LOCUS       sequence-346870-        7328 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Chlamydomonas reinhardtii RSP1 recombinant protein in 
            insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7328)
  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 7328)
  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-RSP1
FEATURES             Location/Qualifiers
     source          1..7328
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(110..115,116..132,133..138))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(534..567)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        615..706
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     631..648
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    3323..3457
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3360..3379)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3414..3433
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3567..3732
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3916..4370
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4003..4022)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4212..4233
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4396..4500
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4501..4569,4570..5361)
                     /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(4719..4738)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5532..6120
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6021..6040
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6274..6291
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6425..6649
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6716..7249)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 aaaaaaacag tcataacaag ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc
       61 ggtcaaggtt ctggaccagt tgcgtgagcg catacgctac ttgcattaca gtttacgaac
      121 cgaacaggct tatgtcaact gggtttgtgc cttcatccgt ttccacggtg tgcgtcaccc
      181 ggcaaccttg ggcagcagcg aagtcgaggc atttctgtcc tggctggcga acgagcgcaa
      241 ggtttcggtc tccacgcatc gtcaggcatt ggcggccttg ctgttcttct acggcaaggt
      301 gctgtgcacg gatctgccct tgcttcagga gatcggtaga cctcggccgt cgcggcgctt
      361 gccggtggtg ctgaccccgg atgaagtggt tcgcatcctc ggttttctgg aaggcgagca
      421 tcgtttgttc gcccaggact ctagctatag ttctagtggt tggctacagc tttgtttgta
      481 ctatcaacag gttgaactgc tgatcaacag atcctctacg cggccgcggt accataactt
      541 cgtatagcat acattatacg aagttatctg gttttactag tatcgattcg cgacctactc
      601 cggaatatta atagatcatg gagataatta aaatgataac catctcgcaa ataaataagt
      661 attttactgt tttcgtaaca gttttgtaat aaaaaaacct ataaatattc cggattattc
      721 ataccgtccc accatcgggc gcggatccat ggcccttgga cccttcgtcc tcccatttcg
      781 cggtgaccag tatagctttg gtattaattt taaaagctca ccagaagaaa agctgaactt
      841 cgatttgtca tgtgtagctt ttgacgttaa gggtcagctg cacgacacgc tccatgctcg
      901 caagccgacc gcattggacg gcgccttggt taaaggtttc gaaaaacaag cattgccgga
      961 ggaaacagta caagtcgagg gtgatgacgt gatctatatg ttcccgaaaa agtttgagag
     1021 acaagttgaa gtgctgctgt ttgttgccag tgccccaagt atccccggca agaagcacga
     1081 cctggattcc tcaagcaagc tcgagtttgc ggtgagttat agcgacgtcg gaggacaagc
     1141 ctttaaccag tcatttgatc tgaaaccact ggcagcgcaa ggaggcgtat caagcataat
     1201 agttgccgta atgtacttgc aggccgaagg tggatggacg cttcgcagtg taggcgattg
     1261 tcacccattt gacagccccg gtttgatagt gccggaactt aaacaaacca ttctcaacct
     1321 gcgcgatcat cacggcgtcc agttggacgc ggctgacgca atccaagcta tagaccccgc
     1381 tgagcgcgtt ccagtcaccc gccaattcca ggaccagtct ttggacgaag cgtctgcggg
     1441 acgtgcagcc gaacctgcac ctgttaagaa attgaggata gatttgtcct ggaccttctg
     1501 gccgcctcca cctccaacgg aagagggtga agagccccca gaagaaccgg ctcttgaata
     1561 taaccttgtt atgtataaca aggacggaga ggaagtacag tctatctcca ccggtaaccg
     1621 tgaggctaca ggtgcccgtg ccggccgtcc ggaaccggag gaggacgaag aagaggaaaa
     1681 ggaggaggag aaagaggagc cggaggaggg agaagaggga gaagaaggtg agggcggaga
     1741 gccgaaggag cctcctccgc cccctccggc tcccaaagtc gatccgtatg aatttaagga
     1801 gcgcgacgta atctatcttg acgtaccaga ccttcctgct gaggtccgca gtatggttct
     1861 cctggtaacc aactacgatg aagagaatgg cttcactcgc gtacgtacag tgcgctgccg
     1921 ccttgttgac gtcagcaacg gagaggcccc cctgccaggt tcaaaggccg cggtagcagc
     1981 ggctgctgct gctgctgaac aaggtctggc agccccccct aacccagaga gagtacttgc
     2041 cgactacggc gtcctctcaa agtatgaaga tgataaggca actacccaag tcgccctcat
     2101 gaagttgtac aaagaatatg cagattcggc atttaacgta ttcagaggag caggagttga
     2161 taacgtggct gcctttatcg gacaagagcc ggacactatt atcaatcagc ttaaagccta
     2221 tctggaagcg actaaaaaac agaaggcagc tgaggcagca gcagccgctg ccgcggagga
     2281 atccggagaa gagattacag cagatcccaa gccgcatgtc tggagatttc gcgcgctcgg
     2341 tttgaacttc ggtggagact cacttgaagc aatagagcat gatttgaaga atctttttgc
     2401 ctttgatgga gacctcgccc ccggagctgc aagggattca gatacctcaa ggtcaagctt
     2461 cccgaatggt gatacttatt tcggcagcta cgcggacgat gtgaagcatg gcccgggact
     2521 ctatgcgttt gcgacgggcg ccggatacgc tggcgagtac gcaggcggaa aacgtcatgg
     2581 ccgtggagtc atggtattcc cagacggcgg tacctacgta ggtgagtttg tcgctgataa
     2641 attcgagggt cagggacagt atcgctatcc cgacggcagt gtctacaccg gcagttgggc
