LOCUS       sequence-412378-        8176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To generate stable cell line expressing Halo-human MRE11(H129N) by 
            Retroviral infection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8176)
  AUTHORS   Cho MG, Kumar RJ, Lin CC, Boyer JA, Shahir JA, Fagan-Solis K, 
            Simpson DA, Fan C, Foster CE, Goddard AM, Lerner LM, Ellington SW, 
            Wang Q, Wang Y, Ho AY, Liu P, Perou CM, Zhang Q, McGinty RK, Purvis 
            JE, Gupta GP
  TITLE     MRE11 liberates cGAS from nucleosome sequestration during 
            tumorigenesis.
  JOURNAL   Nature. 2024 Jan 10. doi: 10.1038/s41586-023-06889-6.
  PUBMED    38200309
REFERENCE   2  (bases 1 to 8176)
  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: "Nature. 
            2024 Jan 10. doi: 10.1038/s41586-023-06889-6."
COMMENT     Sequence Label: pBABE-puro-Halo-hMRE11(H129N)
FEATURES             Location/Qualifiers
     source          1..8176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      262..850
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     751..770
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1004..1021
                     /label=L4440
                     /note="L4440 vector, forward primer"
     LTR             1151..1620
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(1841..2440)
                     /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(1924..1944)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2421..2440
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(2450..2779)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2464..2599
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2764..2784
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(4968..4988)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             complement(5001..5891)
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     regulatory      5888..5897
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(5932..6348)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(5948..5964)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(5980..6002)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    6549..6748
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             6813..7282
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        7313..7417
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7418..7486,7487..8176,1..102)
                     /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     complement(7636..7655)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
       61 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
      121 tcatatatac tttagattga tttgcggccg caaaaggatc taggtgaaga tcctttttga
      181 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
      241 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
      301 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
      361 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
      421 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
      481 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
      541 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
      601 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
      661 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
      721 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
      781 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
      841 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
      901 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
      961 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     1021 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     1081 atgcaactat ggccatttaa tgtaaatact taagaaaaaa aaccaaatta attttgatac
     1141 atgctgcatg tgaaagaccc ccgctgacgg gtagtcaatc actcagagga gaccctccca
     1201 aggaacagcg agaccacaag tcggatgcaa ctgcaagagg gtttattgga tacacgggta
     1261 cccgggcgac tcagtcaatc ggaggactgg cgccccgagt gaggggttgt gggctctttt
     1321 attgagctcg gggagcagaa gcgcgcgaac agaagcgaga agcgaactga ttggttagtt
     1381 caaataaggc acagggtcat ttcaggtcct tggggcaccc tggaaacatc tgatggttct
