LOCUS       sequence-378564-        7008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of RvCAHS3 (fly codon 
            optimized)-GS_linker-AcGFP1 in Drosophila cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7008)
  AUTHORS   Tanaka A, Nakano T, Watanabe K, Masuda K, Honda G, Kamata S, Yasui 
            R, Kozuka-Hata H, Watanabe C, Chinen T, Kitagawa D, Sawai S, Oyama 
            M, Yanagisawa M, Kunieda T
  TITLE     Stress-dependent cell stiffening by tardigrade tolerance proteins 
            that reversibly form a filamentous network and gel.
  JOURNAL   PLoS Biol. 2022 Sep 6;20(9):e3001780. doi: 
            10.1371/journal.pbio.3001780. eCollection 2022 Sep.
  PUBMED    36067153
REFERENCE   2  (bases 1 to 7008)
  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"; journalName: "PLoS 
            Biol."; date: "2022-09-6"; volume: "20(9):e3001780. doi"; pages: " 
            10.1371/journal.pbio.3001780. eCollection 2022 Sep"
COMMENT     Sequence Label: pAc5.1/V5-HisA-RvCAHS3-GS-AcGFP1
FEATURES             Location/Qualifiers
     source          1..7008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2565
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             3565..4284
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     CDS             4314..4355
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4365..4382
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4405..4422)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4455..4589
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4479..4498)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4533..4552
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(5190..5778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5270..5289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5949..6740,6741..6809))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6572..6591
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6810..6914)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6982..7000
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cttcaagaat taattctata ttctaaaaac acaaatgata cttctaaaaa aaaatcatga
       61 atggcatcaa ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac
      121 atcatttttc cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg
      181 ccattaaagc tggctgatcg tcgccaaaca ccaaatacat aatgaatatg tacacattcg
      241 agaaagaagc gatcaaagaa gcgtcttcgg gcggagtagg agaatgcgga ggagaaggag
      301 aacgagctga tctagtatct ctccacaatc caatgccaac tgaccaactg gccatattcg
      361 gagcaatttg aagccaattt ccatcgcctg gcgatcgctc cattcttggc tatatgtttt
      421 tcaccgttac ccggggccat tttcaaagac tcgtcggcaa gataagattg tgtcactcgc
      481 tgtctctctt catttgtcga agaatgctga ggaatttcgc gatgacgtcg gcgagtattt
      541 tgaagaatga gaataatttg tatttatacg aaaatcagtt agtggaattt tctacaaaaa
      601 catgttatct atagataatt ttgttgcaaa atatgttgac tatgacaaag attgtatgta
      661 tataccttta atgtattctc attttcttat gtatttataa tggcaatgat gatactgatg
      721 atattttaag atgatgccag accaaaaggc ttgaatttct gcgtcttttg ccgaacgcag
      781 tgcatgtgca attgttgttt tttggaatat tcaattttcg gactgtccgc tttgatttca
      841 gtttcttggc ttattcaaaa agcaaagtaa agccaaaaaa gcgagatggc aataccaaat
      901 gcggcaaaac ggtagtggaa ggaaaggggt gcggggcagc ggaaggaagg gtggggcggg
      961 gcgtggcggg gtctgtggct gggcgcgacg tcaccgacgt tggagccact cctttgacca
     1021 tgtgtgcgtg tgtgtattat tcgtgtctcg ccactcgccg gttgtttttt tctttttatg
     1081 ctgcgctctc tctagcgcca tctcgcttac gcatgctcaa cgcaccgcat gttgccgttt
     1141 ccttttatgc gtcattttgg ctcgaaatag gcaattattt aaacaaagat tagtcaacga
     1201 aaacgctaaa ataaataagt ctacaatatg gttacttatt gccatgtgtg tgcagccaac
     1261 gatagcaaca aaagcaacaa cacaggtggc tttccctctt tcactttttg tttgcaagcc
     1321 gcgtgcgagc aagacggcac gaccggcaaa cgcaattacg ctgacaaaga gcagacgaag
     1381 ttttggcgaa aaacatcaag gcgcctgata cgaatgcatt tgcaataaca attgcgatat
     1441 ttaatattgt ttatgaagct gtttgacttc aaaacacaca aaaaaaaaaa taaaacaaat
     1501 tatttgaaag agaattagga atcggacgct tatcgttagg gtaacaacaa gaaatgctta
     1561 ctgagtcaca gcctctggaa aactgccgca agccagagag agagagaaaa agagggagag
     1621 cagcttagac cgcatgtgct tgtgtgtgag gcgtctctct cttcgtctct gttgcgcaaa
     1681 cgcatagact gcactgaaaa aatcgattac ctatttttta tgaatgaata tttgcactat
     1741 tactattcaa aactattaag atagcaatca cattcaatag ccaaatacta taccacctga
     1801 gcgatgcaac gaaatgatca atttgagcaa aaatgctgca tatttaggac ggcatcatta
     1861 tagaaatgct tcttgctgtg tacttttctc tcgtctggca gctgtttcgc cgttattgtt
     1921 aaaaccggct taagttaggt gtgttttcta cgactagtga atgccctact agaagatgtg
     1981 tgttgcacaa aatgtccctg gaataaccaa tttgaagtgc agatagcagt aaacgtaagc
     2041 taatatgaat attatttaac tgtaatgttt taatatcgct ggacattact aataaaccca
     2101 ctataaacac atgtacatat gtatgttttg gcatacaatg agtagttggg gaaaaaatgt
     2161 gtaaaagcac cgtgaccatc acagcataaa gataaccagc tgaagtatcg aatatgagta
     2221 acccccaaat tgaatcacat gccgcaactg ataggaccca tggaagtaca ctcttcatgg
     2281 cgatatacaa gacacacaca agcacgaaca cccagttgcg gaggaaattc tccgtaaatg
     2341 aaaacccaat cggcgaacaa ttcataccca tatatggtaa aagttttgaa cgcgacttga
     2401 gagcggagag cattgcggct gataaggttt tagcgctaag cgggctttat aaaacgggct
