LOCUS       sequence-346869-        6467 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Chlamydomonas reinhardtii RSP5 recombinant protein in 
            insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6467)
  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 6467)
  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-RSP5
FEATURES             Location/Qualifiers
     source          1..6467
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(510..515,516..532,533..538))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(934..967)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1015..1106
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1031..1048
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    2862..2996
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2899..2918)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2953..2972
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3106..3271
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3455..3909
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3542..3561)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3751..3772
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3935..4039
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4040..4108,4109..4900)
                     /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(4258..4277)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5071..5659
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5560..5579
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5813..5830
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  5964..6188
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(join(6255..6467,1..321))
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 atagatctca ctacgcggct gctcaaactt gggcagaacg taagccgcga gagcgccaac
       61 aaccgcttct tggtcgaagg cagcaagcgc gatgaatgtc ttactacgga gcaagttccc
      121 gaggtaatcg gagtccggct gatgttggga gtaggtggct acgtctccga actcacgacc
      181 gaaaagatca agagcagccc gcatggattt gacttggtca gggccgagcc tacatgtgcg
      241 aatgatgccc atacttgagc cacctaactt tgttttaggg cgactgccct gctgcgtaac
      301 atcgttgctg ctgcgtaaca tcgttgctgc tccataacat caaacatcga cccacggcgt
      361 aacgcgcttg ctgcttggat gcccgaggca tagactgtac aaaaaaacag tcataacaag
      421 ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt
      481 tgcgtgagcg catacgctac ttgcattaca gtttacgaac cgaacaggct tatgtcaact
      541 gggtttgtgc cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg ggcagcagcg
      601 aagtcgaggc atttctgtcc tggctggcga acgagcgcaa ggtttcggtc tccacgcatc
      661 gtcaggcatt ggcggccttg ctgttcttct acggcaaggt gctgtgcacg gatctgccct
      721 tgcttcagga gatcggtaga cctcggccgt cgcggcgctt gccggtggtg ctgaccccgg
      781 atgaagtggt tcgcatcctc ggttttctgg aaggcgagca tcgtttgttc gcccaggact
      841 ctagctatag ttctagtggt tggctacagc tttgtttgta ctatcaacag gttgaactgc
      901 tgatcaacag atcctctacg cggccgcggt accataactt cgtatagcat acattatacg
      961 aagttatctg gttttactag tatcgattcg cgacctactc cggaatatta atagatcatg
     1021 gagataatta aaatgataac catctcgcaa ataaataagt attttactgt tttcgtaaca
     1081 gttttgtaat aaaaaaacct ataaatattc cggattattc ataccgtccc accatcgggc
     1141 gcggatccat gtccgagccc ggagaagagc ctgtagctgc gcccgctggc ccagcacccg
     1201 acccagtact gaatgaattg tatggatcgg agaggccagc ggtcgagctc cttccgggag
     1261 tgccactcag tcccatcgta aattcatgct ggcttcccgc ggatgcaaaa gcaatgctcg
     1321 cggaatcctg gatacccgtg cctccagagg atgcaggaga agaggcaggt cctccgcccc
     1381 cagcgtttga agcggccgcc ccggagtata atgaattggt ccgtcgcttg gcaaaaaccg
     1441 ctccattccg caaatggaac gaattgacta ttcaggcgaa acaacttgaa caggaagttg
     1501 cgggtctgaa aggaccggac gccgaagcga aacaagctga actcgaaaat gttaaagtcc
     1561 agatagctga tgctgaggcg gcggttgctg aggtgaaaca gagcttttct gacgatcctc
     1621 tctcacttac aggttggatg caggccctta ccgacttggc agacggaggt atgacgacat
     1681 ttgaggtatc tggccaagga tggccctact gctctctgag acagttgttt ggcgagatgc
     1741 cgagtgccgc gccgcccgcc ggttttttcg atggagtcga gcgtgtgctt ggcaccttca
     1801 aacgtagata cgagaaagag cgtggcccag gatcagtgca attgatgctt aaactcgctc
     1861 caaatgtgtt ctctgatgct tggtctacgg gaggcgcccc ggctgcagtg gctgcagtgg
     1921 aggcttatgt ggagcgcgcg agggccaacg tgtttggtcc tgatggtggt gttacaccgg
     1981 aaggtgtacc agagccactc gacctcgtgc agcttgtctg gtgggacttc gcggcggctg
     2041 atcccttgcc cgttcttaag gctcttcagc gcatggccac agaccagctc caagtggatg
     2101 aagacagcgg tgaggtctct gtatccgaac caaagaaaat tcgtggcatt ggccttgtgg
     2161 acttccccgc cgatcgcctc aaagcggcga tacaagcggg agtccccata acttgcgtcc
     2221 aggtcgagca ctcagtgctt gtaagatccg cccaacctgt actcgatctg tgtgcaaaat
