LOCUS       sequence-178987-        5433 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express GFP-tagged C-terminal region of Dsup in mammalian cells 
            transiently.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5433)
  AUTHORS   Hashimoto T, Horikawa DD, Saito Y, Kuwahara H, Kozuka-Hata H, Shin-I
            T, Minakuchi Y, Ohishi K, Motoyama A, Aizu T, Enomoto A, Kondo K, 
            Tanaka S, Hara Y, Koshikawa S, Sagara H, Miura T, Yokobori S, 
            Miyagawa K, Suzuki Y, Kubo T, Oyama M, Kohara Y, Fujiyama A, Arakawa
            K, Katayama T, Toyoda A, Kunieda T
  TITLE     Extremotolerant tardigrade genome and improved radiotolerance of 
            human cultured cells by tardigrade-unique protein.
  JOURNAL   Nat Commun. 2016 Sep 20;7:12808. doi: 10.1038/ncomms12808.
  PUBMED    27649274
REFERENCE   2  (bases 1 to 5433)
  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: "Nat 
            Commun."; date: "2016-09-20"; volume: "7:12808. doi"; pages: " 
            10.1038/ncomms12808"
COMMENT     Sequence Label: pAcGFP1-N1-Dsup-C
FEATURES             Location/Qualifiers
     source          1..5433
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..142
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        159..350
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     307..327
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(352..456)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      483..938
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(570..589)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     780..801
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    945..1066
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(956..975)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1010..1029
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1188..1907)
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     promoter        complement(2719..2922)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2748..2768)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2923..3226
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3401..3989)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3481..3500)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4318..4365
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4390..4409
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4597..5391)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4597..5391)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4708..4727)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5318..5337
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5426..5433,1..350))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg
       61 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
      121 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
      181 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg
      241 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
      301 ttccacaccc taactgacac acattccaca gctggttctt tccgcctcag gactcttcct
      361 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
      421 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
      481 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
      541 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
      601 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
      661 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
      721 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
      781 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
      841 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
      901 taacgcgaat tttaacaaaa tattaacgct tacaatttac gccttaagat acattgatga
      961 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     1021 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     1081 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     1141 cctctacaaa tgtggtatgg ctgattatga tctagagtcg cggccgctca cttgtacagc
     1201 tcatccatgc cgtgggtgat ggcggcggcg gtcacgaagc cgaagtagat catgtgatcg
     1261 cgcttctcgt tggggtcctt ggacagggcg ctctgggtgg acaggtagtg gttatcgggc
     1321 agcagcacag ggccatcgcc gatgggggta ttctgctggt agtggtcggc cagctgcacg
     1381 ctgccatcct cgatgttgtg gcggatcttg aagttcacct tgatgccatt cttggccttg
     1441 tcggtcatga tgtacacatt gtgggcgttg tagttgtact ccatcttatt gcccaggatg
     1501 ttgccatcct ccttgaaatc ggtgccggtc agctcgatgc gattcaccag ggtatcgccc
     1561 tcgaacttca cctcggcgcg cgacttgtag ttgccgtcat cctcgaagaa gatggtgcgc
     1621 tcctggatgt agccctcagg catggcgctc ttgaagaagt cgtgctgctt catgtgatcg
     1681 gggtagcgtg agaagcactg cacgccgtag ctcagggtgg tcaccagggt gggccagggc
     1741 acaggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgta ggtggcatcg
     1801 ccctcgccct cgccgctcac gctgaacttg tggccattca catcgccatt cagctcgatc
     1861 aggatgggca cgatgccggt gaacagctcg gcgcccttgc tcaccatgac cggtggatcc
     1921 ttcctcttcc gtcctccagc ctttcctccc ttcttcttgg cacctgcgcc attctgggga
     1981 gctttagagg ctgccgattt ggcagcagtt gctttggtgg cagcagcttt gccagcacct
     2041 ccagtcgctt gcttcttagc gttggaaggt gtactggagg aagaagcagc acgcttcgct
     2101 ccgcctttcg ctgcagctgg agcggaaccc gcccctgtac ctgaagacgt caggcgggac
     2161 gaccgccgag gcggctcagc ggacgtatca cccgcatccg ctttcttacc agcgccccgc
     2221 cctctgcctt tcttagctcc accctcactg cctgatgcac caccttcgcc aacggtcgcc
     2281 gcggaatccc tctggtccga gcctcccttg gaggaagcat ctgccatctc gatatctcct
     2341 gacgactgtt ctcctccctt ggaggattca gcaccgctct tggctccttt ggaggcggca
     2401 gcgccctttg cagcaccctt agcaactcct ttagcagctg ctggaggccg tccacgacca
     2461 cggcctttgg ctggagcagc tccttcgttg ccagagctgg attctgcttg gccagaagtc
     2521 tctccttcag acttcttgtt gtcaccacct tgtccagagc cacttccagc tttggagtcg
     2581 tccttcttgt cgccgctgtg ctcgccctct ttcttgtcgt ccgtcttgtc tcccatggta
     2641 ccgtcgactg cagaattcga agcttgagct cgagatctga gtccggtagc gctagcggat
     2701 ctgacggttc actaaaccag ctctgcttat atagacctcc caccgtacac gcctaccgcc
     2761 catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg
     2821 ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa
     2881 ccgctatcca cgcccattga tgtactgcca aaaccgcatc accatggtaa tagcgatgac
     2941 taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg
     3001 ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac
     3061 ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa
     3121 agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc
     3181 gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata
     3241 tgggctatga actaatgacc ccgtaattga ttactattaa taactaatgc atggcggtaa
     3301 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     3361 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3421 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3481 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     3541 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     3601 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     3661 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3721 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     3781 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     3841 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     3901 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     3961 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4021 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4081 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4141 agtaacctga ggctatggca gggcctgccg ccccgacgtt ggctgcgagc cctgggcctt
     4201 cacccgaact tggggggtgg ggtggggaaa aggaagaaac gcgggcgtat tggccccaat
     4261 ggggtctcgg tggggtatcg acagagtgcc agccctggga ccgaaccccg cgtttatgaa
     4321 caaacgaccc aacaccgtgc gttttattct gtctttttat tgccgtcata gcgcgggttc
     4381 cttccggtat tgtctccttc cgtgtttcag ttagcctccc cctagggtgg gcgaagaact
     4441 ccagcatgag atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga
     4501 agcccaacct ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg
     4561 tcgcttggtc ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg
     4621 atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc
     4681 agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata
     4741 gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac
     4801 catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat
     4861 gctcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag
     4921 atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt
     4981 cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc
     5041 agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg
     5101 cacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc
     5161 gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt gcagttcatt
     5221 cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg
     5281 gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct
     5341 ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga
     5401 tcctcatcct gtctcttgat cgatctttgc aaa
//
