LOCUS       sequence-155382-        7705 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for constitutive or doxy-inducible expression of N289A+H290A
            mutant of human CtIP 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 7705)
  AUTHORS   Chanut P, Britton S, Coates J, Jackson SP, Calsou P
  TITLE     Coordinated nuclease activities counteract Ku at single-ended DNA 
            double-strand breaks.
  JOURNAL   Nat Commun. 2016 Sep 19;7:12889. doi: 10.1038/ncomms12889.
  PUBMED    27641979
REFERENCE   2  (bases 1 to 7705)
  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: "2016-09-19"; volume: "7:12889. doi"; pages: " 
            10.1038/ncomms12889"
COMMENT     Sequence Label: pICE-HA-CtIP-siR-N289A-H290A
FEATURES             Location/Qualifiers
     source          1..7705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             25..51
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2786..2803)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2792..3016
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3062..3490
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3149..3168)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3359..3380
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3499..3519)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3504..3833
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3684..3819
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3746..3765
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3885..4484
                     /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(3885..3904)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4381..4401
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4614..4735
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4651..4670)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4705..4724
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4785..4801)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4785..4801)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4798..4820)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4809..4825
                     /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(4833..4839,4840..4857,4858..4863))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4878..4899
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5016..5033)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5187..5772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5267..5286)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5943..6734,6735..6803))
                     /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     6566..6585
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6804..6908)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6980..7359
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7360..7563
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7514..7534
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    7565..7583
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7586..7604
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     7614..7638
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 aagcttgcgg ccgccgccac catgtacccc tacgatgtgc ctgactacgc cggatccatg
       61 aacatctcgg gaagcagctg tggaagccct aactctgcag atacatctag tgactttaag
      121 gacctttgga caaaactaaa agaatgtcat gatagagaag tacaaggttt acaagtaaaa
      181 gtaaccaagt tgaagcagga acgaatctta gatgcacaaa gactagaaga attcttcacc
      241 aaaaatcaac agctgaggga acagcagaaa gtccttcatg aaaccattaa agttttagaa
      301 gatcggttaa gagcaggctt atgtgatcgc tgtgcagtaa ctgaagaaca tatgcggaaa
      361 aaacagcaag agtttgaaaa tatccggcag cagaatctta aacttattac agaacttatg
      421 aatgaaagga atactctaca ggaagaaaat aaaaagcttt ctgaacaact ccagcagaaa
      481 attgagaatg atcaacagca tcaagcagct gagcttgaat gtgaggaaga cgttattcca
      541 gattcaccga taacagcctt ctcattttct ggcgttaacc ggctacgaag aaaggagaac
      601 ccccatgtcc gatacataga acaaacacat actaaattgg agcactctgt gtgtgcaaat
      661 gaaatgagaa aagtttccaa gtcttcaact catccacaac ataatcctaa tgaaaatgaa
      721 attctagtag ctgacactta tgaccaaagt caatctccaa tggccaaagc acatggaaca
      781 agcagctata cccctgataa gtcatctttt aatttagcta cagttgttgc tgaaacactt
      841 ggacttggtg ttcaagaaga atctgaaact caaggtccca tgagccccct tggtgatgag
      901 ctctaccact gtctggaagg agccgccaag aaacagcctt ttgaggaatc tacaagaaat
      961 actgaagata gtttaagatt ttcagattct acttcaaaga ctcctcctca agaagaatta
     1021 cctactcgag tgtcatctcc tgtatttgga gctacctcta gtatcaaaag tggtttagat
     1081 ttgaatacaa gtttgtcccc ttctctttta cagcctggga aaaaaaaaca tctgaaaaca
     1141 ctccctttta gcaacacttg tatatctaga ttagaaaaaa ctagatcaaa atctgaagat
     1201 agtgcccttt tcacacatca cagtcttggg tctgaagtga acaagatcat tatccagtca
     1261 tctaataaac agatacttat aaataaaaat ataagtgaat ccctaggtga acagaatagg
     1321 actgagtacg gtaaagattc taacactgat aaacatttgg agcccctgaa atcattggga
     1381 ggccgaacat ccaaaaggaa gaaaactgag gaagaaagtg aacatgaagt aagctgcccc
     1441 caagcttctt ttgataaaga aaatgctttc ccttttccaa tggataatca gttttccatg
     1501 aatggagact gtgtgatgga taaacctctg gatctgtctg atcgattttc agctattcag
     1561 cgtcaagaga aaagccaagg aagtgagact tctaaaaaca aatttaggca agtgactctt
     1621 tatgaggctt tgaagaccat tccaaagggc ttttcctcaa gccgtaaggc ctcagatggc
     1681 aactgcacgt tgcccaaaga ttccccaggg gagccctgtt cacaggaatg catcatcctt
     1741 cagcccttga ataaatgctc tccagacaat aaaccatcat tacaaataaa agaagaaaat
     1801 gctgtcttta aaattcctct acgtccacgt gaaagtttgg agactgagaa tgttttagat
     1861 gacataaaga gtgctggttc tcatgagcca ataaaaatac aaaccaggtc agaccatgga
     1921 ggatgtgaac ttgcatcagt tcttcagtta aatccatgta gaactggtaa aataaagtct
     1981 ctacaaaaca accaagatgt atcctttgaa aatatccagt ggagtataga tccgggagca
     2041 gacctttctc agtataaaat ggatgttact gtaatagata caaaggatgg cagtcagtca
