LOCUS       sequence-346865-        6344 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Chlamydomonas reinhardtii RSP4 recombinant protein in 
            insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6344)
  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 6344)
  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-HISX6-TEV-RSP4
FEATURES             Location/Qualifiers
     source          1..6344
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(536..541,542..558,559..564))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(960..993)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        1041..1132
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1057..1074
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             1187..1204
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1217..1237
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    2765..2899
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2802..2821)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2856..2875
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3009..3174
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3358..3812
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3445..3464)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3654..3675
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3838..3942
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3943..4011,4012..4803)
                     /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(4161..4180)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4974..5562
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5463..5482
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5716..5733
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  5867..6091
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(join(6158..6344,1..347))
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 tcgccggaga ctgcgagatc atagatatag atctcactac gcggctgctc aaacttgggc
       61 agaacgtaag ccgcgagagc gccaacaacc gcttcttggt cgaaggcagc aagcgcgatg
      121 aatgtcttac tacggagcaa gttcccgagg taatcggagt ccggctgatg ttgggagtag
      181 gtggctacgt ctccgaactc acgaccgaaa agatcaagag cagcccgcat ggatttgact
      241 tggtcagggc cgagcctaca tgtgcgaatg atgcccatac ttgagccacc taactttgtt
      301 ttagggcgac tgccctgctg cgtaacatcg ttgctgctgc gtaacatcgt tgctgctcca
      361 taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc gaggcataga
      421 ctgtacaaaa aaacagtcat aacaagccat gaaaaccgcc actgcgccgt taccaccgct
      481 gcgttcggtc aaggttctgg accagttgcg tgagcgcata cgctacttgc attacagttt
      541 acgaaccgaa caggcttatg tcaactgggt ttgtgccttc atccgtttcc acggtgtgcg
      601 tcacccggca accttgggca gcagcgaagt cgaggcattt ctgtcctggc tggcgaacga
      661 gcgcaaggtt tcggtctcca cgcatcgtca ggcattggcg gccttgctgt tcttctacgg
      721 caaggtgctg tgcacggatc tgcccttgct tcaggagatc ggtagacctc ggccgtcgcg
      781 gcgcttgccg gtggtgctga ccccggatga agtggttcgc atcctcggtt ttctggaagg
      841 cgagcatcgt ttgttcgccc aggactctag ctatagttct agtggttggc tacagctttg
      901 tttgtactat caacaggttg aactgctgat caacagatcc tctacgcggc cgcggtacca
      961 taacttcgta tagcatacat tatacgaagt tatctggttt tactagtatc gattcgcgac
     1021 ctactccgga atattaatag atcatggaga taattaaaat gataaccatc tcgcaaataa
     1081 ataagtattt tactgttttc gtaacagttt tgtaataaaa aaacctataa atattccgga
     1141 ttattcatac cgtcccacca tcgggcgcgg atccatgggc agcagccatc atcatcatca
     1201 tcacggtggc tcagcagaaa acctttattt tcagggtatg gctgcggtcg attccgtggc
     1261 gcaggccctt gcttatttgc aagtacatag tccacaagac ggaacgtcga tgtatgatca
     1321 cctggtaaag ctcgtgagta aagttctgga ggaccagccc aagaacgccg ttgaccttct
     1381 ggaaacctcc ctccttgtga agaaatccac gttcgatcca aaagaatcaa gcccacttgt
     1441 gccaatacca gtcgcccccg acgctactca aactcaagcg gccgtttcga tatttggcga
     1501 cccagagttg cctataaatc cggcaacagg agagcctgtt ccggcagatc ccccaaacga
     1561 gtttgaggct gagaatatgc tgggtgctgc ggcggttctg gattgtctcg gcgttggcct
     1621 tggaagagag ttgggcgtca acatcgcgtt ggccgctaaa cgtattggcg aggacccaaa
     1681 gctcgcagtt agatcggttc gttttttcgg taaatttctg ggtttgtata gtgattattt
     1741 cgtgtttgag gttgcattta aaaaagaggc tgccaaggag gctgcccctg ctgcaccagc
     1801 gcccgaaagg gttgagggcg aagccgcctc atcatctgct cccgaggtcc ccgtagagga
     1861 gccgggtaag ggtgcgaaca aatttacgta tttggtgtgt tccagtttgg gtggccccct
     1921 gacgcgcctc ccagatgtaa ccccagctca agttaaggcc tcacgtcgca tcaaaaaact
     1981 tttgaccggc aggcttacat cacacgtttc aacttatcca gctttcccag gaaatgaggc
     2041 gaactacctt cgtgcgctca ttgccaggat ttctgccgca acagtcgttg caccaagtga
     2101 cctgttttcc ctcaatgatg aaactggtga gttggaacgc gcggaagatt gggagccgcc
