LOCUS       sequence-346862-        6263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Chlamydomonas reinhardtii RSP6 recombinant protein in 
            insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6263)
  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 6263)
  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-RSP6
FEATURES             Location/Qualifiers
     source          1..6263
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(520..525,526..542,543..548))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(944..977)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1025..1116
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1041..1058
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             1236..1247
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2668..2802
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2705..2724)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2759..2778
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2912..3077
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3261..3715
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3348..3367)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3557..3578
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3741..3845
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3846..3914,3915..4706)
                     /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(4064..4083)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4877..5465
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5366..5385
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5619..5636
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  5770..5994
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(join(6061..6263,1..331))
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 gatcatagat atagatctca ctacgcggct gctcaaactt gggcagaacg taagccgcga
       61 gagcgccaac aaccgcttct tggtcgaagg cagcaagcgc gatgaatgtc ttactacgga
      121 gcaagttccc gaggtaatcg gagtccggct gatgttggga gtaggtggct acgtctccga
      181 actcacgacc gaaaagatca agagcagccc gcatggattt gacttggtca gggccgagcc
      241 tacatgtgcg aatgatgccc atacttgagc cacctaactt tgttttaggg cgactgccct
      301 gctgcgtaac atcgttgctg ctgcgtaaca tcgttgctgc tccataacat caaacatcga
      361 cccacggcgt aacgcgcttg ctgcttggat gcccgaggca tagactgtac aaaaaaacag
      421 tcataacaag ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt
      481 ctggaccagt tgcgtgagcg catacgctac ttgcattaca gtttacgaac cgaacaggct
      541 tatgtcaact gggtttgtgc cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg
      601 ggcagcagcg aagtcgaggc atttctgtcc tggctggcga acgagcgcaa ggtttcggtc
      661 tccacgcatc gtcaggcatt ggcggccttg ctgttcttct acggcaaggt gctgtgcacg
      721 gatctgccct tgcttcagga gatcggtaga cctcggccgt cgcggcgctt gccggtggtg
      781 ctgaccccgg atgaagtggt tcgcatcctc ggttttctgg aaggcgagca tcgtttgttc
      841 gcccaggact ctagctatag ttctagtggt tggctacagc tttgtttgta ctatcaacag
      901 gttgaactgc tgatcaacag atcctctacg cggccgcggt accataactt cgtatagcat
      961 acattatacg aagttatctg gttttactag tatcgattcg cgacctactc cggaatatta
     1021 atagatcatg gagataatta aaatgataac catctcgcaa ataaataagt attttactgt
     1081 tttcgtaaca gttttgtaat aaaaaaacct ataaatattc cggattattc ataccgtccc
     1141 accatcgggc gcggatccat ggcagcggat gtcggtcagg ctctcgcatt cctgcagcag
     1201 gttaagacca cacaaggagc aagcatctat gagggattga aggccgcttt ggcgaaagtg
     1261 ctggaagatc gcccagtaaa cgcggttgag gctcttgaaa ccagcgtctt gtcaacgccg
     1321 cccgctgcca acttgtcagt tccgttggtc cctgcagcca gcgctgcggc agcagcggct
     1381 gcagttgcaa aggcatccct gtttggagat ccagaaccgg tattggatcc cgaaagtggt
     1441 gagcctatcg accctgacgc ccccaacgag tttgaatgtg aagacgtaga gggtgatggc
     1501 gacctgctgg atggactggg agttggcctg ggcaggcaag aaatgtatgc cgcaatgctg
     1561 gcagttaagc gcctgggcga ggacgcgaaa agaggtgtga gcacggtgag gttcttcgga
     1621 aaatttttcg gaacgcaagc tgattattat gtctttgaga ctacgttgca aagtaatcct
     1681 gatatgccgg aggcccctga aggaacgata ccacttgaac cttacggaga aggcgtcaac
     1741 gcatacatct actttgtatc aaatacactg ggcggtccgc ttcaacagtt accatacgtt
     1801 accccagaac agataaaagc aagtcgtctg cttagacgct acttgactgg cagacttgat
     1861 gcgcccgtta gtgctttccc tgcttttccc ggaaacgagg caaactactt gcgtgctctc
     1921 atcgcccgca taagcgcagc gactgtttgt tgtcccaggg gattttttac ggctgatgat
     1981 gattcagctg aactctcggc gaatgacgag tgggtccccc tcaaaggcag ggaaatggca
     2041 ctcccggtca attggagcca ccgctatgct catctcaagg gccagggccg cacagtcact
