LOCUS       sequence-178995-        5406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express GFP-tagged Dsup lacking C-terminal region in mammalian 
            cells transiently.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5406)
  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 5406)
  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-deltaC
FEATURES             Location/Qualifiers
     source          1..5406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        90..281
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     238..258
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(283..387)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      414..869
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(501..520)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     711..732
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    876..997
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(887..906)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     941..960
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1119..1838)
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     promoter        complement(2623..2826)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2652..2672)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2827..3130
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3305..3893)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3385..3404)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4222..4269
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4294..4313
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4501..5295)
                     /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(4501..5295)
                     /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(4612..4631)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5222..5241
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5330..5406,1..281))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5344..5406,1..73)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag
       61 ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca
      121 tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact
      181 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
      241 ctaactgaca cacattccac agctggttct ttccgcctca ggactcttcc tttttcaata
      301 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
      361 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc
      421 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
      481 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
      541 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
      601 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
      661 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
      721 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
      781 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
      841 ttttaacaaa atattaacgc ttacaattta cgccttaaga tacattgatg agtttggaca
      901 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
      961 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     1021 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     1081 atgtggtatg gctgattatg atctagagtc gcggccgctc acttgtacag ctcatccatg
     1141 ccgtgggtga tggcggcggc ggtcacgaag ccgaagtaga tcatgtgatc gcgcttctcg
     1201 ttggggtcct tggacagggc gctctgggtg gacaggtagt ggttatcggg cagcagcaca
     1261 gggccatcgc cgatgggggt attctgctgg tagtggtcgg ccagctgcac gctgccatcc
     1321 tcgatgttgt ggcggatctt gaagttcacc ttgatgccat tcttggcctt gtcggtcatg
     1381 atgtacacat tgtgggcgtt gtagttgtac tccatcttat tgcccaggat gttgccatcc
     1441 tccttgaaat cggtgccggt cagctcgatg cgattcacca gggtatcgcc ctcgaacttc
     1501 acctcggcgc gcgacttgta gttgccgtca tcctcgaaga agatggtgcg ctcctggatg
     1561 tagccctcag gcatggcgct cttgaagaag tcgtgctgct tcatgtgatc ggggtagcgt
     1621 gagaagcact gcacgccgta gctcagggtg gtcaccaggg tgggccaggg cacaggcagc
     1681 ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc gccctcgccc
     1741 tcgccgctca cgctgaactt gtggccattc acatcgccat tcagctcgat caggatgggc
     1801 acgatgccgg tgaacagctc ggcgcccttg ctcaccatga ccggtggatc cttcctcttc
     1861 cgtcctccag cctttcctcc cttcttcttg gcacctgcgc cattctgggg agctttagac
     1921 accacactgt ggtaagcatc gactaccttc tccgccacag tcgatgcagc cgacgatgcc
     1981 acatccttca cagtcgtggc tgcgttcttc acagcatcga ctgcagtatg catgatgcta
     2041 ggagcatcct cgacggcttt tcctgcgaca tctttggcgc ctgcaacgac gtctttgaca
     2101 gctccgatga cagagtgact cttgctactg tcggaagatg atttgccgct gtctttagac
     2161 tggccctgac tttgtccaga agatttggcg tcgctttgag acttggtgtc accttgggac
     2221 ttactatcgc caccagacgg gacttctttg gctgggcttt tggatttctc cttttggtca
     2281 gagctgaaag ttttcgggtc ggtaggtgta gtactgaact tctgtccctg agtactagaa
     2341 gagcccgctg tagtagagga atctttgcct tgggatcctc ctgtcttgac aatggcagct
     2401 gaggtggcgg aggaaccggt acctttggta acggttacgt tcttggagtc ttgcccgctc
     2461 ccttgcgaag catctttctt cgtttcctca gatttacctg tggaagaggg ttctgtggat
     2521 gattggtgtg tggatgccat ggtaccgtcg actgcagaat tcgaagcttg agctcgagat
     2581 ctgagtccgg tagcgctagc ggatctgacg gttcactaaa ccagctctgc ttatatagac
     2641 ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt
     2701 tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag
     2761 acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg
     2821 catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca
     2881 taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg
     2941 gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata
     3001 ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat
     3061 tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag
     3121 ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta
     3181 ttaataacta atgcatggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3241 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3301 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3361 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3421 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3481 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3541 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3601 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3661 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3721 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3781 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3841 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3901 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3961 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4021 ttaaatcaat ctaaagtata tatgagtaac ctgaggctat ggcagggcct gccgccccga
     4081 cgttggctgc gagccctggg ccttcacccg aacttggggg gtggggtggg gaaaaggaag
     4141 aaacgcgggc gtattggccc caatggggtc tcggtggggt atcgacagag tgccagccct
     4201 gggaccgaac cccgcgttta tgaacaaacg acccaacacc gtgcgtttta ttctgtcttt
     4261 ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt tcagttagcc
     4321 tccccctagg gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc
     4381 cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga
     4441 aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc
     4501 tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat
     4561 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg
     4621 ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa
     4681 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac
     4741 gacgagatcc tcgccgtcgg gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc
     4801 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt
     4861 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag
     4921 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg
     4981 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc
     5041 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg
     5101 cgctgcctcg tcttgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac
     5161 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg
     5221 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc
     5281 atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcgatct ttgcaaaagc
     5341 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct
     5401 cggcct
//
