LOCUS       sequence-155729-        6844 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for constitutive or doxycycline-inducible expression of 
            wild-type human Ku70 resistant to a siRNA. Confers resistance to 
            puromycin. Use T-REx cells for doxycycline-inducible expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6844)
  AUTHORS   Lamaa A, Le Bras M, Skuli N, Britton S, Frit P, Calsou P, Prats H, 
            Cammas A, Millevoi S
  TITLE     A novel cytoprotective function for the DNA repair protein Ku in 
            regulating p53 mRNA translation and function.
  JOURNAL   EMBO Rep. 2016 Apr;17(4):508-18. doi: 10.15252/embr.201541181. Epub 
            2016 Mar 10.
  PUBMED    26964895
REFERENCE   2  (bases 1 to 6844)
  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"; doi: 
            "10.15252/embr.201541181"; journalName: "EMBO Rep"; date: "2016-04";
            volume: "17"; issue: "4"; pages: "508-18"
COMMENT     Sequence Label: pICE-HA-Ku70-siR-WT
FEATURES             Location/Qualifiers
     source          1..6844
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             22..48
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1924..1941)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1930..2154
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2200..2628
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2287..2306)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2497..2518
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2637..2657)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2642..2971
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2822..2957
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2884..2903
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3023..3622
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3023..3042)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3519..3539
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3752..3873
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3789..3808)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3843..3862
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3923..3939)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3923..3939)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3936..3958)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3947..3963
                     /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(3971..3977,3978..3995,3996..4001))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4016..4037
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4154..4171)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4325..4910)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4405..4424)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5081..5872,5873..5941))
                     /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     5704..5723
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5942..6046)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6118..6497
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6498..6701
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6652..6672
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    6703..6721
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6724..6742
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     6752..6776
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 agcttaccgg tcgccaccat gtacccctac gatgtgcctg actacgccct cgagtcaggg
       61 tgggagtcat attacaaaac cgagggcgat gaagaagcag aggaagaaca agaagagaac
      121 cttgaagcaa gtggagacta taaatattca ggaagagata gtttgatttt tttggttgat
      181 gcctccaagg ctatgtttga atctcaatcc gaggacgaac tcaccccttt tgacatgagc
      241 atccagtgta tccaaagtgt gtacatcagt aagatcataa gcagtgatcg agatctcttg
      301 gctgtggtgt tctatggtac cgagaaagac aaaaattcag tgaattttaa aaatatttac
      361 gtcttacagg agctggataa tccaggtgca aaacgaattc tagagcttga ccagtttaag
      421 gggcagcagg gacaaaaacg tttccaagac atgatgggcc acggatctga ctactcactc
      481 agtgaagtgc tgtgggtctg tgccaacctc tttagtgatg tccaattcaa gatgagtcat
      541 aagaggatca tgctgttcac caatgaagac aacccccatg gcaatgacag tgccaaagcc
      601 agccgggcca ggaccaaagc cggtgatctc cgagatacag gcatcttcct tgacttgatg
      661 cacctgaaga aacctggggg ctttgacata tccttgttct acagagatat catcagcata
      721 gcagaggatg aggacctcag ggttcacttt gaggaatcca gcaagctaga agacctgttg
      781 cggaaggttc gcgccaagga gacaaggaag cgagcactca gcaggttaaa gctgaagctc
      841 aacaaagata tagtgatctc tgtgggcatt tataatctgg tccagaaggc tctcaagcct
      901 cctccaataa agctctatcg ggaaacaaat gaaccagtga aaaccaagac ccggaccttt
      961 aatacaagta caggcggttt gcttctgcct agcgatacca agaggtctca gatctatggg
     1021 agtcgtcaga ttatactgga gaaagaggaa acagaagagc taaaacggtt tgatgatcca
     1081 ggtttgatgc tcatgggttt caagccgttg gtactgctga agaaacacca ttacctgagg
     1141 ccctccctgt tcgtgtaccc agaggagtcg ctggtgattg ggagctcaac cctgttcagt
     1201 gctctgctca tcaagtgtct ggagaaggag gttgcagcat tgtgcagata cacaccccgc
     1261 aggaacatcc ctccttattt tgtggctttg gtgccacagg aagaagagtt ggatgaccag
     1321 aaaattcagg tgactcctcc aggcttccag ctggtctttt taccctttgc tgatgataaa
     1381 aggaagatgc cctttactga aaaaatcatg gcaactccag agcaggtggg caagatgaag
     1441 gctatcgttg agaagcttcg cttcacatac agaagtgaca gctttgagaa ccccgtgctg
     1501 cagcagcact tcaggaacct ggaggccttg gccttggatt tgatggagcc ggaacaagca
     1561 gtggacctga cattgcccaa ggttgaagca atgaataaaa gactgggctc cttggtggat
     1621 gagtttaagg agcttgttta cccaccagat tacaatcctg aagggaaagt taccaagaga
     1681 aaacacgata atgaaggttc tggaagcaaa aggcccaagg tggagtattc agaagaggag
     1741 ctgaagaccc acatcagcaa gggtacgctg ggcaagttca ctgtgcccat gctgaaagag
     1801 gcctgccggg cttacgggct gaagagtggg ctgaagaagc aggagctgct ggaagccctc
     1861 accaagcact tccaggacta agcggccgca cgcgtctgca gtctagataa acccgctgat
     1921 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     1981 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     2041 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2101 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     2161 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     2221 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2281 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2341 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     2401 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     2461 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     2521 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     2581 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     2641 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2701 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2761 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2821 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2881 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     2941 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     3001 cggatctgat cagcacgtga ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     3061 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     3121 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     3181 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     3241 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     3301 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     3361 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     3421 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     3481 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     3541 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     3601 gacccgcaag cccggtgcct gacacgtgct acgagatttc gattccaccg ccgccttcta
     3661 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     3721 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     3781 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     3841 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggctagccg tcgacctcta
     3901 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3961 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     4021 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4081 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4141 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4201 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4261 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4321 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4381 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4441 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4501 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4561 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4621 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4681 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4741 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4801 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4861 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4921 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4981 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5041 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5101 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5161 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5221 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5281 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5341 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5401 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5461 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5521 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5581 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5641 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5701 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5761 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5821 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5881 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5941 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6001 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6061 aagtgccacc tgacgtcgac ggatcgggag atcctgtacg ggccagatat ccgcgttgac
     6121 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     6181 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     6241 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     6301 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     6361 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     6421 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     6481 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     6541 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     6601 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     6661 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctccctatc agtgatagag
     6721 atctccctat cagtgataga gatcgtcgac gagctcgttt agtgaaccgt cagatcgcct
     6781 ggagacgcca tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc
     6841 ggca
//