     2101 aaattaggag gagagacagt ggacatggac tgtacattgg ttagtgaaac cgttctctta
     2161 aaaatgaaga aacaagagca gaagggagaa aaaagttcaa atgaagaaag aaaaatgaat
     2221 gatagcttgg aagatatgtt tgatcggaca acacatgaag agtatgaatc ctgtttggca
     2281 gacagtttct cccaagcagc agatgaagag gaggaattgt ctactgccac aaagaaacta
     2341 cacactcatg gtgataaaca agacaaagtc aagcagaaag cgtttgtgga gccgtatttt
     2401 aaaggtgatg aaagagagac tagcttgcaa aattttcctc atattgaggt ggttcggaaa
     2461 aaagaggaga gaagaaaact gcttgggcac acgtgtaagg aatgtgaaat ttattatgca
     2521 gatatgccag cagaagaaag agaaaagaaa ttggcttcct gctcaagaca ccgattccgc
     2581 tacattccac ccaacacacc agagaatttt tgggaagttg gttttccttc cactcagact
     2641 tgtatggaaa gaggttatat taaggaagat cttgatcctt gtcctcgtcc aaaaagacgt
     2701 cagccttaca acgcaatatt ttctccaaaa ggcaaggagc agaagacata aacgcgtctg
     2761 cagtctagat aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2821 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2881 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2941 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     3001 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     3061 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3121 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3181 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3241 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3301 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3361 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3421 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3481 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3541 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3601 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3661 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3721 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3781 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3841 cgggagcttg tatatccatt ttcggatctg atcagcacgt gaccatgacc gagtacaagc
     3901 ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg
     3961 cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg
     4021 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     4081 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     4141 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     4201 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     4261 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     4321 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     4381 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     4441 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacacgtg ctacgagatt
     4501 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4561 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     4621 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4681 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4741 tctggctagc cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     4801 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4861 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4921 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4981 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     5041 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5101 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5161 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5221 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5281 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5341 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5401 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5461 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5521 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5581 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5641 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5701 aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5761 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5821 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5881 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5941 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6001 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6061 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6121 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6181 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6241 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6301 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6361 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6421 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6481 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6541 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6601 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6661 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6721 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6781 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6841 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6901 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcctgta
     6961 cgggccagat atccgcgttg acattgatta ttgactagtt attaatagta atcaattacg
     7021 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     7081 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     7141 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     7201 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     7261 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     7321 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     7381 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     7441 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     7501 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     7561 gctctcccta tcagtgatag agatctccct atcagtgata gagatcgtcg acgagctcgt
     7621 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
     7681 caccgggacc gatccagcct ccggc
//