     2281 acggaatcaa ggtactggcc aggggtggaa ctctgggtgg ccttctttcc gcgaaatact
     2341 tgggtgctcc acccccggat ccggttagag gtgatgcgga tctggattct gtacccggct
     2401 gtctggatgc ggttaacaat gtcggtggct gggctcgtct ccaagcggca ctcgcagtaa
     2461 ttaaaggtat agcagacaag catggcgtca aacctgaaac tgttgcattg cgttggcaga
     2521 tcgacgcagg atgttttccg ctcgtaacta cgaggtggtc atcgcgtgta tggcgccagt
     2581 tcggttacga aggttggagt tcgtttgagg tgagtggcgg cagaccagga gtcgatggac
     2641 cactttttca agtcgaaagt ttcctcgacg tagaagatgt acgtgctctt gcgggactgg
     2701 ctgcggttca tctgggtccc aaagcgggct aaaagcttgt cgagaagtac tagaggatca
     2761 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     2821 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     2881 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     2941 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatctgat
     3001 cactgcttga gcctagaaga tccggctgct aacaaagccc gaaaggaagc tgagttggct
     3061 gctgccaccg ctgagcaata actatcataa cccctaggag atccgaacca gataagtgaa
     3121 atctagttcc aaactatttt gtcattttta attttcgtat tagcttacga cgctacaccc
     3181 agttcccatc tattttgtca ctcttcccta aataatcctt aaaaactcca tttccacccc
     3241 tcccagttcc caactatttt gtccgcccac agcggggcat ttttcttcct gttatgtttt
     3301 taatcaaaca tcctgccaac tccatgtgac aaaccgtcat cttcggctac tttttctctg
     3361 tcacagaatg aaaatttttc tgtcatctct tcgttattaa tgtttgtaat tgactgaata
     3421 tcaacgctta tttgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     3481 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3541 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3601 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3661 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3721 cctttgacgt tggagtccac gttcttaata gtggactctt gttccaaact ggaacaacac
     3781 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     3841 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     3901 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3961 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4021 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4081 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4141 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4201 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4261 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4321 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4381 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4441 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4501 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4561 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4621 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4681 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4741 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4801 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4861 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4921 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4981 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5041 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5101 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5161 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactattct
     5221 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5281 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5341 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5401 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5461 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5521 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5581 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5641 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5701 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     5761 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     5821 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     5881 tatttcacac cgcatagacc agccgcgtaa cctggcaaaa tcggttacgg ttgagtaata
     5941 aatggatgcc ctgcgtaagc gggtgtgggc ggacaataaa gtcttaaact gaacaaaata
     6001 gatctaaact atgacaataa agtcttaaac tagacagaat agttgtaaac tgaaatcagt
     6061 ccagttatgc tgtgaaaaag catactggac ttttgttatg gctaaagcaa actcttcatt
     6121 ttctgaagtg caaattgccc gtcgtattaa agaggggcgt ggccaagggc atggtaaaga
     6181 ctatattcgc ggcgttgtga caatttaccg aacaactccg cggccgggaa gccgatctcg
     6241 gcttgaacga attgttaggt ggcggtactt gggtcgatat caaagtgcat cacttcttcc
     6301 cgtatgccca actttgtata gagagccact gcgggatcgt caccgtaatc tgcttgcacg
     6361 tagatcacat aagcaccaag cgcgttggcc tcatgcttga ggagattgat gagcgcggtg
     6421 gcaatgccct gcctccggtg ctcgccggag actgcgagat catagat
//
