LOCUS       sequence-346859-        6386 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Chlamydomonas reinhardtii RSP7 recombinant protein in 
            insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6386)
  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 6386)
  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-RSP7
FEATURES             Location/Qualifiers
     source          1..6386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(82..87,88..104,105..110))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(506..539)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        587..678
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     603..620
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    2353..2487
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2390..2409)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2444..2463
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2597..2762
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2946..3400
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3033..3052)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3242..3263
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3426..3530
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3531..3599,3600..4391)
                     /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(3749..3768)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4562..5150
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5051..5070
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5304..5321
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  5455..5679
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(5746..6279)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca gttgcgtgag
       61 cgcatacgct acttgcatta cagtttacga accgaacagg cttatgtcaa ctgggtttgt
      121 gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct tgggcagcag cgaagtcgag
      181 gcatttctgt cctggctggc gaacgagcgc aaggtttcgg tctccacgca tcgtcaggca
      241 ttggcggcct tgctgttctt ctacggcaag gtgctgtgca cggatctgcc cttgcttcag
      301 gagatcggta gacctcggcc gtcgcggcgc ttgccggtgg tgctgacccc ggatgaagtg
      361 gttcgcatcc tcggttttct ggaaggcgag catcgtttgt tcgcccagga ctctagctat
      421 agttctagtg gttggctaca gctttgtttg tactatcaac aggttgaact gctgatcaac
      481 agatcctcta cgcggccgcg gtaccataac ttcgtatagc atacattata cgaagttatc
      541 tggttttact agtatcgatt cgcgacctac tccggaatat taatagatca tggagataat
      601 taaaatgata accatctcgc aaataaataa gtattttact gttttcgtaa cagttttgta
      661 ataaaaaaac ctataaatat tccggattat tcataccgtc ccaccatcgg gcgcggatcc
      721 atgagtagca cttaccagaa gcctataacc atcccaggtg actttccggc tatactcaag
      781 gcgttcacga gagagatatt gcgcgcccag ccatccaaca tttatgaatt tggagcacgt
      841 tattttagtg gcttgcaagg acaggccgaa ccggagggcc tcggtagggc ggcaccggtg
      901 gaaccggcgt ctacaagcca cgcagcaaca agcaaggcca ctgatgtaga ggagacggcg
      961 gtgatgtttg atattgccgc acttacccca gcggaacttg agcccatact tatgaaactc
     1021 ttcatcgaag cagacgcgga tcgttcgggc tttttggatc gtcatgagtt tactgctgtg
     1081 ttgcgcaacg ctaatctcaa gctttcggat aggcagatac gccagatcct tgccgaggca
     1141 gatgaaaatg atgacgacgt gatacaatac aaagaattcc tcccgatcat ggttgatata
     1201 ttgcaaagca tcaaggcaaa agagcaagcg aaggcaatga tgcatggagt cgaaaccatg
     1261 gtgcgcactg aagtagagac gatgctcctc cacggtttgc cccaggaaga gcttcaggcc
     1321 ttgatgctca aagtattcaa aaaagcggac gcggacggct ctggacagtt gaaccgccac
     1381 gaattcaaag aggccctgaa ggcagccgag ttgggcctca caccaaaaga tattaatttg
     1441 atactgtctc acattgacgt tgacagagat ggtcttgtca gctatgagga attcattcca
     1501 gtgtgtttcc aagttctcgt agaaagattc aaggatgaga tagtcgttaa tgacattctt
     1561 ggcaacgctg atgaattgca gcagatgttg ttgggcgcct tccgcgatgc tgatccggac
     1621 aatacgggcc tcttgagcca acgccaagta aagagcatat tcaaggagtt gtcatataaa
     1681 gctttgggac tgactacgtt gcagatggtt tcactcatta gccaggcacc aacgacccca
     1741 gacggaatgg tgcagtacat acaatttgtg cctcaggcgg cgagcataat ccgtagtatg
     1801 tatgacgtag agactatgaa aggccgtatg catgcaatta aggcggtcgc cgaagcgggt
     1861 ggaatagctg ccctcggtgc cttggatctc gaccagctgc gcggtgttct tgagcaagcc
     1921 tttcagagag tggatactga aggcgctggt cagttgactt tgccgcaagt gacacaagtg
     1981 ctcgacggcc ttaactcctt ggctcctgac gccaacttgg ccctctcaga ccagcatatg
     2041 aaggcgatgt ttgcggcgat agacgcagac gaaagcggca cggtagactg gacggaattg
     2101 gttaatttta tatgtgacgc gctggagcac atagagaggg aagcgtatgt tgcaaatatg
     2161 cgtgacggcg gagcaggtgg agcgggcgaa gcggaggctt cccccggcga cgaagaggcg
     2221 taaaagcttg tcgagaagta ctagaggatc ataatcagcc ataccacatt tgtagaggtt