     1441 ctagaaactg ctgagggctg gaccgcatct ggggaccatc tgttcttggc cctgagccgg
     1501 ggcaggaact gcttaccaca gatatcctgt ttggcccata ttcagctgtt ccatctgttc
     1561 ttggccctga gccggggcag gaactgctta ccacagatat cctgtttggc ccatattcag
     1621 ctgttccatc tgttcctgac cttgatctga acttctctat tctcagttat gtatttttcc
     1681 atgccttgca aaatggcgtt acttaagcta gcttgccaaa cctacaggtg gggtctttca
     1741 ttcccccctt tttctggaga ctaaataaaa tcttttattt tatcgatgca tggggtcgtg
     1801 cgctcctttc ggtcgggcgc tgcgggtcgt ggggcgggcg tcaggcaccg ggcttgcggg
     1861 tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga
     1921 gccgctcgta gaaggggagg ttgcggggcg cggaggtctc caggaaggcg ggcaccccgg
     1981 cgcgctcggc cgcctccact ccggggagca cgacggcgct gcccagaccc ttgccctggt
     2041 ggtcgggcga gacgccgacg gtggccagga accacgcggg ctccttgggc cggtgcggcg
     2101 ccaggaggcc ttccatctgt tgctgcgcgg ccagccggga accgctcaac tcggccatgc
     2161 gcgggccgat ctcggcgaac accgcccccg cttcgacgct ctccggcgtg gtccagaccg
     2221 ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga
     2281 ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg gtccggatcg acggtgtggc
     2341 gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg tacggccctg gggacgtcgt
     2401 cgcgggtggc gaggcgcacc gtgggcttgt actcggtcat ggtaagcttt ttgcaaaagc
     2461 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct
     2521 cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg
     2581 gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga
     2641 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg
     2701 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     2761 cacaccctaa ctgacacaca ttccacaggg tcgactgcag aattcgaacc ttatcttcta
     2821 tttcttctta aagaactagt gttcataaaa ggatcatcat catcatcctc acttgattca
     2881 aaatcaaccc ctttcgatac ttgactctgg gacatgattt tgctggatga tgtgctggac
     2941 cacctttgat ctgtctttga agtggtagga aaaatgtctt cttccacatc tgattcatct
     3001 acctcaatca cctctgaata attcttagta gtgacatttc gggaaggctg ctgtcttgta
     3061 gatttaaagg catctataat agacatattt ctagatgctg acacagcagt ctttgagttc
     3121 ctgctacggg tagaagtctc cagaccagtg tctgctcttc ctctttgaga ccctcctctc
     3181 gatgctgaat tctgccctct tccacctctt cgacctcttc ctcggcctct tcctttgttg
     3241 gttgctgctg agatgctatc atcagagtca ttagccatct gttctgctaa atctatactc
     3301 ataaggtcat cagcactaaa ggcagaagca gactcctctg actgagatct gagtgctctg
     3361 gccctggtca tagcctcacg gacttcatca tcttcttcat tagtattttt ttgtctggtt
     3421 tctctgaaac gacgtacctc ctcatcgatt ttgtcttcga gggcatcaat gtgacgttct
     3481 ttaagaaatc gctgtgtttt ttccaactgg tatttcacta attcctcaat ggcatctttc
     3541 tcctccttgt ccacaaattc ttgtactgct tcacccatcc ctctttctgt tagcagtgag
     3601 agctgcacat tcttctctgc ggtttgaaag tactgtttta caagatcttc tacccttaaa
     3661 gttgttcctt ctgaaggctt tgtgataagt ttcccaaagt tgatctcttc tcctgttttt
     3721 tccttttgtt ctctatgcct gaaaaaatgg ataatgtctt ttggattagc tacccgatcc
     3781 acaaatttct ggctaaagcg aagaacactg aaaggttcaa aacctccact atagtccact
     3841 cgcagtcgta caagaggctt ctctggctgg tgagaattac ccagacgttc ccgttcagca
     3901 ttttcaagca tttcttcaat cttctccaaa cagaagcttt gtatggcttg ggttacttta
     3961 ggattatctg ggttaaaaat gtctggatga ttagctagaa caatatcctc catgaaaaac
     4021 tgccgcactg tgtgaagagg aattttatgc atattcatct tcctcccttt aatacgcagc
     4081 aaaccaacat gtttctttac agcttctcct ggggaaagag aagtaaccac tgagcttcca
     4141 ggttgtgaga tataaaacag ctgttgttca tttttggttg gagctatttt acactcatgt
     4201 tcatggcccc agataacaag atcaatgaag tcatccaaaa attgttctgg aatgaagtta
     4261 gtacttccat gtttactcct gttctgatga atcacaaata agttaaacca agagttctca
     4321 tcttcctttg gtctcaacat tgttactttt ttattgacaa acattcgata gagcctttca
     4381 tctggaatgg atcctaaacc atatagcgca atctttgtgc ttcctttttg aagcaaaacc
     4441 ggactaatgt ctatcttctc cacagacatt gaacgtccaa agtgatttac aaatccagca
     4501 caacttaaaa tgtccaaggc acaaagtgca tctgcccctg tgggatcgtc attattgcca
     4561 tgaatactaa acactggaat tgaaatgttg aggttgccat cttgatagtt cacccatgga
     4621 aacttactaa aaccaaagtt gactgactga tcactgagaa tttcaaactg gacaggccga
     4681 tcacccatac aatattttct taataactcg aggcaggtat gtaatgtttt ccttgagggc
