LOCUS       sequence-346974-        7100 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Chlamydomonas reinhardtii RSP2 recombinant protein in 
            insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7100)
  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 7100)
  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-RSP2
FEATURES             Location/Qualifiers
     source          1..7100
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(1..6,7..23,24..29))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(425..458)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        506..597
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     522..539
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    2986..3120
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3023..3042)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3077..3096
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3230..3395
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3579..4033
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3666..3685)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3875..3896
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4059..4163
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4164..4232,4233..5024)
                     /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(4382..4401)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5195..5783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5684..5703
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5937..5954
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6088..6312
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6379..6912)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 agtttacgaa ccgaacaggc ttatgtcaac tgggtttgtg ccttcatccg tttccacggt
       61 gtgcgtcacc cggcaacctt gggcagcagc gaagtcgagg catttctgtc ctggctggcg
      121 aacgagcgca aggtttcggt ctccacgcat cgtcaggcat tggcggcctt gctgttcttc
      181 tacggcaagg tgctgtgcac ggatctgccc ttgcttcagg agatcggtag acctcggccg
      241 tcgcggcgct tgccggtggt gctgaccccg gatgaagtgg ttcgcatcct cggttttctg
      301 gaaggcgagc atcgtttgtt cgcccaggac tctagctata gttctagtgg ttggctacag
      361 ctttgtttgt actatcaaca ggttgaactg ctgatcaaca gatcctctac gcggccgcgg
      421 taccataact tcgtatagca tacattatac gaagttatct ggttttacta gtatcgattc
      481 gcgacctact ccggaatatt aatagatcat ggagataatt aaaatgataa ccatctcgca
      541 aataaataag tattttactg ttttcgtaac agttttgtaa taaaaaaacc tataaatatt
      601 ccggattatt cataccgtcc caccatcggg cgcggatcca tggctcccac ccaagctggt
      661 cacgacactg cctatttaaa ggaaaccgtg ggtgaggctc tggctcgtgg ttgcgctgct
      721 gctatcagcg cccaacccaa cgatcccgtt gagtacctcg gtctgtggtt attaaagtac
      781 gtcaagaacg ctgaggtgga gggcaatttc tatcgcgagc gccagcaaga tttacaaaag
      841 aagaaggacc gtttagtcaa ggaagcccag tctgagcaag ctgctaagag cgtggctctg
      901 actcgcaagg aggctgccga cgctctcgct ttagtgactg ccgagcctcg tgagctgctg
      961 gaagctgccg tcaagctcgt gaagcagcac accgctgctg gtgccgccta tgccgccgtc
     1021 gtggctgaac ccgaggagcc cgactgggtc gctcccgaag atgacgaagc tgctgctgtg
     1081 gagaccgagg acgaagccgc tggtggtgct gctttagctg aaggcgagga acctcccccc
     1141 gaacccgaac ccgaacccga agctgccccc gaagatggcg agggtgatgc ccccgctcct
     1201 aagatccctc gccccgtgga ttacagcaag aagtacttcg cttacgtcgc tgctagcgcc
     1261 ggtcaagaac atgtgctcga ggctgattta taccgccccg ctcctcctcc cgaagacgct
     1321 gatgaggact tcaagcccga acctttaccc tactccttcc gcgttttaga cgagaaactc
     1381 cctatgctgt acgtgcccaa cgttgccgct gaagagcgcg tcaagttctt tcgtaagttc
     1441 cccaagatcg gctcctacca agcttgtggt gtcgccctcc ccgctagcgg tgagttcaag
     1501 gctctgctgg ctgctgatac tttattcccc gaaggttccg gccaacctct gtccgccgat
     1561 gaccgtgatt tcgtctggga ggtctcccag tctttaagcc gtgctttaga agctgtgcaa
     1621 gctcgcgccg ctgaggcttt agctgctacc tccgctgccg aagctgtgga agaactgaag
     1681 gccaaggtgg ctgagctgcg cgagcaagct gctgccgagg ctgccgctgc tgctcccccc
     1741 ccccccgctg aaggtgaaga gggtgagggt gaagcccccc ccgctgaaga agagcctccc
     1801 gctgaagaag aagctgaaga ggaggaagag gaggccgagg aaggtgctga ggagggtgcc
     1861 gaagaaggtg aggagggcga ggaagctccc cccaagccta aaaagaagaa gaaggtgttc
     1921 aaccctatcc ccggtttaca agctgctatc gagaagctca ccgctgccgc tgaagccgct
     1981 actgaggctg acgctcgtgc tcaagctgct gtcgctctgg agaagcaagc tttagacgag
     2041 gtggtggctc tggcctccag ccatagcgac gctactctgt cctctttacg caatatgctc
     2101 agcgtgcctc aaggtaccta ccacgtcgtc aaggctttat tacacttatt aggccgtccc
     2161 gctgctagct tctccacttg gaagcgtgct cactcccact tcagccctcg tctgttcgag
     2221 gacatggctg cctacgatgc tactgccgaa cgtgacatgg ctgtgtgggg ccgcgtgcgc
     2281 tcttgttaca aggccgctcc cgccgccaag aagctggacg ccgagatgcc caacactctg
     2341 ttcggttccg tggccctcat gtacatcaaa caagttcgcc gcgtggctcg caaagctgtg
     2401 ctccaacgcg aactggccgc caagctcgct aaggcccaac aagatttagc cgacaagcaa
     2461 gctgctctgg tcgaggctga acgcgtgaag gctgagcgcg aggctgagga agcccgtttg
     2521 gccgccgaag ccgaagctgc cgccgctgct gaagctgaag ctgctgctcg cgccgccgcc
