgggcagaa cgtaagccgc gagagcgcca
       61 acaaccgctt cttggtcgaa ggcagcaagc gcgatgaatg tcttactacg gagcaagttc
      121 ccgaggtaat cggagtccgg ctgatgttgg gagtaggtgg ctacgtctcc gaactcacga
      181 ccgaaaagat caagagcagc ccgcatggat ttgacttggt cagggccgag cctacatgtg
      241 cgaatgatgc ccatacttga gccacctaac tttgttttag ggcgactgcc ctgctgcgta
      301 acatcgttgc tgctgcgtaa catcgttgct gctccataac atcaaacatc gacccacggc
      361 gtaacgcgct tgctgcttgg atgcccgagg catagactgt acaaaaaaac agtcataaca
      421 agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca
      481 gttgcgtgag cgcatacgct acttgcatta cagtttacga accgaacagg cttatgtcaa
      541 ctgggtttgt gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct tgggcagcag
      601 cgaagtcgag gcatttctgt cctggctggc gaacgagcgc aaggtttcgg tctccacgca
      661 tcgtcaggca ttggcggcct tgctgttctt ctacggcaag gtgctgtgca cggatctgcc
      721 cttgcttcag gagatcggta gacctcggcc gtcgcggcgc ttgccggtgg tgctgacccc
      781 ggatgaagtg gttcgcatcc tcggttttct ggaaggcgag catcgtttgt tcgcccagga
      841 ctctagctat agttctagtg gttggctaca gctttgtttg tactatcaac aggttgaact
      901 gctgatcaac agatcctcta cgcggccgcg gtaccataac ttcgtatagc atacattata
      961 cgaagttatc tggttttact agtatcgatt cgcgacctac tccggaatat taatagatca
     1021 tggagataat taaaatgata accatctcgc aaataaataa gtattttact gttttcgtaa
     1081 cagttttgta ataaaaaaac ctataaatat tccggattat tcataccgtc ccaccatcgg
     1141 gcgcggatcc atggttcagt tggagcctaa tatcaccctt gtccttaagc atctggcatc
     1201 ttgcggtgca gtcgttagcg cggagcagca ggccgcattg gatcattcta tcccaattaa
     1261 gagaatcgag gcgggtctgc gtagccttac gctttgggga agattgacga cccttaatgg
     1321 caaggattat cttgtcgccg agggttacaa cgtcgcctca agtaaagaag gagcagctgt
     1381 ttacgaaact aaatacttct attcccaaga tggcgctcgt tggagtgact tgcagccagt
     1441 agattccgaa actgccacac gctgcgcgag gatcaaggga atgctcagtg gtgatcccgc
     1501 taaaaactac gagctggagg agaaggatcc aaacgcccct gagccgtccc cggaagccga
     1561 agaagaagtg aagccgcttg tcttccaaat accggaactg gccgtgctca ggtgccgcgt
     1621 ggacgctatc gccacagcaa cttctgtaat cccgactgat tctactatcc tgaacgcagc
     1681 tagtcaggtt gtaccgaacc gcttgttcgc aggtgcagca tacccggaaa agcttgagtc
     1741 ctatcagcac agattctctc tgccaggttc cggtgttaca ctcagccaag acctgcgcgg
     1801 tacttgggct gtccaatacg atgcctttaa gggcgtcgcg caggtcagat ccctgctttt
     1861 ccctggatat tttttttatt atgccgcgaa tgagctgact tggggatcgc tttatgttgg
     1921 agatggactt agaaataatg atctgatatt tatgctctaa aagcttgtcg agaagtacta
     1981 gaggatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     2041 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     2101 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     2161 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     2221 gatctgatca ctgcttgagc ctagaagatc cggctgctaa caaagcccga aaggaagctg
     2281 agttggctgc tgccaccgct gagcaataac tatcataacc cctaggagat ccgaaccaga
     2341 taagtgaaat ctagttccaa actattttgt catttttaat tttcgtatta gcttacgacg
     2401 ctacacccag ttcccatcta ttttgtcact cttccctaaa taatccttaa aaactccatt
     2461 tccacccctc ccagttccca actattttgt ccgcccacag cggggcattt ttcttcctgt
     2521 tatgttttta atcaaacatc ctgccaactc catgtgacaa accgtcatct tcggctactt
     2581 tttctctgtc acagaatgaa aatttttctg tcatctcttc gttattaatg tttgtaattg
     2641 actgaatatc aacgcttatt tgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     2701 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     2761 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     2821 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     2881 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     2941 tttttcgccc tttgacgttg gagtccacgt tcttaatagt ggactcttgt tccaaactgg
     3001 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3061 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3121 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3181 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3241 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3301 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3361 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3421 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     3481 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     3541 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     3601 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     3661 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     3721 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     3781 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     3841 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     3901 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     3961 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4021 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4081 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4141 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4201 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4261 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4321 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4381 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4441 actattcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4501 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4561 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4621 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4681 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4741 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4801 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4861 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4921 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     4981 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     5041 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     5101 ctgtgcggta tttcacaccg catagaccag ccgcgtaacc tggcaaaatc ggttacggtt
     5161 gagtaataaa tggatgccct gcgtaagcgg gtgtgggcgg acaataaagt cttaaactga
     5221 acaaaataga tctaaactat gacaataaag tcttaaacta gacagaatag ttgtaaactg
     5281 aaatcagtcc agttatgctg tgaaaaagca tactggactt ttgttatggc taaagcaaac
     5341 tcttcatttt ctgaagtgca aattgcccgt cgtattaaag aggggcgtgg ccaagggcat
     5401 ggtaaagact atattcgcgg cgttgtgaca atttaccgaa caactccgcg gccgggaagc
     5461 cgatctcggc ttgaacgaat tgttaggtgg cggtacttgg gtcgatatca aagtgcatca
     5521 cttcttcccg tatgcccaac tttgtataga gagccactgc gggatcgtca ccgtaatctg
     5581 cttgcacgta gatcacataa gcaccaagcg cgttggcctc atgcttgagg agattgatga
     5641 gcgcggtggc aatgccctgc ctccggtgct cgccggagac tgcgagatca tag
//
