LOCUS       sequence-155730-        6844 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for constitutive or doxy-inducible expression of mutant 
            human Ku70 unable to bind DNA and resistant to siRNA. Confers 
            resistance to puro. 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-Mut6E
FEATURES             Location/Qualifiers
     source          1..6844
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        59..438
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        439..642
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     593..613
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    644..662
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    665..683
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     693..717
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             807..833
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2709..2726)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2715..2939
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2985..3413
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3072..3091)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3282..3303
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3422..3442)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3427..3756
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3607..3742
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3669..3688
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3808..4407
                     /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(3808..3827)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4304..4324
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4537..4658
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4574..4593)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4628..4647
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4708..4724)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4708..4724)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4721..4743)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4732..4748
                     /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(4756..4762,4763..4780,4781..4786))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4801..4822
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4939..4956)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5110..5695)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5190..5209)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5866..6657,6658..6726))
                     /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     6489..6508
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6727..6831)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaagtgccac ctgacgtcga cggatcggga gatcctgtac gggccagata tccgcgttga
       61 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      121 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      181 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      241 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      301 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      361 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      421 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      481 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      541 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      601 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctccctat cagtgataga
      661 gatctcccta tcagtgatag agatcgtcga cgagctcgtt tagtgaaccg tcagatcgcc
      721 tggagacgcc atccacgctg ttttgacctc catagaagac accgggaccg atccagcctc
      781 cggcaagctt accggtcgcc accatgtacc cctacgatgt gcctgactac gccctcgagt
      841 cagggtggga gtcatattac aaaaccgagg gcgatgaaga agcagaggaa gaacaagaag
      901 agaaccttga agcaagtgga gactataaat attcaggaag agatagtttg atttttttgg
      961 ttgatgcctc caaggctatg tttgaatctc aatccgagga cgaactcacc ccttttgaca
     1021 tgagcatcca gtgtatccaa agtgtgtaca tcagtaagat cataagcagt gatcgagatc
     1081 tcttggctgt ggtgttctat ggtaccgaga aagacaaaaa ttcagtgaat tttaaaaata
     1141 tttacgtctt acaggagctg gataatccag gtgcaaaacg aattctagag cttgaccagt
     1201 ttaaggggca gcagggacaa aaacgtttcc aagacatgat gggccacgga tctgactact
     1261 cactcagtga agtgctgtgg gtctgtgcca acctctttag tgatgtccaa ttcaagatga
     1321 gtcataagag gatcatgctg ttcaccaatg aagacaaccc ccatggcaat gacagtgcca
     1381 aagccagccg ggccaggacc aaagccggtg atctccgaga tacaggcatc ttccttgact
     1441 tgatgcacct gaagaaacct gggggctttg acatatcctt gttctacaga gatatcatca
     1501 gcatagcaga ggatgaggac ctcagggttc actttgagga atccagcaag ctagaagacc
     1561 tgttgcggaa ggttcgcgcc aaggagacaa ggaagcgagc actcagcagg ttaaagctga
     1621 agctcaacaa agatatagtg atctctgtgg gcatttataa tctggtccag aaggctctcg
     1681 aacctcctcc aatagagctc tatcgggaaa caaatgaacc agtgaaaacc aaggagcgga
     1741 cctttaatac aagtacaggc ggtttgcttc tgcctagcga taccaagagg tctcagatct
     1801 atgggagtcg tcagattata ctggaggaag aggaaacaga agagctagaa cggtttgatg
     1861 atccaggttt gatgctcatg ggtttcaagc cgttggtact gctgaagaaa caccattacc
     1921 tgaggccctc cctgttcgtg tacccagagg agtcgctggt gattgggagc tcaaccctgt
     1981 tcagtgctct gctcatcaag tgtctggaga aggaggttgc agcattgtgc agatacacac
     2041 ccgagaggaa catccctcct tattttgtgg ctttggtgcc acaggaagaa gagttggatg
     2101 accagaaaat tcaggtgact cctccaggct tccagctggt ctttttaccc tttgctgatg
     2161 ataaaaggaa gatgcccttt actgaaaaaa tcatggcaac tccagagcag gtgggcaaga
     2221 tgaaggctat cgttgagaag cttcgcttca catacagaag tgacagcttt gagaaccccg
     2281 tgctgcagca gcacttcagg aacctggagg ccttggcctt ggatttgatg gagccggaac
     2341 aagcagtgga cctgacattg cccaaggttg aagcaatgaa taaaagactg ggctccttgg
     2401 tggatgagtt taaggagctt gtttacccac cagattacaa tcctgaaggg aaagttacca
     2461 agagaaaaca cgataatgaa ggttctggaa gcaaaaggcc caaggtggag tattcagaag
     2521 aggagctgaa gacccacatc agcaagggta cgctgggcaa gttcactgtg cccatgctga
     2581 aagaggcctg ccgggcttac gggctgaaga gtgggctgaa gaagcaggag ctgctggaag
     2641 ccctcaccaa gcacttccag gactaagcgg ccgcacgcgt ctgcagtcta gataaacccg
     2701 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     2761 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     2821 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     2881 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     2941 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3001 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3061 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3121 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3181 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3241 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3301 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3361 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     3421 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3481 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     3541 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3601 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     3661 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     3721 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     3781 attttcggat ctgatcagca cgtgaccatg accgagtaca agcccacggt gcgcctcgcc
     3841 acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     3901 gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     3961 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     4021 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     4081 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     4141 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     4201 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     4261 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     4321 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     4381 tgcatgaccc gcaagcccgg tgcctgacac gtgctacgag atttcgattc caccgccgcc
     4441 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4501 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     4561 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4621 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct agccgtcgac
     4681 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     4741 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     4801 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     4861 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4921 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4981 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5041 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5101 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5161 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5221 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5281 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5341 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5401 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5461 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5521 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5581 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5641 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5701 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5761 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5821 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5881 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5941 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6001 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6061 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6121 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6181 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6241 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6301 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6361 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6421 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6481 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6541 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6601 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6661 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6721 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6781 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6841 ccga
//
