LOCUS       sequence-346977-        5448 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Chlamydomonas reinhardtii RSP12 recombinant protein in
            insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5448)
  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 5448)
  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-RSP12
FEATURES             Location/Qualifiers
     source          1..5448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(494..499,500..516,517..522))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(918..951)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        999..1090
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1015..1032
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    1808..1942
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1845..1864)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1899..1918
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2052..2217
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2401..2855
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2488..2507)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2697..2718
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2881..2985
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2986..3054,3055..3846)
                     /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(3204..3223)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4017..4605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4506..4525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4759..4776
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  4910..5134
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(join(5201..5448,1..286))
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 gaacgtaagc cgcgagagcg ccaacaaccg cttcttggtc gaaggcagca agcgcgatga
       61 atgtcttact acggagcaag ttcccgaggt aatcggagtc cggctgatgt tgggagtagg
      121 tggctacgtc tccgaactca cgaccgaaaa gatcaagagc agcccgcatg gatttgactt
      181 ggtcagggcc gagcctacat gtgcgaatga tgcccatact tgagccacct aactttgttt
      241 tagggcgact gccctgctgc gtaacatcgt tgctgctgcg taacatcgtt gctgctgcgt
      301 aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt
      361 ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga aaaccgccac
      421 tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg agcgcatacg
      481 ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttt gtgccttcat
      541 ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg aggcatttct
      601 gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg cattggcggc
      661 cttgctgttc ttctacggca aggtgctgtg cacggatctg cccttgcttc aggagatcgg
      721 tagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag tggttcgcat
      781 cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct atagttctag
      841 tggttggcta cagctttgtt tgtactatca acaggttgaa ctgctgatca acagatcctc
      901 tacgcggccg cggtaccata acttcgtata gcatacatta tacgaagtta tctggtttta
      961 ctagtatcga ttcgcgacct actccggaat attaatagat catggagata attaaaatga
     1021 taaccatctc gcaaataaat aagtatttta ctgttttcgt aacagttttg taataaaaaa
     1081 acctataaat attccggatt attcataccg tcccaccatc gggcgcggat ccatggattt
     1141 tgaaagcaac ggagctatga tggagtattg caagtcaaca ggtcgcacgt attgtgcatt
     1201 cagtatccaa caatcatcta agcttcttgg tacagttgtc ttggagcttt tcactgatat
     1261 tgcgcccgca acgtgcgcga acttcatcaa atacatcaag gatggatatc agggaactcc
     1321 acttcatcgc attgttccca acggttgggt tcaaggcggc gacattgtcg atggtagcgg
     1381 aaagggagat cctggattcg tgcttccaga cgaaacttat tctgtcaaac atgacgctcc
     1441 gggcgtactt ggcatggcaa caggcggtca gcctcacaca gcaaacaccc agttctatat
     1501 ttccttgtct cctcttccct ttctcgatgg caaaagagtg gccttcggta gggtcctgaa
     1561 caaacaaagt ctcgagaatc ttctcgctct ccaaacactc ccaacgtttc aaaatgaaag
     1621 acctgtcccg gacgtagtta tagcttcatg tcacgtaata tataatccca attcctaaaa
     1681 gcttgtcgag aagtactaga ggatcataat cagccatacc acatttgtag aggttttact
     1741 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     1801 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     1861 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     1921 tgtatcttat catgtctgga tctgatcact gcttgagcct agaagatccg gctgctaaca
     1981 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta tcataacccc
     2041 taggagatcc gaaccagata agtgaaatct agttccaaac tattttgtca tttttaattt
     2101 tcgtattagc ttacgacgct acacccagtt cccatctatt ttgtcactct tccctaaata
     2161 atccttaaaa actccatttc cacccctccc agttcccaac tattttgtcc gcccacagcg
     2221 gggcattttt cttcctgtta tgtttttaat caaacatcct gccaactcca tgtgacaaac
     2281 cgtcatcttc ggctactttt tctctgtcac agaatgaaaa tttttctgtc atctcttcgt
     2341 tattaatgtt tgtaattgac tgaatatcaa cgcttatttg cagcctgaat ggcgaatggg
     2401 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2461 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2521 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2581 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     2641 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc ttaatagtgg
     2701 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     2761 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2821 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     2881 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2941 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     3001 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3061 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     3121 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     3181 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     3241 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3301 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3361 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     3421 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     3481 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     3541 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     3601 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     3661 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     3721 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     3781 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     3841 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     3901 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     3961 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4021 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4081 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4141 agagcgcaga taccaaatac tattcttcta gtgtagccgt agttaggcca ccacttcaag
     4201 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4261 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4321 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4381 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4441 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4501 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4561 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4621 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4681 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4741 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     4801 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tagaccagcc gcgtaacctg
     4861 gcaaaatcgg ttacggttga gtaataaatg gatgccctgc gtaagcgggt gtgggcggac
     4921 aataaagtct taaactgaac aaaatagatc taaactatga caataaagtc ttaaactaga
     4981 cagaatagtt gtaaactgaa atcagtccag ttatgctgtg aaaaagcata ctggactttt
     5041 gttatggcta aagcaaactc ttcattttct gaagtgcaaa ttgcccgtcg tattaaagag
     5101 gggcgtggcc aagggcatgg taaagactat attcgcggcg ttgtgacaat ttaccgaaca
     5161 actccgcggc cgggaagccg atctcggctt gaacgaattg ttaggtggcg gtacttgggt
     5221 cgatatcaaa gtgcatcact tcttcccgta tgcccaactt tgtatagaga gccactgcgg
     5281 gatcgtcacc gtaatctgct tgcacgtaga tcacataagc accaagcgcg ttggcctcat
     5341 gcttgaggag attgatgagc gcggtggcaa tgccctgcct ccggtgctcg ccggagactg
     5401 cgagatcata gatatagatc tcactacgcg gctgctcaaa cttgggca
//