     2161 cgcgggccgc gagatggcgg ctcctacggc atgggtgcat gtcagacccc acctcaaatc
     2221 ccaaggccgt tgcgaagtac acaaaaggga actccccgaa gacgctgatg aagatgagtt
     2281 ttataatgag gatgaactcg aagaaggtcc tgaccttctt gcggcattgg aggaagacgc
     2341 ccaactccct ggtgagcaag ccgcatggac gccgatctac agtagcgcct cggaggcggt
     2401 caagacccaa gcgggcggcc tcagatcact cgtctggccg ggagcggttt gcggtggtcg
     2461 tggatccgaa tggacctgcg tttacgttgg ctggggagta aaaaacgcgc cgttcgttcc
     2521 gctcccgccg ccgcccgtgg cccaggaatt tgcgtggggt gaagtcgaga ctcaggaact
     2581 cgaactgaaa cccgcgccac cgcctccgga ggaagaggca gaagcagacg agtaaaagct
     2641 tgtcgagaag tactagagga tcataatcag ccataccaca tttgtagagg ttttacttgc
     2701 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     2761 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     2821 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     2881 atcttatcat gtctggatct gatcactgct tgagcctaga agatccggct gctaacaaag
     2941 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactatca taacccctag
     3001 gagatccgaa ccagataagt gaaatctagt tccaaactat tttgtcattt ttaattttcg
     3061 tattagctta cgacgctaca cccagttccc atctattttg tcactcttcc ctaaataatc
     3121 cttaaaaact ccatttccac ccctcccagt tcccaactat tttgtccgcc cacagcgggg
     3181 catttttctt cctgttatgt ttttaatcaa acatcctgcc aactccatgt gacaaaccgt
     3241 catcttcggc tactttttct ctgtcacaga atgaaaattt ttctgtcatc tcttcgttat
     3301 taatgtttgt aattgactga atatcaacgc ttatttgcag cctgaatggc gaatgggacg
     3361 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3421 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3481 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3541 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3601 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttctta atagtggact
     3661 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3721 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3781 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     3841 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3901 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3961 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4021 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4081 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4141 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     4201 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     4261 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     4321 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     4381 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     4441 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     4501 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4561 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     4621 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     4681 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     4741 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     4801 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     4861 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4921 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4981 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5041 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5101 gcgcagatac caaatactat tcttctagtg tagccgtagt taggccacca cttcaagaac
     5161 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5221 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5281 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5341 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5401 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5461 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5521 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5581 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5641 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     5701 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     5761 tttctcctta cgcatctgtg cggtatttca caccgcatag accagccgcg taacctggca
     5821 aaatcggtta cggttgagta ataaatggat gccctgcgta agcgggtgtg ggcggacaat
     5881 aaagtcttaa actgaacaaa atagatctaa actatgacaa taaagtctta aactagacag
     5941 aatagttgta aactgaaatc agtccagtta tgctgtgaaa aagcatactg gacttttgtt
     6001 atggctaaag caaactcttc attttctgaa gtgcaaattg cccgtcgtat taaagagggg
     6061 cgtggccaag ggcatggtaa agactatatt cgcggcgttg tgacaattta ccgaacaact
     6121 ccgcggccgg gaagccgatc tcggcttgaa cgaattgtta ggtggcggta cttgggtcga
     6181 tatcaaagtg catcacttct tcccgtatgc ccaactttgt atagagagcc actgcgggat
     6241 cgtcaccgta atctgcttgc acgtagatca cataagcacc aagcgcgttg gcctcatgct
     6301 tgaggagatt gatgagcgcg gtggcaatgc cctgcctccg gtgc
//