     2461 gcgggaccag ttttcatatc actaccgttt gagttcttgt gctgtgtgga tactcctccc
     2521 gacacaaagc cgctccatca gccagcagtc gtctaatcca gagaccccgg atcggggtac
     2581 ctactagtcc agtgtggtgg aattgccacc atgagtagcc gccagaatca gcagagcagc
     2641 agccagcata gtagcagctc ccagcaaggc ggacaaggtg gacagggtgt tcaaggcagc
     2701 agcagctata gccgcaccga ggttcacaca agcagcggcg gaccaacaat tggaggtgcc
     2761 caacgtacag tgccagtgcc accaggatcg catagcgagg tgcacgaaga acgcgaagtg
     2821 atcaagcacg gcaccaagac cgagagcgag acacacgttg tgaccgtgcc agtgacaacc
     2881 ttcggcagca ccaacatgga aagcgtgcgc accggattca ccgtgacgca ggataagaac
     2941 ctgaccgtgg ccgctccgaa tatcgccgct ccaatccaca gcaacctgga tctgaatctc
     3001 ggaggcggag cccgtgccga aattaccgct ggaacaacgg tggatctgag caagatccag
     3061 cgcaaggatc tgggccccga agagtacgcc cgttacaagg ccaaggtgga acagctggcc
     3121 cgccaggatg aacaggatgc tggaatgcgt gccgcgcagt accgtgaaga ggtggaacgt
     3181 gatgccgagc tgatccgcca gattctggaa cgccagcaca tccgcgattt ggagttccgc
     3241 aaagagatgg tcgagaatca agtgaaccgc caagagcgcg agatccagtt ggaggccgaa
     3301 tatgccatgc gcgccctgga actggaacgc aatgccgcca aagaagccct ggaaagcgcc
     3361 aaggctcaga ccaacgtgaa cgtgaaggtc gagagcgcca tcggaaccac agtgtccaag
     3421 ggcgccattc agaccagcgc cgataagagc agcaccacca agacaggacc caccaccgtg
     3481 acacagatta agcacaccga gcagcatacc gagcgccgcg gtggaggcgg ttcaggcgga
     3541 ggtggctctg gcggtggcgg atcgatggtg agcaagggcg ccgagctgtt caccggcatc
     3601 gtgcccatcc tgatcgagct gaatggcgat gtgaatggcc acaagttcag cgtgagcggc
     3661 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     3721 aagctgcctg tgccctggcc caccctggtg accaccctga gctacggcgt gcagtgcttc
     3781 tcacgctacc ccgatcacat gaagcagcac gacttcttca agagcgccat gcctgagggc
     3841 tacatccagg agcgcaccat cttcttcgag gatgacggca actacaagtc gcgcgccgag
     3901 gtgaagttcg agggcgatac cctggtgaat cgcatcgagc tgaccggcac cgatttcaag
     3961 gaggatggca acatcctggg caataagatg gagtacaact acaacgccca caatgtgtac
     4021 atcatgaccg acaaggccaa gaatggcatc aaggtgaact tcaagatccg ccacaacatc
     4081 gaggatggca gcgtgcagct ggccgaccac taccagcaga atacccccat cggcgatggc
     4141 cctgtgctgc tgcccgataa ccactacctg tccacccaga gcgccctgtc caaggacccc
     4201 aacgagaagc gcgatcacat gatctacttc ggcttcgtga ccgccgccgc catcacccac
     4261 ggcatggatg agctgtacaa gtgaggccgc tcgagtctag agggcccttc gaaggtaagc
     4321 ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat caccatcacc
     4381 attgagttta aacccgctga tcagcctcga ctgtgccttc taaggcctga gctcgctgat
     4441 cagcctcgat cgaggatcca gacatgataa gatacattga tgagtttgga caaaccacaa
     4501 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     4561 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     4621 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     4681 tggctgatta tgatcagtcg accgatgccc ttgagagcct tcaacccagt cagctccttc
     4741 cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa
     4801 ctcgtaggac aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg
     4861 agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa
     4921 gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc
     4981 atggcggccg acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc
     5041 ttccccatta tgattcttct cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg
     5101 ctgtccaggc aggtagatga cgaccatcag ggacagcttc aaggccagca aaaggccagg
     5161 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5221 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5281 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5341 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5401 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5461 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5521 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5581 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     5641 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5701 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5761 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5821 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5881 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5941 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6001 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6061 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6121 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6181 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6241 ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg
     6301 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6361 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6421 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6481 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6541 ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta
     6601 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6661 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6721 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6781 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6841 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6901 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     6961 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgt
//
