LOCUS       sequence-155387-        7150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for constitutive or doxycycline-inducible expression of 
            H129N nuclease-dead mutant of human MRE11. 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 7150)
  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 7150)
  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-MRE11-H129N
FEATURES             Location/Qualifiers
     source          1..7150
                     /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(2231..2248)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2237..2461
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2507..2935
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2594..2613)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2804..2825
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2944..2964)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2949..3278
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3129..3264
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3330..3929
                     /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(3330..3349)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3826..3846
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4059..4180
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4096..4115)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4150..4169
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4230..4246)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4230..4246)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4243..4265)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4254..4270
                     /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(4278..4284,4285..4302,4303..4308))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4323..4344
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4461..4478)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4632..5217)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4712..4731)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5388..6179,6180..6248))
                     /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     6011..6030
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6249..6353)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6425..6804
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6805..7008
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6959..6979
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    7010..7028
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7031..7049
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     7059..7083
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 aagcttgcgg ccgccgccac catgtacccc tacgatgtgc ctgactacgc caccggtatg
       61 agtactgcag atgcacttga tgatgaaaac acatttaaaa tattagttgc aacagatatt
      121 catcttggat ttatggagaa agatgcagtc agaggaaatg atacgtttgt aacactcgat
      181 gaaattttaa gacttgccca ggaaaatgaa gtggatttta ttttgttagg tggtgatctt
      241 tttcatgaaa ataagccctc aaggaaaaca ttacatacct gcctcgagtt attaagaaaa
      301 tattgtatgg gtgatcggcc tgtccagttt gaaattctca gtgatcagtc agtcaacttt
      361 ggttttagta agtttccatg ggtgaactat caagatggca acctcaacat ttcaattcca
      421 gtgtttagta ttcatggcaa taatgacgat cccacagggg cagatgcact ttgtgccttg
      481 gacattttaa gttgtgctgg atttgtaaat cactttggac gttcaatgtc tgtggagaag
      541 atagacatta gtccggtttt gcttcaaaaa ggaagcacaa agattgcgct atatggttta
      601 ggatccattc cagatgaaag gctctatcga atgtttgtca ataaaaaagt aacaatgttg
      661 agaccaaagg aagatgagaa ctcttggttt aacttatttg tgattcatca gaacaggagt
      721 aaacatggaa gtactaactt cattccagaa caatttttgg atgacttcat tgatcttgtt
      781 atctggggcc atgaacatga gtgtaaaata gctccaacca aaaatgaaca acagctgttt
      841 tatatctcac aacctggaag ctcagtggtt acttctcttt ccccaggaga agctgtaaag
      901 aaacatgttg gtttgctgcg tattaaaggg aggaagatga atatgcataa aattcctctt
      961 cacacagtgc ggcagttttt catggaggat attgttctag ctaatcatcc agacattttt
     1021 aacccagata atcctaaagt aacccaagcc atacaaagct tctgtttgga gaagattgaa
     1081 gaaatgcttg aaaatgctga acgggaacgt ctgggtaatt ctcaccagcc agagaagcct
     1141 cttgtacgac tgcgagtgga ctatagtgga ggttttgaac ctttcagtgt tcttcgcttt
     1201 agccagaaat ttgtggatcg ggtagctaat ccaaaagaca ttatccattt tttcaggcat
     1261 agagaacaaa aggaaaaaac aggagaagag atcaactttg ggaaacttat cacaaagcct
     1321 tcagaaggaa caactttaag ggtagaagat cttgtaaaac agtactttca aaccgcagag
     1381 aagaatgtgc agctctcact gctaacagaa agagggatgg gtgaagcagt acaagaattt
     1441 gtggacaagg aggagaaaga tgccattgag gaattagtga aataccagtt ggaaaaaaca
     1501 cagcgatttc ttaaagaacg tcacattgat gccctcgaag acaaaatcga tgaggaggta
     1561 cgtcgtttca gagaaaccag acaaaaaaat actaatgaag aagatgatga agtccgtgag
     1621 gctatgacca gggccagagc actcagatct cagtcagagg agtctgcttc tgcctttagt
     1681 gctgatgacc ttatgagtat agatttagca gaacagatgg ctaatgactc tgatgatagc
     1741 atctcagcag caaccaacaa aggaagaggc cgaggaagag gtcgaagagg tggaagaggg
     1801 cagaattcag catcgagagg agggtctcaa agaggaagag cagacactgg tctggagact
     1861 tctacccgta gcaggaactc aaagactgct gtgtcagcat ctagaaatat gtctattata
     1921 gatgccttta aatctacaag acagcagcct tcccgaaatg tcactactaa gaattattca
     1981 gaggtgattg aggtagatga atcagatgtg gaagaagaca tttttcctac cacttcaaag
     2041 acagatcaaa ggtggtccag cacatcatcc agcaaaatca tgtcccagag tcaagtatcg
     2101 aaaggggttg attttgaatc aagtgaggat gatgatgatg atccttttat gaacactagt
     2161 tctttaagaa gaaatagaag ataaggtacc ctcgagacgc gtctgcagtc tagataaacc
     2221 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     2281 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     2341 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     2401 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     2461 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     2521 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2581 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2641 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2701 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2761 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2821 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2881 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     2941 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3001 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3061 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3121 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3181 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     3241 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     3301 ccattttcgg atctgatcag cacgtgacca tgaccgagta caagcccacg gtgcgcctcg
     3361 ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc
     3421 ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag
     3481 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     3541 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     3601 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     3661 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     3721 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     3781 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     3841 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     3901 ggtgcatgac ccgcaagccc ggtgcctgac acgtgctacg agatttcgat tccaccgccg
     3961 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     4021 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     4081 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4141 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctagccgtcg
     4201 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4261 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4321 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4381 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4441 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4501 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4561 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4621 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4681 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4741 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4801 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4861 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4921 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4981 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5041 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5101 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5161 ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5221 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5281 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5341 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5401 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5461 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5521 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5581 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5641 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5701 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5761 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5821 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5881 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5941 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6001 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6061 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6121 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6181 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6241 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6301 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6361 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc ctgtacgggc cagatatccg
     6421 cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     6481 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     6541 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     6601 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     6661 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     6721 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     6781 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     6841 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     6901 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     6961 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt
     7021 gatagagatc tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag
     7081 atcgcctgga gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc
     7141 agcctccggc
//