     2581 gaagctgagg ctgctgccgc tgctgaagct gccgctgaag ctgccgctga ggccgccgcc
     2641 gccgctgccg aggctgctgc tgaggctggc gaaggtgagg ctgtggccga acgcgaagct
     2701 gctcccgctg aggccgaggc tgcccccgcc gaaggcgagg ctgctcctcc cgctgaaggc
     2761 gaaggcgaag ctcaacccgc tcaagaaggt tccaactcct cctccagctc cagcgacagc
     2821 tcctccagcg aggaatccaa agctgctgcc gagtaaaagc ttgtcgagaa gtactagagg
     2881 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     2941 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3001 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3061 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     3121 tgatcactgc ttgagcctag aagatccggc tgctaacaaa gcccgaaagg aagctgagtt
     3181 ggctgctgcc accgctgagc aataactatc ataaccccta ggagatccga accagataag
     3241 tgaaatctag ttccaaacta ttttgtcatt tttaattttc gtattagctt acgacgctac
     3301 acccagttcc catctatttt gtcactcttc cctaaataat ccttaaaaac tccatttcca
     3361 cccctcccag ttcccaacta ttttgtccgc ccacagcggg gcatttttct tcctgttatg
     3421 tttttaatca aacatcctgc caactccatg tgacaaaccg tcatcttcgg ctactttttc
     3481 tctgtcacag aatgaaaatt tttctgtcat ctcttcgtta ttaatgtttg taattgactg
     3541 aatatcaacg cttatttgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt
     3601 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3661 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3721 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3781 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3841 tcgccctttg acgttggagt ccacgttctt aatagtggac tcttgttcca aactggaaca
     3901 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3961 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     4021 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4081 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4141 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4201 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4261 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     4321 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     4381 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4441 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     4501 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4561 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     4621 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4681 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     4741 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     4801 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4861 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4921 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4981 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5041 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5101 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5161 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5221 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5281 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatacta
     5341 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5401 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5461 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5521 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5581 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5641 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5701 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5761 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5821 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     5881 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     5941 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     6001 gcggtatttc acaccgcata gaccagccgc gtaacctggc aaaatcggtt acggttgagt
     6061 aataaatgga tgccctgcgt aagcgggtgt gggcggacaa taaagtctta aactgaacaa
     6121 aatagatcta aactatgaca ataaagtctt aaactagaca gaatagttgt aaactgaaat
     6181 cagtccagtt atgctgtgaa aaagcatact ggacttttgt tatggctaaa gcaaactctt
     6241 cattttctga agtgcaaatt gcccgtcgta ttaaagaggg gcgtggccaa gggcatggta
     6301 aagactatat tcgcggcgtt gtgacaattt accgaacaac tccgcggccg ggaagccgat
     6361 ctcggcttga acgaattgtt aggtggcggt acttgggtcg atatcaaagt gcatcacttc
     6421 ttcccgtatg cccaactttg tatagagagc cactgcggga tcgtcaccgt aatctgcttg
     6481 cacgtagatc acataagcac caagcgcgtt ggcctcatgc ttgaggagat tgatgagcgc
     6541 ggtggcaatg ccctgcctcc ggtgctcgcc ggagactgcg agatcataga tatagatctc
     6601 actacgcggc tgctcaaact tgggcagaac gtaagccgcg agagcgccaa caaccgcttc
     6661 ttggtcgaag gcagcaagcg cgatgaatgt cttactacgg agcaagttcc cgaggtaatc
     6721 ggagtccggc tgatgttggg agtaggtggc tacgtctccg aactcacgac cgaaaagatc
     6781 aagagcagcc cgcatggatt tgacttggtc agggccgagc ctacatgtgc gaatgatgcc
     6841 catacttgag ccacctaact ttgttttagg gcgactgccc tgctgcgtaa catcgttgct
     6901 gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg taacgcgctt
     6961 gctgcttgga tgcccgaggc atagactgta caaaaaaaca gtcataacaa gccatgaaaa
     7021 ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc
     7081 gcatacgcta cttgcattac
//