     2701 cgcgggacag aagcacggtc caggcgttta ttgggatact gcgaggggat gtctgagggg
     2761 cgaatggaaa aagggattgc tggtcggaaa gggcacatac gaacagcccg cgcttcgttt
     2821 tgaaggcgaa ttcgttagag gtatgcccgc tggcaccgcg acttatactc tgactggtca
     2881 ccgtactttg gatatgcctt gttttgcggc acaacatatc caggctgagg aaggacccac
     2941 tttggccctt ccgtgcgcct atggcatacc gcccggctca ggagatgagc cgcagcttga
     3001 tgaagagggt caacctatag aggatactga taaacctccg cttccggcac acccaaagta
     3061 tgagggtctt accttcactg ctgagcaatt gcctggcgcg gcaccagaca ctgtgtttcc
     3121 acctgaagag ggtaaacctg ttccaatcac ggctgtgcct gctttctcgg tctccacagg
     3181 attggtggct taaaagcttg tcgagaagta ctagaggatc ataatcagcc ataccacatt
     3241 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     3301 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     3361 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3421 gtccaaactc atcaatgtat cttatcatgt ctggatctga tcactgcttg agcctagaag
     3481 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     3541 aactatcata acccctagga gatccgaacc agataagtga aatctagttc caaactattt
     3601 tgtcattttt aattttcgta ttagcttacg acgctacacc cagttcccat ctattttgtc
     3661 actcttccct aaataatcct taaaaactcc atttccaccc ctcccagttc ccaactattt
     3721 tgtccgccca cagcggggca tttttcttcc tgttatgttt ttaatcaaac atcctgccaa
     3781 ctccatgtga caaaccgtca tcttcggcta ctttttctct gtcacagaat gaaaattttt
     3841 ctgtcatctc ttcgttatta atgtttgtaa ttgactgaat atcaacgctt atttgcagcc
     3901 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3961 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     4021 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     4081 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     4141 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     4201 cgttcttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4261 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4321 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     4381 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4441 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4501 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4561 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     4621 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     4681 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     4741 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     4801 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     4861 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     4921 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     4981 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5041 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5101 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5161 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5221 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     5281 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     5341 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     5401 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5461 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5521 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5581 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5641 gtaactggct tcagcagagc gcagatacca aatactattc ttctagtgta gccgtagtta
     5701 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5761 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5821 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5881 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5941 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6001 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6061 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6121 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6181 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     6241 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     6301 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatagac
     6361 cagccgcgta acctggcaaa atcggttacg gttgagtaat aaatggatgc cctgcgtaag
     6421 cgggtgtggg cggacaataa agtcttaaac tgaacaaaat agatctaaac tatgacaata
     6481 aagtcttaaa ctagacagaa tagttgtaaa ctgaaatcag tccagttatg ctgtgaaaaa
     6541 gcatactgga cttttgttat ggctaaagca aactcttcat tttctgaagt gcaaattgcc
     6601 cgtcgtatta aagaggggcg tggccaaggg catggtaaag actatattcg cggcgttgtg
     6661 acaatttacc gaacaactcc gcggccggga agccgatctc ggcttgaacg aattgttagg
     6721 tggcggtact tgggtcgata tcaaagtgca tcacttcttc ccgtatgccc aactttgtat
     6781 agagagccac tgcgggatcg tcaccgtaat ctgcttgcac gtagatcaca taagcaccaa
     6841 gcgcgttggc ctcatgcttg aggagattga tgagcgcggt ggcaatgccc tgcctccggt
     6901 gctcgccgga gactgcgaga tcatagatat agatctcact acgcggctgc tcaaacttgg
     6961 gcagaacgta agccgcgaga gcgccaacaa ccgcttcttg gtcgaaggca gcaagcgcga
     7021 tgaatgtctt actacggagc aagttcccga ggtaatcgga gtccggctga tgttgggagt
     7081 aggtggctac gtctccgaac tcacgaccga aaagatcaag agcagcccgc atggatttga
     7141 cttggtcagg gccgagccta catgtgcgaa tgatgcccat acttgagcca cctaactttg
     7201 ttttagggcg actgccctgc tgcgtaacat cgttgctgct gcgtaacatc gttgctgctc
     7261 cataacatca aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata
     7321 gactgtac
//