     4741 ttattttcat gaaaaagatc accacctaac aaaataaaat ccacttcatt ttcctgggca
     4801 agtcttaaaa tttcatcgag tgttacaaac gtatcatttc ctctgactgc atctttctcc
     4861 ataaatccaa gatgaatatc tgttgcaact aatattttaa atgtgttttc atcatcaagt
     4921 gcatctgcag tactcatctt cgaagcttga gctcgagatc tgttatcgct ctgaaagtac
     4981 agatcctcag tggttggctc accggaaatc tccagagtag acagccagcg cgcgatctcg
     5041 ctgccgatca ggtccgggtt gtcttcttgc agcagattca gacccgggcc gatgtccaca
     5101 gccttgcagt taggcaggct tttggccagg cgagcggctt cggccggtgg gatcagaacg
     5161 cctggggtgc cccagaacag cagcttcggg acaggggact ggtgcagcca gtccatgtat
     5221 tcttcgacca gcgcgacgat gttcgctggc tcaccggcga ttggcagctc gtttgggaag
     5281 cgccacagtg gctcgcggtc aacaggattc aggaacggct cgcggtaatg gtccatctcg
     5341 acttcagtca gcgggcggac gacacccatc ggcagcgtac cctcgataaa aacgttctga
     5401 tcgatgatca gcttgcggcc gacgtcggtg gtgcggaagg cctggaaggt ctcgcgggca
     5461 aattctggcc attcgtccca ggtcgggata gggcggatga actccataaa tgcaatacct
     5521 ttgacgcgct ctggattgcg cttggcccag tggaaaccca gagcggagcc ccagtcgtga
     5581 atgaccagga cgacctcttc cagacccagg gcttcgatga aggcatccat gaagcggacg
     5641 tggtcgtcga agaaataacc caggtctggt ttgtcggatt tgcccatacc gatcaggtct
     5701 ggagcaatgc agcgatgggt cggtgcaaca tgcgggatga tgttgcgcca cacgtaggag
     5761 gaggtcgggt taccgtgcag gaacagcaca ggggtgccat cgcgcggacc aacatcgacg
     5821 tagtgcatgc gctcgcccag gacttccaca taatgggggt cgaatggaaa gccagtaccg
     5881 atttctgcca tggtggcgac cggtagcgta ccaccacact gggatccggc cggcgcctag
     5941 agaaggagtg agggctggat aaagggagga tcgaggcggg gtcgaacgag gaggttcaag
     6001 ggggagagac ggggcggatg gaggaagagg aggcggaggc ttagggtgta caaagggctt
     6061 gacccaggga ggggggtcaa aagccaaggc ttcccaggtc acgatgtagg ggacctggtc
     6121 tgggtgtcca tgcgggccag gtgaaaagac cttgatctta acctgggtga tgaggtctcg
     6181 gttaaaggtg ccgtctcgcg gccatccgac gttaaaggtt ggccattctg cagagcagaa
     6241 ggtaacccaa cgtctcttct tgacatctac cgactggttg tgagcgatcc gctcgacatc
     6301 tttccagtga cctaaggtca aacttaaggg agtggtaaca gtctggccct aattttcaga
     6361 caaatacaga aacacagtca gacagagaca acacagaacg atgctgcagc gctgcagcag
     6421 acaagacgcg cggcttcggt tccaaaccga aagcaaaaat tcagacggag gcgggaactg
     6481 ttttaggttc tcgtctccta ccagaaccac atatcctgac ggggtcggat tccacatcga
     6541 ctcccttcct caggtcgggc cacaaaaacg gcccccaaag tccctgggac gtctcccagg
     6601 gttgcggccg ggtgttcaga actcgtcagt tccaccacgg gtccgccaga tacagagcta
     6661 gttagctaac tagtaccgac gcaggcgcat aaaatcagtc atagacacta gacaatcgga
     6721 cagacacaga taagttgctg gccagcttgc ctcccggtgg tgggtcggtg gtccctgggc
     6781 aggggtctcc cgatcccgga cgagccccca aatgaaagac ccccgctgac gggtagtcaa
     6841 tcactcagag gagaccctcc caaggaacag cgagaccaca agtcggatgc aactgcaaga
     6901 gggtttattg gatacacggg tacccgggcg actcagtcaa tcggaggact ggcgccccga
     6961 gtgaggggtt gtgggctctt ttattgagct cggggagcag aagcgcgcga acagaagcga
     7021 gaagcgaact gattggttag ttcaaataag gcacagggtc atttcaggtc cttggggcac
     7081 cctggaaaca tctgatggtt ctctagaaac tgctgagggc tggaccgcat ctggggacca
     7141 tctgttcttg gccctgagcc ggggcaggaa ctgcttacca cagatatcct gtttggccca
     7201 tattcagctg ttccatctgt tcttggccct gagccggggc aggaactgct taccacagat
     7261 atcctgtttg gcccatattc aggctgcagg tggcactttt cggggaaatg tgcgcggaac
     7321 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     7381 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     7441 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     7501 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     7561 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     7621 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     7681 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     7741 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     7801 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     7861 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     7921 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     7981 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     8041 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     8101 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     8161 gccagatggt aagccc
//
