LOCUS       sequence-346863-        6644 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Chlamydomonas reinhardtii RSP23 recombinant protein in
            insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6644)
  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 6644)
  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-RSP23
FEATURES             Location/Qualifiers
     source          1..6644
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(404..437)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        485..576
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     501..518
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    2509..2643
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2546..2565)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2600..2619
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2753..2918
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3102..3556
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3189..3208)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3398..3419
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3582..3686
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3687..3755,3756..4547)
                     /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(3905..3924)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4718..5306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5207..5226
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5460..5477
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  5611..5835
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(5902..6435)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(6624..6629,6630..6644,1..2,3..8))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 tatgtcaact gggtttgtgc cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg
       61 ggcagcagcg aagtcgaggc atttctgtcc tggctggcga acgagcgcaa ggtttcggtc
      121 tccacgcatc gtcaggcatt ggcggccttg ctgttcttct acggcaaggt gctgtgcacg
      181 gatctgccct tgcttcagga gatcggtaga cctcggccgt cgcggcgctt gccggtggtg
      241 ctgaccccgg atgaagtggt tcgcatcctc ggttttctgg aaggcgagca tcgtttgttc
      301 gcccaggact ctagctatag ttctagtggt tggctacagc tttgtttgta ctatcaacag
      361 gttgaactgc tgatcaacag atcctctacg cggccgcggt accataactt cgtatagcat
      421 acattatacg aagttatctg gttttactag tatcgattcg cgacctactc cggaatatta
      481 atagatcatg gagataatta aaatgataac catctcgcaa ataaataagt attttactgt
      541 tttcgtaaca gttttgtaat aaaaaaacct ataaatattc cggattattc ataccgtccc
      601 accatcgggc gcggatccat ggcggagctg gagaagactt ttgcactcat aaagccagat
      661 gctgtccgcg caggcaaggc tcaggaaatc atgcagctta tagagttgaa cggtttcacc
      721 atcattgcta agcaaaagtt gcaactcacg agagcccgcg ctgaggaatt ctatggagag
      781 cacaagggta aggagttttt cccaaagctt gtcaatttta tgacatcagg tccgatttgg
      841 gccttggtgc tggccaaacc tggagcaatt ctcgcttggc gcgcgcttat gggacctacc
      901 aatgtgttca aggcaagggc cgaacaacca aaatgcctcc gcgcactcta cggaacagac
      961 ggaacacaaa atgcaaccca cggcagtgat tcccctatat cagcagcccg cgaaattaag
     1021 ttcttcttcc cgacgctctc gggtgatccc actatttacg cggagcccac cgccgcggca
     1081 gaatatatca cgaaacgcat tcaaccagct ctggcaaagg ctcttgccgc cctggcgcgc
     1141 gagaagccga gcgctgataa atttgaggcc atcacctttg tagcgggcta cctgctccaa
     1201 aataacccca acaagccgaa ggtgcttatg ccggacgagt gggaccccgc gcttatgggt
     1261 gatgatgagg acgacgaagc agatttcatc aacgccagac tcgccgcgcc gacaggaaat
     1321 gacggtgcaa caaaagcaga atttgatgca atggttgaag ctgcaaaagc tgatacagga
     1381 ggtgcagctc cagcccagga atttgtttat cctgattcta aaccaactac tccggttgta
     1441 ccggctcctc cggccccagg ttctaaacct ccatctgcaa gcggtgcgag gccacaaagc
     1501 gcaaggccca cgagtgcaag accgccatcg gcgagcgcag ccccacctgc cccgctggct
     1561 cctgtaccac cgccggcttc ctcaagcaga cctgcgtcgg gtagcggtag gcccccttcg
     1621 gcgactgcgc gccccccatc agctacacct ccaccgcctc cacctgcagt ggagctggaa
     1681 gaggcggatg acccggcgca gctcgacgag gctgcgacaa aggtccaagc tgccttccgt
     1741 ggctaccaag ctaggaagga ggtcgctgtc atgcgctctg aagcccaacc gggagaagca
     1801 gcagccgaac ctgaacaaga ggccgagctg caaccggaag cggagccgga accccagccc
     1861 gagggagagc aagagccgca acctcaagcc tccgcttcat cgtcattcct tccggacggt
     1921 gtgacggagg agatggctgc cgaggcagca acgcgcgtac aggctcatat gaggggacac
     1981 ctggctagga aacaggttgc tgcgattaaa gcgcagcagg cggctccagc agtggcggag
     2041 tcttccgagg ctctcgcaga gcccgaaccg cagcctgaag ctgaagctga accacaacca
     2101 caagcttcag tatcttcttc ttttctgcca gacggagtaa cagaagagat ggcagcagag
     2161 gctgcgacct tggttcaagc tcatatgcgt ggtcacctgg cgcgtaaaca ggttgcagca
     2221 atcaaagctc agcaagccgc ccccgcagtt gccgaaagtt cagaagctca ggccgaagca
     2281 gaacaaacgg aggctgaagc cgagccccaa cctgatgctg aggctgaagc aggagctgct
     2341 gaaggagagg ccgaaccaga gccagaagct gcagaataaa agcttgtcga gaagtactag
     2401 aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     2461 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     2521 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2581 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     2641 atctgatcac tgcttgagcc tagaagatcc ggctgctaac aaagcccgaa aggaagctga
     2701 gttggctgct gccaccgctg agcaataact atcataaccc ctaggagatc cgaaccagat
     2761 aagtgaaatc tagttccaaa ctattttgtc atttttaatt ttcgtattag cttacgacgc
     2821 tacacccagt tcccatctat tttgtcactc ttccctaaat aatccttaaa aactccattt
     2881 ccacccctcc cagttcccaa ctattttgtc cgcccacagc ggggcatttt tcttcctgtt
     2941 atgtttttaa tcaaacatcc tgccaactcc atgtgacaaa ccgtcatctt cggctacttt
     3001 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
     3061 ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
     3121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3241 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3361 ttttcgccct ttgacgttgg agtccacgtt cttaatagtg gactcttgtt ccaaactgga
     3421 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3481 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     3541 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3601 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3661 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3721 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3781 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3841 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3901 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3961 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4021 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4081 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     4141 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4201 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     4261 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     4321 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4381 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4441 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4501 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4561 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4621 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4681 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4741 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4801 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4861 ctattcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4921 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4981 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5041 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5101 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5161 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5221 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5281 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5341 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5401 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     5461 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     5521 tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct ggcaaaatcg gttacggttg
     5581 agtaataaat ggatgccctg cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa
     5641 caaaatagat ctaaactatg acaataaagt cttaaactag acagaatagt tgtaaactga
     5701 aatcagtcca gttatgctgt gaaaaagcat actggacttt tgttatggct aaagcaaact
     5761 cttcattttc tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggc caagggcatg
     5821 gtaaagacta tattcgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc
     5881 gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac
     5941 ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc
     6001 ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttgagga gattgatgag
     6061 cgcggtggca atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat
     6121 ctcactacgc ggctgctcaa acttgggcag aacgtaagcc gcgagagcgc caacaaccgc
     6181 ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta
     6241 atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag
     6301 atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat
     6361 gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt
     6421 gctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg
     6481 cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga
     6541 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
     6601 agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggct
//