     2281 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     2341 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     2401 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2461 atcaatgtat cttatcatgt ctggatctga tcactgcttg agcctagaag atccggctgc
     2521 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactatcata
     2581 acccctagga gatccgaacc agataagtga aatctagttc caaactattt tgtcattttt
     2641 aattttcgta ttagcttacg acgctacacc cagttcccat ctattttgtc actcttccct
     2701 aaataatcct taaaaactcc atttccaccc ctcccagttc ccaactattt tgtccgccca
     2761 cagcggggca tttttcttcc tgttatgttt ttaatcaaac atcctgccaa ctccatgtga
     2821 caaaccgtca tcttcggcta ctttttctct gtcacagaat gaaaattttt ctgtcatctc
     2881 ttcgttatta atgtttgtaa ttgactgaat atcaacgctt atttgcagcc tgaatggcga
     2941 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3001 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3061 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3121 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3181 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttcttaat
     3241 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3301 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3361 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     3421 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3481 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     3541 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3601 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3661 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     3721 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     3781 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3841 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3901 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3961 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4021 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4081 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4141 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4201 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     4261 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4321 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4381 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4441 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4501 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4561 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4621 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4681 tcagcagagc gcagatacca aatactattc ttctagtgta gccgtagtta ggccaccact
     4741 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4801 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4861 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4921 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4981 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5041 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5101 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5161 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     5221 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     5281 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     5341 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatagac cagccgcgta
     5401 acctggcaaa atcggttacg gttgagtaat aaatggatgc cctgcgtaag cgggtgtggg
     5461 cggacaataa agtcttaaac tgaacaaaat agatctaaac tatgacaata aagtcttaaa
     5521 ctagacagaa tagttgtaaa ctgaaatcag tccagttatg ctgtgaaaaa gcatactgga
     5581 cttttgttat ggctaaagca aactcttcat tttctgaagt gcaaattgcc cgtcgtatta
     5641 aagaggggcg tggccaaggg catggtaaag actatattcg cggcgttgtg acaatttacc
     5701 gaacaactcc gcggccggga agccgatctc ggcttgaacg aattgttagg tggcggtact
     5761 tgggtcgata tcaaagtgca tcacttcttc ccgtatgccc aactttgtat agagagccac
     5821 tgcgggatcg tcaccgtaat ctgcttgcac gtagatcaca taagcaccaa gcgcgttggc
     5881 ctcatgcttg aggagattga tgagcgcggt ggcaatgccc tgcctccggt gctcgccgga
     5941 gactgcgaga tcatagatat agatctcact acgcggctgc tcaaacttgg gcagaacgta
     6001 agccgcgaga gcgccaacaa ccgcttcttg gtcgaaggca gcaagcgcga tgaatgtctt
     6061 actacggagc aagttcccga ggtaatcgga gtccggctga tgttgggagt aggtggctac
     6121 gtctccgaac tcacgaccga aaagatcaag agcagcccgc atggatttga cttggtcagg
     6181 gccgagccta catgtgcgaa tgatgcccat acttgagcca cctaactttg ttttagggcg
     6241 actgccctgc tgcgtaacat cgttgctgct gcgtaacatc gttgctgctc cataacatca
     6301 aacatcgacc cacggcgtaa cgcgcttgct gcttggatgc ccgaggcata gactgtacaa
     6361 aaaaacagtc ataacaagcc atgaaa
//
