LOCUS       sequence-346866-        6686 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Chlamydomonas reinhardtii RSP3 recombinant protein in 
            insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6686)
  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 6686)
  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-GST-TEV-RSP3(160-516)
FEATURES             Location/Qualifiers
     source          1..6686
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(85..90,91..107,108..113))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(509..542)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        590..681
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     606..623
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             724..1377
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             1420..1440
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    2656..2790
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2693..2712)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2747..2766
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2900..3065
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3249..3703
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3336..3355)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3545..3566
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3729..3833
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3834..3902,3903..4694)
                     /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(4052..4071)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4865..5453
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5354..5373
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5607..5624
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  5758..5982
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6049..6582)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt
       61 gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt caactgggtt
      121 tgtgccttca tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc
      181 gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag
      241 gcattggcgg ccttgctgtt cttctacggc aaggtgctgt gcacggatct gcccttgctt
      301 caggagatcg gtagacctcg gccgtcgcgg cgcttgccgg tggtgctgac cccggatgaa
      361 gtggttcgca tcctcggttt tctggaaggc gagcatcgtt tgttcgccca ggactctagc
      421 tatagttcta gtggttggct acagctttgt ttgtactatc aacaggttga actgctgatc
      481 aacagatcct ctacgcggcc gcggtaccat aacttcgtat agcatacatt atacgaagtt
      541 atctggtttt actagtatcg attcgcgacc tactccggaa tattaataga tcatggagat
      601 aattaaaatg ataaccatct cgcaaataaa taagtatttt actgttttcg taacagtttt
      661 gtaataaaaa aacctataaa tattccggat tattcatacc gtcccaccat cgggcgcgga
      721 tccatgtccc ctatacttgg ttattggaag attaagggtc tggtccaacc gacccgtctc
      781 ctgctggaat acctggaaga aaaatacgaa gagcatctgt acgagaggga tgagggtgat
      841 aaatggcgta ataaaaagtt tgaactgggt ttggaatttc cgaatcttcc ttattacatc
      901 gacggtgatg tgaagttgac ccaatcgatg gctattatca gatatattgc tgacaagcac
      961 aatatgcttg gtggatgccc gaaggagcgt gcagagattt ccatgttgga gggagccgtc
     1021 ctggacataa gatacggtgt ttcccgtatt gcatactcta aggactttga gactctcaaa
     1081 gtagatttcc tttcgaagct gccggaaatg ctcaagatgt tcgaggatag actctgtcat
     1141 aaaacatatc tgaacggaga tcacgtaacc cacccagact ttatgctgta tgacgcactt
     1201 gacgttgttc tgtatatgga ccccatgtgc ctcgatgcct ttccgaagct tgtgtgcttc
     1261 aaaaaacgta ttgaagcgat acctcaaata gataagtatc ttaaatcctc caaatacatc
     1321 gcctggcctt tgcaaggctg gcaagcaact ttcggaggag gcgaccaccc acctaaaagc
     1381 gacggcggtg gtagtggagg tggttctggt ggctcagcag aaaaccttta ttttcagggt
     1441 gcagctgccg aaccaattct ggaagttctc gtaggaaaag tgctcgaaca aggccttatg
     1501 gaggtgctcg aggaggagga attggcggcc atgagggcac atcaggagca ttttgaacag
     1561 attaggaatg cggaacttgt ggcaacacag aggatggagg cggctgagag aagaaaactg
     1621 gaggaaaagg agcgccgtat gcagcaagag agggagcgtg tagaacgcga gcgcgttgtc
     1681 cgccagaaag ttgcagcatc agcattcgct aggggatacc tctcaggaat agttaatacg
     1741 gttttcgata gactggtttc tagcggatat atatacgacc ccgtaatgag ggaggtggaa
     1801 acggctttca tgccgtggct gaaggagcaa gcaataggtt acctcgccag aggtgttgtt
     1861 gcgcgcaggg ttgtagataa gctggttgag gacgccgctg ctgcgctggc agcgaatagg
     1921 agtaccctcg cggataaagc tgcttctact gcggcaacag ttgatgcatg ggcagagcgc
     1981 caggcgaaaa tggaggctga acttcagggt aaagaattgg aagcagttag gaggagacct
     2041 actttcgttc tgcgtgaact taaacctgct gtagcgtctg ccgatgcggt cgaagcagca
     2101 gcggctgaac tgacagccca ggcggaagag gcagctaacg ccaaatggga ggcagataaa
     2161 gccgaggcag ctgagaaagc tcgtgcggaa gctgaggccg cggccgagga gcaaaaggcg
     2221 ttgctggaag aattggcagc aaccgccgca gcagaagcag aggagcgcgg cgaagaacca
     2281 cctgcggaac ctcctagcct tcccgacggc gtagaacctg tagatgtgga agcagaggta
