LOCUS       sequence-346864-        7139 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Rescue of RSP6 Chlamydomonas mutant with tagged RSP6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7139)
  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 7139)
  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: pBR9-genomic_RSP6-HAX3
FEATURES             Location/Qualifiers
     source          1..7139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1641..1721
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     primer_bind     complement(1978..1997)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1979..1997)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2007..2024)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2007..2023)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2016..2038)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2165..2620
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2252..2271)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2462..2483
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2646..2750
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2751..2819,2820..3611)
                     /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(2969..2988)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3782..4370
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4271..4290
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4524..4541
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4658..4679
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4694..4699,4700..4717,4718..4724)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4732..4748
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     4737..4759
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4756..4772
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4756..4772
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4791..4811
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4793..4811
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             6056..6121
                     /codon_start=1
                     /product="2A peptide from foot-and-mouth disease virus 
                     polyprotein"
                     /label=F2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="VKQTLNFDLLKLAGDVESNPGP"
ORIGIN
        1 gggcggggtg gagcgggggt gcggcggggc ggtctgtacg gcttgagcga ttggacgcga
       61 gatggcacgg ggtgcaatgg accccggcca ttggggcgcg cgtacgcaga ctgtggggcg
      121 atgagagagt acgtgggcgt gcggagaagg cgggggctgt gacgattgac gaggggtgcg
      181 gagaggggca cggcaggaag tccgaatagg aatctgggaa gcgctgaggg cgctggaagc
      241 gctttacgtc aacgggactg aaggatgcct tggcctggcc tcccggctgc cgccgtgcag
      301 gggcgtatct acggtgcgct tcttcggcaa gttcttcggc acgcaggccg actactacgt
      361 gtttgagacc acgctgcaga gcaacccgga catgcccgag gcaccaggtg agcgtcgctg
      421 cggagggcgg cggtctgcgt gtggaggagg cggcttaggc ctacagcggt gctagaggga
      481 ccgcctaccg ggcaaagggg ttgcgccttc gcgcacgcct caatatcgta cagctgtcct
      541 ccataaccat gccccctagc tcctctccag cattgggcct gcgttatgac ggtcgccttg
      601 ccccacctcc cccatcctac taaagagggc accatcccct tggagccgta cggcgagggc
      661 gtcaacgcct acatctactt tgtgtccaac acgctgggcg gcccgctgca gcagctgccc
      721 tacgtcacgc ccgagcaggt gcgtgggcga gcgccaccgc acaccgcgac gcgattgtgt
      781 gtgtagtgtg tatgcttgtg gctgcgtggc gtgtttgggc cctagccgcc ctgtgtgccg
      841 cagacccgtt aacatgaccc cgcaccgcaa actcacgccg ttccacccac ctccacctcc
      901 caccctccaa gatcaaggcc agccgcctgc tgcgccgcta cctgaccggc cgcctggacg
      961 cgcccgtgtc cgccttccca gccttccccg gcaacgaggc caactacctg cgcgccctca
     1021 ttgcgcgcat cagcgccgcc accgtgtgct gcccgcgcgg cttcttcacc gcggacgacg
     1081 acagcgccga gctgtccgcc aacgacgagt gggtgccgct caagggtcgc gagatggcgc
     1141 tgcccgtcaa ctggtcgcac cgctacgcgc acctcaaggg ccagggccgc accgtgacgc
     1201 acaagcgcga cccgcccgac gaggaggagg agccggagaa gaacttctgg acggccgagg
     1261 agatggaggc cggcccgccg ccgctggcca ccctggacac ggacgcgccg ctgccggccg
     1321 ccactggcga caaggtgcca ccgcccgcct ggagccccgt gttcgccagc gcctcggtga
     1381 ccacgcgcaa ccaggtggcg ggcgtgcgct ccaaccgctg gccgggcgcg gtgtgcgcct
     1441 gcgcgggccg ccacttcacg tccatgtacg tgggctgggg catcaaggcc ggcggcgagt
     1501 ggtcgccctg cccgccgccg ccgcccgtgc cgcagtgggg cgcccccgct gctggcgtcg
     1561 agggcgggca gcagctgctg ctcgagtgca acgacctgcc gcccaagccc gcgccgccgg
     1621 aggaggagga cgaggaattc tacccttacg acgtcccgga ctacgcttac ccctacgatg
     1681 tgcctgacta cgcctacccc tacgatgtcc cggattacgc ttaaggatcc ccgctccgtg
     1741 taaatggagg cgctcgttga tctgagcctt gccccctgac gaacggcggt ggatggaaga
     1801 tactgctctc aagtgctgaa gcggtagctt agctccccgt ttcgtgctga tcagtctttt
     1861 tcaacacgta aaaagcggag gagttttgca attttgttgg ttgtaacgat cctccgttga
     1921 ttttggcctc tttctccatg ggcgggctgg gcgtatttga agcgggtacc caattcgccc
     1981 tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa
