LOCUS       sequence-346861-        5924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Chlamydomonas reinhardtii RSP16 recombinant protein in
            insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5924)
  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 5924)
  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-RSP16
FEATURES             Location/Qualifiers
     source          1..5924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(92..97,98..114,115..120))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(516..549)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        597..688
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     613..630
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    1901..2035
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1938..1957)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1992..2011
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2145..2310
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2494..2948
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2581..2600)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2790..2811
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2974..3078
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3079..3147,3148..3939)
                     /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(3297..3316)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4110..4698
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4599..4618
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4852..4869
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  5003..5227
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(5294..5827)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca
       61 gttgcgtgag cgcatacgct acttgcatta cagtttacga accgaacagg cttatgtcaa
      121 ctgggtttgt gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct tgggcagcag
      181 cgaagtcgag gcatttctgt cctggctggc gaacgagcgc aaggtttcgg tctccacgca
      241 tcgtcaggca ttggcggcct tgctgttctt ctacggcaag gtgctgtgca cggatctgcc
      301 cttgcttcag gagatcggta gacctcggcc gtcgcggcgc ttgccggtgg tgctgacccc
      361 ggatgaagtg gttcgcatcc tcggttttct ggaaggcgag catcgtttgt tcgcccagga
      421 ctctagctat agttctagtg gttggctaca gctttgtttg tactatcaac aggttgaact
      481 gctgatcaac agatcctcta cgcggccgcg gtaccataac ttcgtatagc atacattata
      541 cgaagttatc tggttttact agtatcgatt cgcgacctac tccggaatat taatagatca
      601 tggagataat taaaatgata accatctcgc aaataaataa gtattttact gttttcgtaa
      661 cagttttgta ataaaaaaac ctataaatat tccggattat tcataccgtc ccaccatcgg
      721 gcgcggatcc atgacccgcg gccttgatta ctatgaagta atgggattga cccgttctgc
      781 aaacgatata gacattaggc gtgcttacag acgcttggcc cttaagtatc accccgacat
      841 taataaagac ggtgcggctg gagacgaatt ccttcgtatc tgtgaggcct acgaagttct
      901 ctgcgaccca aagacaaagg gtttttatga cctctatggt gaggatgcct tgaaagacgg
      961 tatatcggac ggaaatggtg gcttgaaagg acccatgtat aggttcaacc ctgaagagag
     1021 tccgaaagcc gtatttgagc gtttctttgg cacggcgaac ccctacgagg cgttggaagc
     1081 cttgtcaaat caatttgaaa gcatgacgtc agaggaagcc ccggctcgtg gcaaaaacaa
     1141 agtgtaccct ctcgaattga cactggagga aattttccac ggatgcctca agaaagtggc
     1201 tcataaacgt aaggtcctgt tgttcagcgg cgagtacgtt gaggaggaga gacaactgac
     1261 ggtggacgta aagcccggac tccccacagg aacgaggttc gtatttgagg gtgaaggaaa
     1321 taagactccg aagaaggaac caggtccagt agtattcgtc ctcaagccga agccacatcc
     1381 aagattcgtt cgccgcggtt ccgatctcgt gcataaagtt acccttccgc tgcatcatgc
     1441 cctgatcgga acgacgctcg acatcagaac gctggatgac cgcgacctta aagttccaat
     1501 ctcggatata atgaggcccg gaagctctct cacggtccca ggcgagggta tgccactccc
     1561 ggcgacccct tcagcgcgcg gcaaccttgt gatcgaaata gatctcctct tccccaccca
     1621 tttgaccgaa actcaaaaaa tgttgctgcg ttctgctttc tttctcccgc cacctacaga
     1681 acaaaatgaa gagaccaaga aagcgctgcg tgattacgaa gcagccttta agcatgatct
     1741 caagggatgg gcgacagtgt tcaagcgcta aaagcttgtc gagaagtact agaggatcat
     1801 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     1861 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     1921 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     1981 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatctgatc
     2041 actgcttgag cctagaagat ccggctgcta acaaagcccg aaaggaagct gagttggctg
     2101 ctgccaccgc tgagcaataa ctatcataac ccctaggaga tccgaaccag ataagtgaaa
     2161 tctagttcca aactattttg tcatttttaa ttttcgtatt agcttacgac gctacaccca
     2221 gttcccatct attttgtcac tcttccctaa ataatcctta aaaactccat ttccacccct
     2281 cccagttccc aactattttg tccgcccaca gcggggcatt tttcttcctg ttatgttttt
     2341 aatcaaacat cctgccaact ccatgtgaca aaccgtcatc ttcggctact ttttctctgt
     2401 cacagaatga aaatttttct gtcatctctt cgttattaat gtttgtaatt gactgaatat
     2461 caacgcttat ttgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     2521 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     2581 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     2641 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     2701 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     2761 ctttgacgtt ggagtccacg ttcttaatag tggactcttg ttccaaactg gaacaacact
     2821 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2881 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     2941 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3001 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3061 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     3121 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     3181 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3241 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3301 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     3361 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3421 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3481 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3541 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3601 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     3661 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3721 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3781 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     3841 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     3901 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     3961 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4021 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4081 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4141 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4201 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactattctt
     4261 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4321 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4381 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4441 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4501 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4561 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4621 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4681 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4741 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4801 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     4861 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     4921 atttcacacc gcatagacca gccgcgtaac ctggcaaaat cggttacggt tgagtaataa
     4981 atggatgccc tgcgtaagcg ggtgtgggcg gacaataaag tcttaaactg aacaaaatag
     5041 atctaaacta tgacaataaa gtcttaaact agacagaata gttgtaaact gaaatcagtc
     5101 cagttatgct gtgaaaaagc atactggact tttgttatgg ctaaagcaaa ctcttcattt
     5161 tctgaagtgc aaattgcccg tcgtattaaa gaggggcgtg gccaagggca tggtaaagac
     5221 tatattcgcg gcgttgtgac aatttaccga acaactccgc ggccgggaag ccgatctcgg
     5281 cttgaacgaa ttgttaggtg gcggtacttg ggtcgatatc aaagtgcatc acttcttccc
     5341 gtatgcccaa ctttgtatag agagccactg cgggatcgtc accgtaatct gcttgcacgt
     5401 agatcacata agcaccaagc gcgttggcct catgcttgag gagattgatg agcgcggtgg
     5461 caatgccctg cctccggtgc tcgccggaga ctgcgagatc atagatatag atctcactac
     5521 gcggctgctc aaacttgggc agaacgtaag ccgcgagagc gccaacaacc gcttcttggt
     5581 cgaaggcagc aagcgcgatg aatgtcttac tacggagcaa gttcccgagg taatcggagt
     5641 ccggctgatg ttgggagtag gtggctacgt ctccgaactc acgaccgaaa agatcaagag
     5701 cagcccgcat ggatttgact tggtcagggc cgagcctaca tgtgcgaatg atgcccatac
     5761 ttgagccacc taactttgtt ttagggcgac tgccctgctg cgtaacatcg ttgctgctgc
     5821 gtaacatcgt tgctgctcca taacatcaaa catcgaccca cggcgtaacg cgcttgctgc
     5881 ttggatgccc gaggcataga ctgtacaaaa aaacagtcat aaca
//
