LOCUS       sequence-178858-        6051 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express GFP-tagged Dsup in mammalian cells transiently.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6051)
  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 6051)
  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
FEATURES             Location/Qualifiers
     source          1..6051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        74..431
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        74..265
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(97..117)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      282..417
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             466..1260
                     /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             466..1260
                     /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(520..539)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1130..1149
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1448..1467)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1492..1539
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1868..2456
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2357..2376
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2631..2934
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2935..3138
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3089..3109
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4568..5287
                     /codon_start=1
                     /product="Aequorea coerulescens GFP"
                     /label=AcGFP1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
     polyA_signal    5409..5530
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5446..5465)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5500..5519
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5537..5992)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5674..5695)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5886..5905
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(6019..6051,1..72)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
       61 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
      121 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      181 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
      241 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
      301 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
      361 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
      421 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
      481 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
      541 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
      601 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
      661 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
      721 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
      781 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
      841 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
      901 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
      961 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     1021 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     1081 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     1141 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     1201 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     1261 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     1321 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     1381 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     1441 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     1501 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     1561 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     1621 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     1681 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     1741 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     1801 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     1861 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     1921 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     1981 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     2041 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     2101 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     2161 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     2221 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     2281 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     2341 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     2401 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     2461 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     2521 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
     2581 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     2641 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     2701 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     2761 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     2821 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     2881 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     2941 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     3001 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     3061 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     3121 gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggactcag
     3181 atctcgagct caagcttcga attctgcagt cgacggtacc atggcatcca cacaccaatc
     3241 atccacagaa ccctcttcca caggtaaatc tgaggaaacg aagaaagatg cttcgcaagg
     3301 gagcgggcaa gactccaaga acgtaaccgt taccaaaggt accggttcct ccgccacctc
     3361 agctgccatt gtcaagacag gaggatccca aggcaaagat tcctctacta cagcgggctc
     3421 ttctagtact cagggacaga agttcagtac tacacctacc gacccgaaaa ctttcagctc
     3481 tgaccaaaag gagaaatcca aaagcccagc caaagaagtc ccgtctggtg gcgatagtaa
     3541 gtcccaaggt gacaccaagt ctcaaagcga cgccaaatct tctggacaaa gtcagggcca
     3601 gtctaaagac agcggcaaat catcttccga cagtagcaag agtcactctg tcatcggagc
     3661 tgtcaaagac gtcgttgcag gcgccaaaga tgtcgcagga aaagccgtcg aggatgctcc
     3721 tagcatcatg catactgcag tcgatgctgt gaagaacgca gccacgactg tgaaggatgt
     3781 ggcatcgtcg gctgcatcga ctgtggcgga gaaggtagtc gatgcttacc acagtgtggt
     3841 gggagacaag acggacgaca agaaagaggg cgagcacagc ggcgacaaga aggacgactc
     3901 caaagctgga agtggctctg gacaaggtgg tgacaacaag aagtctgaag gagagacttc
     3961 tggccaagca gaatccagct ctggcaacga aggagctgct ccagccaaag gccgtggtcg
     4021 tggacggcct ccagcagctg ctaaaggagt tgctaagggt gctgcaaagg gcgctgccgc
     4081 ctccaaagga gccaagagcg gtgctgaatc ctccaaggga ggagaacagt cgtcaggaga
     4141 tatcgagatg gcagatgctt cctccaaggg aggctcggac cagagggatt ccgcggcgac
     4201 cgttggcgaa ggtggtgcat caggcagtga gggtggagct aagaaaggca gagggcgggg
     4261 cgctggtaag aaagcggatg cgggtgatac gtccgctgag ccgcctcggc ggtcgtcccg
     4321 cctgacgtct tcaggtacag gggcgggttc cgctccagct gcagcgaaag gcggagcgaa
     4381 gcgtgctgct tcttcctcca gtacaccttc caacgctaag aagcaagcga ctggaggtgc
     4441 tggcaaagct gctgccacca aagcaactgc tgccaaatcg gcagcctcta aagctcccca
     4501 gaatggcgca ggtgccaaga agaagggagg aaaggctgga ggacggaaga ggaaggatcc
     4561 accggtcatg gtgagcaagg gcgccgagct gttcaccggc atcgtgccca tcctgatcga
     4621 gctgaatggc gatgtgaatg gccacaagtt cagcgtgagc ggcgagggcg agggcgatgc
     4681 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ctgtgccctg
     4741 gcccaccctg gtgaccaccc tgagctacgg cgtgcagtgc ttctcacgct accccgatca
     4801 catgaagcag cacgacttct tcaagagcgc catgcctgag ggctacatcc aggagcgcac
     4861 catcttcttc gaggatgacg gcaactacaa gtcgcgcgcc gaggtgaagt tcgagggcga
     4921 taccctggtg aatcgcatcg agctgaccgg caccgatttc aaggaggatg gcaacatcct
     4981 gggcaataag atggagtaca actacaacgc ccacaatgtg tacatcatga ccgacaaggc
     5041 caagaatggc atcaaggtga acttcaagat ccgccacaac atcgaggatg gcagcgtgca
     5101 gctggccgac cactaccagc agaatacccc catcggcgat ggccctgtgc tgctgcccga
     5161 taaccactac ctgtccaccc agagcgccct gtccaaggac cccaacgaga agcgcgatca
     5221 catgatctac ttcggcttcg tgaccgccgc cgccatcacc cacggcatgg atgagctgta
     5281 caagtgagcg gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac
     5341 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     5401 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5461 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5521 atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     5581 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     5641 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     5701 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     5761 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     5821 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     5881 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     5941 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     6001 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat a
//
