LOCUS       sequence-178890-        7366 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To establish stable line expressing Dsup in constitutive manner.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7366)
  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 7366)
  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: pCXN2KS-Dsup
FEATURES             Location/Qualifiers
     source          1..7366
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             286..1089
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(349..368)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     959..978
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1096..1144
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        1154..1533
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1535..1810
                     /label=chicken beta-actin promoter
     intron          1811..2822
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2746..2768
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2830..2849
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     2882..2901
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2882..2900
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4282..4302)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4282..4300)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4388..4407)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4453..4508
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4507..4526)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4869..4885)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4869..4885)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4882..4904)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4893..4909
                     /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)."
     promoter        complement(join(4917..4923,4924..4941,4942..4947))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4962..4983
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5042..5237
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5088..5223
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5243..5377
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5280..5299)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5334..5353
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5445..5462)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5616..6204)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5696..5715)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6375..7166,7167..7235))
                     /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     6998..7017
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7236..7340)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgagcag tgtggttttc aagaggaagc aaaaagcctc tccacccagg cctggaatgt
       61 ttccacccaa tgtcgagcag tgtggttttg caagaggaag caaaaagcct ctccacccag
      121 gcctggaatg tttccaccca atgtcgagca aaccccgccc agcgtcttgt cattggcgaa
      181 ttaattcgaa cacgcagatg cagtcggggc ggcgcggtcc caggtccact tcgcatatta
      241 aggtgacgcg tgtggcctcg aacaccgagc gaccctgcag ccaatatggg atcggccatt
      301 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
      361 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
      421 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
      481 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
      541 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
      601 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
      661 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
      721 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
      781 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgat
      841 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
      901 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
      961 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     1021 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     1081 ttcttctgag gggatcggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt
     1141 gttcggatcc gtcgacattg attattgact agttattaat agtaatcaat tacggggtca
     1201 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     1261 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     1321 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     1381 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     1441 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     1501 tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc
     1561 actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta
     1621 ttttgtgcag cgatgggggc gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg
     1681 cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct
     1741 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc
     1801 gcggcgggcg ggagtcgctg cgttgccttc gccccgtgcc ccgctccgcg ccgcctcgcg
     1861 ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc
     1921 ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg ctcgtttctt ttctgtggct
     1981 gcgtgaaagc cttaaagggc tccgggaggg ccctttgtgc gggggggagc ggctcggggg
     2041 gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg cccgcgctgc ccggcggctg
     2101 tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc gtgtgcgcga ggggagcgcg
     2161 gccgggggcg gtgccccgcg gtgcgggggg gctgcgaggg gaacaaaggc tgcgtgcggg
     2221 gtgtgtgcgt gggggggtga gcagggggtg tgggcgcggc ggtcgggctg taaccccccc
     2281 ctgcaccccc ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtgcg
     2341 gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc
     2401 ggggcggggc cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg ccccggagcg
     2461 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
     2521 gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc
     2581 gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg
     2641 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct
     2701 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
     2761 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
     2821 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattcgcgc
     2881 gtaatacgac tcactatagg gcgaattggg tacatggcat ccacacacca atcatccaca
     2941 gaaccctctt ccacaggtaa atctgaggaa acgaagaaag atgcttcgca agggagcggg
     3001 caagactcca agaacgtaac cgttaccaaa ggtaccggtt cctccgccac ctcagctgcc
     3061 attgtcaaga caggaggatc ccaaggcaaa gattcctcta ctacagcggg ctcttctagt
     3121 actcagggac agaagttcag tactacacct accgacccga aaactttcag ctctgaccaa
     3181 aaggagaaat ccaaaagccc agccaaagaa gtcccgtctg gtggcgatag taagtcccaa
     3241 ggtgacacca agtctcaaag cgacgccaaa tcttctggac aaagtcaggg ccagtctaaa
     3301 gacagcggca aatcatcttc cgacagtagc aagagtcact ctgtcatcgg agctgtcaaa
     3361 gacgtcgttg caggcgccaa agatgtcgca ggaaaagccg tcgaggatgc tcctagcatc
     3421 atgcatactg cagtcgatgc tgtgaagaac gcagccacga ctgtgaagga tgtggcatcg
     3481 tcggctgcat cgactgtggc ggagaaggta gtcgatgctt accacagtgt ggtgggagac
     3541 aagacggacg acaagaaaga gggcgagcac agcggcgaca agaaggacga ctccaaagct
     3601 ggaagtggct ctggacaagg tggtgacaac aagaagtctg aaggagagac ttctggccaa
     3661 gcagaatcca gctctggcaa cgaaggagct gctccagcca aaggccgtgg tcgtggacgg
     3721 cctccagcag ctgctaaagg agttgctaag ggtgctgcaa agggcgctgc cgcctccaaa
     3781 ggagccaaga gcggtgctga atcctccaag ggaggagaac agtcgtcagg agatatcgag
     3841 atggcagatg cttcctccaa gggaggctcg gaccagaggg attccgcggc gaccgttggc
     3901 gaaggtggtg catcaggcag tgagggtgga gctaagaaag gcagagggcg gggcgctggt
     3961 aagaaagcgg atgcgggtga tacgtccgct gagccgcctc ggcggtcgtc ccgcctgacg
     4021 tcttcaggta caggggcggg ttccgctcca gctgcagcga aaggcggagc gaagcgtgct
     4081 gcttcttcct ccagtacacc ttccaacgct aagaagcaag cgactggagg tgctggcaaa
     4141 gctgctgcca ccaaagcaac tgctgccaaa tcggcagcct ctaaagctcc ccagaatggc
     4201 gcaggtgcca agaagaaggg aggaaaggct ggaggacgga agaggaagta agccaccgcg
     4261 gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgaattc actcctcagg
     4321 tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc
     4381 actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc
     4441 tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt
     4501 gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt
     4561 tggtttagag tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat
     4621 aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag
     4681 ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat
     4741 ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc
     4801 cagtcatagc tgtccctctt ctcttatgga gatccctcga cctgcagccc aagcttggcg
     4861 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4921 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4981 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc
     5041 cgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     5101 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     5161 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5221 gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag
     5281 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5341 gtccaaactc atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa
     5401 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5461 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5521 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5581 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5641 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5701 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5761 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5821 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5881 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5941 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6001 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     6061 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6121 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6181 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6241 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6301 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6361 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6421 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6481 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6541 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6601 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     6661 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     6721 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6781 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6841 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6901 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6961 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7021 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7081 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7141 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7201 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7261 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7321 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctg
//