     2041 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     2101 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     2161 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2221 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2281 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2341 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2401 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2461 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2521 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2581 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     2641 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2701 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2761 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2821 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2881 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2941 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     3001 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3061 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3121 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3181 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3241 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     3301 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3361 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3421 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3481 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3541 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3601 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3661 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3721 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3781 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3841 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3901 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     3961 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4021 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4081 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4141 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4201 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4261 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4321 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4381 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4441 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4501 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     4561 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     4621 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     4681 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     4741 gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc gcaattaacc
     4801 ctcactaaag ggaacaaaag ctggagctcc accgcggtgg cggccgctct agacggcggg
     4861 gagctcgctg aggcttgaca tgattggtgc gtatgtttgt atgaagctac aggactgatt
     4921 tggcgggcta tgagggcggg ggaagctctg gaagggccgc gatggggcgc gcggcgtcca
     4981 gaaggcgcca tacggcccgc tggcggcacc catccggtat aaaagcccgc gaccccgaac
     5041 ggtgacctcc actttcagcg acaaacgagc acttatacat acgcgactat tctgccgcta
     5101 tacataacca ctcagctagc ttaagatccc atcaagcttg catgccgggc gcgccagaag
     5161 gagcgcagcc aaaccaggat gatgtttgat ggggtatttg agcacttgca acccttatcc
     5221 ggaagccccc tggcccacaa aggctaggcg ccaatgcaag cagttcgcat gcagcccctg
     5281 gagcggtgcc ctcctgataa accggccagg gggcctatgt tctttacttt tttacaagag
     5341 aagtcactca acatcttaaa atggccaggt gagtcgacga gcaagcccgg cggatcaggc
     5401 agcgtgcttg cagatttgac ttgcaacgcc cgcattgtgt cgacgaaggc ttttggctcc
     5461 tctgtcgctg tctcaagcag catctaaccc tgcgtcgccg tttccatttg caggatggcc
     5521 atgcatatgg ccaagctgac cagcgccgtt ccggtgctca ccgcgcgcga cgtcgccgga
     5581 gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga ggacgacttc
     5641 gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga ccaggtgagt
     5701 cgacgagcaa gcccggcgga tcaggcagcg tgcttgcaga tttgacttgc aacgcccgca
     5761 ttgtgtcgac gaaggctttt ggctcctctg tcgctgtctc aagcagcatc taaccctgcg
     5821 tcgccgtttc catttgcagg accaggtggt gccggacaac accctggcct gggtgtgggt
     5881 gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga
     5941 cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct
     6001 gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggacg ccccggtgaa
     6061 gcagaccctg aacttcgacc tgctgaagct ggcgggcgac gtggagagca acccgggccc
     6121 cctcgagatg gccgcggatg tgggccaggc tctggccttc ctgcagcaag tgaagacaac
     6181 acaaggcgca tcaatctatg aaggccttaa agcagcgttg gcgaaggttc tggaggatcg
     6241 gcctgtgaac gcggtcgagg cgctggagac ctcggtgctg tcgacacccc ccgcagcgaa
     6301 cctgagtgtg ccccttgttc ccgcagctgt gagtgacaac cgattcgttt ggccgcacgg
     6361 gtggtgtacg tagtagtgtc atgtgtggtg ccgtctcacg tacctgcagc ctgcctgaca
     6421 cgctggtgtg tgttaccgtg cttcgtcgct gtgcagtcag ctgcagctgc ggcagctgcc
     6481 gtggcgaagg cgagcctatt cgggtaggcc ctaagcttcg ggactgggtc ctctgaggag
     6541 tagctatgag attaggcaac tggcagggaa tgtgattgcc gaatggggga cgcgtgcacc
     6601 caaggtgaaa aggaggttgg cctatctggc cccgcggccg tggcccgcgc tcgttcccct
     6661 ggtgcacgcc ccggcttgac gcccttatcg cgtgctcgct gcagggaccc agagccagtg
     6721 cttgaccccg agtcagggga acctatcgat ccggatgctc ccaacgaggt ggggggcttg
     6781 cgagactggg gtgcaacagg tggtggcaac tacagtattg catggcgtta ggggtgcgca
     6841 acacgggggc cgccgcacag atgatagaaa ccaggcgttc ttgtaaactg cacgacccgc
     6901 gtgcctgcat tgcgccatga tccgggccgg gcattggttt gagggtgacc tcgccctctg
     6961 tcccttcccc cgatcgtgct taatccagtt cgagtgcgag gacgtggagg gcgacggtga
     7021 cctgctggac ggcctgggtg tggggctggg ccggcaggag atgtacgcgg ccatgctggc
     7081 ggtcaagagg ctgggggagg acgccaagag gtgcgaggcg gccgggcggg cgcgggccg
//
