LOCUS       sequence-346868-        5498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Chlamydomonas reinhardtii RSP11 recombinant protein in
            insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5498)
  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 5498)
  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-RSP11
FEATURES             Location/Qualifiers
     source          1..5498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(102..107,108..124,125..130))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(526..559)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        607..698
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     623..640
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    1485..1619
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1522..1541)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1576..1595
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1729..1894
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2078..2532
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2165..2184)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2374..2395
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2558..2662
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2663..2731,2732..3523)
                     /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(2881..2900)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3694..4282
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4183..4202
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4436..4453
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  4587..4811
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(4878..5411)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 agtcataaca agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg
       61 ttctggacca gttgcgtgag cgcatacgct acttgcatta cagtttacga accgaacagg
      121 cttatgtcaa ctgggtttgt gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct
      181 tgggcagcag cgaagtcgag gcatttctgt cctggctggc gaacgagcgc aaggtttcgg
      241 tctccacgca tcgtcaggca ttggcggcct tgctgttctt ctacggcaag gtgctgtgca
      301 cggatctgcc cttgcttcag gagatcggta gacctcggcc gtcgcggcgc ttgccggtgg
      361 tgctgacccc ggatgaagtg gttcgcatcc tcggttttct ggaaggcgag catcgtttgt
      421 tcgcccagga ctctagctat agttctagtg gttggctaca gctttgtttg tactatcaac
      481 aggttgaact gctgatcaac agatcctcta cgcggccgcg gtaccataac ttcgtatagc
      541 atacattata cgaagttatc tggttttact agtatcgatt cgcgacctac tccggaatat
      601 taatagatca tggagataat taaaatgata accatctcgc aaataaataa gtattttact
      661 gttttcgtaa cagttttgta ataaaaaaac ctataaatat tccggattat tcataccgtc
      721 ccaccatcgg gcgcggatcc atggatgtgg aacccatatt ttgcgctgag caaatagtca
      781 tcccacacaa tttggccgat atactgaaag catacacaaa agaagtcatc cgtcgccaac
      841 ctaccgacct catagccttc tcagcgaaat acttcaccaa cctggccaac gtggcgtctg
      901 gtgtttctaa ttcttcagcg cctgctaaag aacagttgcg ccaggtgtat acgcgtggtg
      961 gttccggtgg cgcaacactt actgagagcc aggtgaccgg attgtgtcaa caggcgggta
     1021 ttgccgatgc ggttgtagca aaagtgatgg aagttggagc attcacgcca gcggctgtag
     1081 atctcagtaa atttgttttc ttgtgtttgg ccatgagttg cgaggatttc aacagggttt
     1141 gcatgggtgt gttcgatgtg ttttcagata atggaagcct gcccgctcaa gatcttctta
     1201 cgctgattgc ccatcttgga ccggacatgg atcccgaagt tacgccggct tttctcgacg
     1261 ccgttgcggc ggaactcccg gcaggcggtg gtgcagtgac gtatatggaa ctttgcgaag
     1321 caccctctct gaagcccaag ttgggtttgt cctaaaagct tgtcgagaag tactagagga
     1381 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     1441 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     1501 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     1561 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct
     1621 gatcactgct tgagcctaga agatccggct gctaacaaag cccgaaagga agctgagttg
     1681 gctgctgcca ccgctgagca ataactatca taacccctag gagatccgaa ccagataagt
     1741 gaaatctagt tccaaactat tttgtcattt ttaattttcg tattagctta cgacgctaca
     1801 cccagttccc atctattttg tcactcttcc ctaaataatc cttaaaaact ccatttccac
     1861 ccctcccagt tcccaactat tttgtccgcc cacagcgggg catttttctt cctgttatgt
     1921 ttttaatcaa acatcctgcc aactccatgt gacaaaccgt catcttcggc tactttttct
     1981 ctgtcacaga atgaaaattt ttctgtcatc tcttcgttat taatgtttgt aattgactga
     2041 atatcaacgc ttatttgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     2101 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2161 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2221 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2281 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2341 cgccctttga cgttggagtc cacgttctta atagtggact cttgttccaa actggaacaa
     2401 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     2461 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     2521 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2581 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2641 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2701 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2761 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2821 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     2881 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     2941 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3001 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3061 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3121 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3181 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3241 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3301 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3361 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3421 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3481 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3541 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3601 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3661 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3721 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3781 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactat
     3841 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3901 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3961 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4021 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4081 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4141 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4201 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4261 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4321 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     4381 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     4441 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     4501 cggtatttca caccgcatag accagccgcg taacctggca aaatcggtta cggttgagta
     4561 ataaatggat gccctgcgta agcgggtgtg ggcggacaat aaagtcttaa actgaacaaa
     4621 atagatctaa actatgacaa taaagtctta aactagacag aatagttgta aactgaaatc
     4681 agtccagtta tgctgtgaaa aagcatactg gacttttgtt atggctaaag caaactcttc
     4741 attttctgaa gtgcaaattg cccgtcgtat taaagagggg cgtggccaag ggcatggtaa
     4801 agactatatt cgcggcgttg tgacaattta ccgaacaact ccgcggccgg gaagccgatc
     4861 tcggcttgaa cgaattgtta ggtggcggta cttgggtcga tatcaaagtg catcacttct
     4921 tcccgtatgc ccaactttgt atagagagcc actgcgggat cgtcaccgta atctgcttgc
     4981 acgtagatca cataagcacc aagcgcgttg gcctcatgct tgaggagatt gatgagcgcg
     5041 gtggcaatgc cctgcctccg gtgctcgccg gagactgcga gatcatagat atagatctca
     5101 ctacgcggct gctcaaactt gggcagaacg taagccgcga gagcgccaac aaccgcttct
     5161 tggtcgaagg cagcaagcgc gatgaatgtc ttactacgga gcaagttccc gaggtaatcg
     5221 gagtccggct gatgttggga gtaggtggct acgtctccga actcacgacc gaaaagatca
     5281 agagcagccc gcatggattt gacttggtca gggccgagcc tacatgtgcg aatgatgccc
     5341 atacttgagc cacctaactt tgttttaggg cgactgccct gctgcgtaac atcgttgctg
     5401 ctgcgtaaca tcgttgctgc tccataacat caaacatcga cccacggcgt aacgcgcttg
     5461 ctgcttggat gcccgaggca tagactgtac aaaaaaac
//