     2101 cataagaggg accctcccga cgaagaggag gaacctgaaa agaacttctg gaccgctgag
     2161 gaaatggaag ccggcccccc tccgcttgct actctcgaca ctgacgctcc tcttccggcc
     2221 gccacgggcg ataaggtccc gcctcccgcc tggagccccg tctttgcctc agcatcggtt
     2281 accaccagaa atcaagttgc tggtgtgcgt tccaaccgtt ggccgggagc ggtatgtgcc
     2341 tgtgctggcc gccatttcac gagtatgtat gtcggctggg gtatcaaggc tggcggagaa
     2401 tggtcacctt gcccccctcc tccccccgtg ccccaatggg gcgctcctgc agccggtgta
     2461 gaaggcggac agcaattgct gcttgaatgt aatgatcttc cccctaagcc tgcccctcca
     2521 gaagaggagg acgaataaaa gcttgtcgag aagtactaga ggatcataat cagccatacc
     2581 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     2641 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     2701 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2761 ggtttgtcca aactcatcaa tgtatcttat catgtctgga tctgatcact gcttgagcct
     2821 agaagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     2881 gcaataacta tcataacccc taggagatcc gaaccagata agtgaaatct agttccaaac
     2941 tattttgtca tttttaattt tcgtattagc ttacgacgct acacccagtt cccatctatt
     3001 ttgtcactct tccctaaata atccttaaaa actccatttc cacccctccc agttcccaac
     3061 tattttgtcc gcccacagcg gggcattttt cttcctgtta tgtttttaat caaacatcct
     3121 gccaactcca tgtgacaaac cgtcatcttc ggctactttt tctctgtcac agaatgaaaa
     3181 tttttctgtc atctcttcgt tattaatgtt tgtaattgac tgaatatcaa cgcttatttg
     3241 cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3301 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3361 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3421 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3481 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3541 gtccacgttc ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3601 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3661 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     3721 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3781 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3841 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3901 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3961 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4021 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4081 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4141 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4201 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4261 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4321 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4381 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4441 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4501 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4561 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4621 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4681 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4741 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4801 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4861 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4921 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4981 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tattcttcta gtgtagccgt
     5041 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5101 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5161 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5221 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5281 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5341 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5401 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5461 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5521 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     5581 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     5641 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     5701 tagaccagcc gcgtaacctg gcaaaatcgg ttacggttga gtaataaatg gatgccctgc
     5761 gtaagcgggt gtgggcggac aataaagtct taaactgaac aaaatagatc taaactatga
     5821 caataaagtc ttaaactaga cagaatagtt gtaaactgaa atcagtccag ttatgctgtg
     5881 aaaaagcata ctggactttt gttatggcta aagcaaactc ttcattttct gaagtgcaaa
     5941 ttgcccgtcg tattaaagag gggcgtggcc aagggcatgg taaagactat attcgcggcg
     6001 ttgtgacaat ttaccgaaca actccgcggc cgggaagccg atctcggctt gaacgaattg
     6061 ttaggtggcg gtacttgggt cgatatcaaa gtgcatcact tcttcccgta tgcccaactt
     6121 tgtatagaga gccactgcgg gatcgtcacc gtaatctgct tgcacgtaga tcacataagc
     6181 accaagcgcg ttggcctcat gcttgaggag attgatgagc gcggtggcaa tgccctgcct
     6241 ccggtgctcg ccggagactg cga
//