     2341 gctaaggcgg tcgaggctgt tcctaagccg ccggttaaag aagttacgga cattgatatt
     2401 ctttcttaca tgatggataa aggagccatc acaaaagacg cgattatcca ggcccttgcg
     2461 gtccatgcgc tgggagacaa agcctacacg aaccacccgg ctttcgcaga ggcggaaggc
     2521 gcctaaaagc ttgtcgagaa gtactagagg atcataatca gccataccac atttgtagag
     2581 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     2641 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     2701 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     2761 ctcatcaatg tatcttatca tgtctggatc tgatcactgc ttgagcctag aagatccggc
     2821 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactatc
     2881 ataaccccta ggagatccga accagataag tgaaatctag ttccaaacta ttttgtcatt
     2941 tttaattttc gtattagctt acgacgctac acccagttcc catctatttt gtcactcttc
     3001 cctaaataat ccttaaaaac tccatttcca cccctcccag ttcccaacta ttttgtccgc
     3061 ccacagcggg gcatttttct tcctgttatg tttttaatca aacatcctgc caactccatg
     3121 tgacaaaccg tcatcttcgg ctactttttc tctgtcacag aatgaaaatt tttctgtcat
     3181 ctcttcgtta ttaatgtttg taattgactg aatatcaacg cttatttgca gcctgaatgg
     3241 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3301 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3361 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     3421 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     3481 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     3541 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3601 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3661 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg
     3721 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3781 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3841 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3901 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3961 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4021 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4081 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4141 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4201 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4261 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4321 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4381 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4441 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4501 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4561 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4621 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4681 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4741 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4801 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4861 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4921 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4981 gcttcagcag agcgcagata ccaaatacta ttcttctagt gtagccgtag ttaggccacc
     5041 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5101 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5161 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5221 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5281 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5341 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5401 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5461 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5521 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     5581 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     5641 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata gaccagccgc
     5701 gtaacctggc aaaatcggtt acggttgagt aataaatgga tgccctgcgt aagcgggtgt
     5761 gggcggacaa taaagtctta aactgaacaa aatagatcta aactatgaca ataaagtctt
     5821 aaactagaca gaatagttgt aaactgaaat cagtccagtt atgctgtgaa aaagcatact
     5881 ggacttttgt tatggctaaa gcaaactctt cattttctga agtgcaaatt gcccgtcgta
     5941 ttaaagaggg gcgtggccaa gggcatggta aagactatat tcgcggcgtt gtgacaattt
     6001 accgaacaac tccgcggccg ggaagccgat ctcggcttga acgaattgtt aggtggcggt
     6061 acttgggtcg atatcaaagt gcatcacttc ttcccgtatg cccaactttg tatagagagc
     6121 cactgcggga tcgtcaccgt aatctgcttg cacgtagatc acataagcac caagcgcgtt
     6181 ggcctcatgc ttgaggagat tgatgagcgc ggtggcaatg ccctgcctcc ggtgctcgcc
     6241 ggagactgcg agatcataga tatagatctc actacgcggc tgctcaaact tgggcagaac
     6301 gtaagccgcg agagcgccaa caaccgcttc ttggtcgaag gcagcaagcg cgatgaatgt
     6361 cttactacgg agcaagttcc cgaggtaatc ggagtccggc tgatgttggg agtaggtggc
     6421 tacgtctccg aactcacgac cgaaaagatc aagagcagcc cgcatggatt tgacttggtc
     6481 agggccgagc ctacatgtgc gaatgatgcc catacttgag ccacctaact ttgttttagg
     6541 gcgactgccc tgctgcgtaa catcgttgct gctgcgtaac atcgttgctg ctccataaca
     6601 tcaaacatcg acccacggcg taacgcgctt gctgcttgga tgcccgaggc atagactgta
     6661 caaaaaaaca gtcataacaa gccatg
//
