LOCUS       sequence-412499-        7894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To generate stable cell line expressing Halo-human 
            MRE11(DB1-Delta/DB2-Delta) by Retroviral infection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7894)
  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 7894)
  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(DB1-Delta/DB2-Delta)
FEATURES             Location/Qualifiers
     source          1..7894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(105..124)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      907..1495
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1396..1415
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1649..1666
                     /label=L4440
                     /note="L4440 vector, forward primer"
     LTR             1796..2265
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(2486..3085)
                     /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(2569..2589)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3066..3085
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(3095..3424)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3109..3244
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3409..3429
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(5331..5351)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             complement(5364..6254)
                     /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      6251..6260
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(6295..6711)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(6311..6327)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(6343..6365)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    6912..7111
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             7176..7645
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        7676..7780
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7781..7849,7850..7894,1..747)
                     /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"
ORIGIN
        1 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
       61 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
      121 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
      181 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
      241 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
      301 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
      361 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
      421 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
      481 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
      541 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
      601 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
      661 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
      721 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
      781 attgatttgc ggccgcaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
      841 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
      901 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
      961 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     1021 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     1081 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     1141 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     1201 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     1261 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     1321 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     1381 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     1441 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     1501 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     1561 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     1621 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     1681 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca actatggcca
     1741 tttaatgtaa atacttaaga aaaaaaacca aattaatttt gatacatgct gcatgtgaaa
     1801 gacccccgct gacgggtagt caatcactca gaggagaccc tcccaaggaa cagcgagacc
     1861 acaagtcgga tgcaactgca agagggttta ttggatacac gggtacccgg gcgactcagt
     1921 caatcggagg actggcgccc cgagtgaggg gttgtgggct cttttattga gctcggggag
     1981 cagaagcgcg cgaacagaag cgagaagcga actgattggt tagttcaaat aaggcacagg
     2041 gtcatttcag gtccttgggg caccctggaa acatctgatg gttctctaga aactgctgag
     2101 ggctggaccg catctgggga ccatctgttc ttggccctga gccggggcag gaactgctta
     2161 ccacagatat cctgtttggc ccatattcag ctgttccatc tgttcttggc cctgagccgg
     2221 ggcaggaact gcttaccaca gatatcctgt ttggcccata ttcagctgtt ccatctgttc
     2281 ctgaccttga tctgaacttc tctattctca gttatgtatt tttccatgcc ttgcaaaatg
     2341 gcgttactta agctagcttg ccaaacctac aggtggggtc tttcattccc ccctttttct
     2401 ggagactaaa taaaatcttt tattttatcg atgcatgggg tcgtgcgctc ctttcggtcg
     2461 ggcgctgcgg gtcgtggggc gggcgtcagg caccgggctt gcgggtcatg caccaggtgc
     2521 gcggtccttc gggcacctcg acgtcggcgg tgacggtgaa gccgagccgc tcgtagaagg
     2581 ggaggttgcg gggcgcggag gtctccagga aggcgggcac cccggcgcgc tcggccgcct
     2641 ccactccggg gagcacgacg gcgctgccca gacccttgcc ctggtggtcg ggcgagacgc
     2701 cgacggtggc caggaaccac gcgggctcct tgggccggtg cggcgccagg aggccttcca
     2761 tctgttgctg cgcggccagc cgggaaccgc tcaactcggc catgcgcggg ccgatctcgg
     2821 cgaacaccgc ccccgcttcg acgctctccg gcgtggtcca gaccgccacc gcggcgccgt
     2881 cgtccgcgac ccacaccttg ccgatgtcga gcccgacgcg cgtgaggaag agttcttgca
     2941 gctcggtgac ccgctcgatg tggcggtccg gatcgacggt gtggcgcgtg gcggggtagt
     3001 cggcgaacgc ggcggcgagg gtgcgtacgg ccctggggac gtcgtcgcgg gtggcgaggc
     3061 gcaccgtggg cttgtactcg gtcatggtaa gctttttgca aaagcctagg cctccaaaaa
     3121 agcctcctca ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa
     3181 taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg
     3241 ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca
     3301 tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac taattgagat
     3361 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac
     3421 acacattcca cagggtcgac tgcagaattc gaaccttagg aaggctgctg tcttgtagat
     3481 ttaaaggcat ctataataga catatttcta gatgctgaca cagcagtctt tgagttcctg
     3541 ctacgggtag aagtctccag accagtgtct gctcttcctc tttgagaccc tcctctcgat
     3601 gctgaattct gccctcttcc acctcttcga cctcttcctc ggcctcttcc tttgttggtt
     3661 gctgctgaga tgctatcatc agagtcatta gccatctgtt ctgctaaatc tatactcata
     3721 aggtcatcag cactaaaggc agaagcagac tcctctgact gagatctgag tgctctggcc
     3781 ctggtcatag cctcacggac ttcatcatct tcttcattag tatttttttg tctggtttct
     3841 ctgaaacgac gtacctcctc atcgattttg tcttcgaggg catcaatgtg acgttcttta
     3901 agaaatcgct gtgttttttc caactggtat ttcactaatt cctcaatggc atctttctcc
     3961 tccttgtcca caaattcttg tactgcttca cccatccctc tttctgttag cagtgagagc
     4021 tgcacattct tctctgcggt ttgaaagtac tgttttacaa gatcttctac ccttaaagtt
     4081 gttccttctt gttctctatg cctgaaaaaa tggataatgt cttttggatt agctacccga
     4141 tccacaaatt tctggctaaa gcgaagaaca ctgaaaggtt caaaacctcc actatagtcc
     4201 actcgcagtc gtacaagagg cttctctggc tggtgagaat tacccagacg ttcccgttca
     4261 gcattttcaa gcatttcttc aatcttctcc aaacagaagc tttgtatggc ttgggttact
     4321 ttaggattat ctgggttaaa aatgtctgga tgattagcta gaacaatatc ctccatgaaa
     4381 aactgccgca ctgtgtgaag aggaatttta tgcatattca tcttcctccc tttaatacgc
     4441 agcaaaccaa catgtttctt tacagcttct cctggggaaa gagaagtaac cactgagctt
     4501 ccaggttgtg agatataaaa cagctgttgt tcatttttgg ttggagctat tttacactca
     4561 tgttcatggc cccagataac aagatcaatg aagtcatcca aaaattgttc tggaatgaag
     4621 ttagtacttc catgtttact cctgttctga tgaatcacaa ataagttaaa ccaagagttc
     4681 tcatcttcct ttggtctcaa cattgttact tttttattga caaacattcg atagagcctt
     4741 tcatctggaa tggatcctaa accatatagc gcaatctttg tgcttccttt ttgaagcaaa
     4801 accggactaa tgtctatctt ctccacagac attgaacgtc caaagtgatt tacaaatcca
     4861 gcacaactta aaatgtccaa ggcacaaagt gcatctgccc ctgtgggatc gtcatgattg
     4921 ccatgaatac taaacactgg aattgaaatg ttgaggttgc catcttgata gttcacccat
     4981 ggaaacttac taaaaccaaa gttgactgac tgatcactga gaatttcaaa ctggacaggc
     5041 cgatcaccca tacaatattt tcttaataac tcgaggcagg tatgtaatgt tttccttgag
     5101 ggcttatttt catgaaaaag atcaccacct aacaaaataa aatccacttc attttcctgg
     5161 gcaagtctta aaatttcatc gagtgttaca aacgtatcat ttcctctgac tgcatctttc
     5221 tccataaatc caagatgaat atctgttgca actaatattt taaatgtgtt ttcatcatca
     5281 agtgcatctg cagtactcat cttcgaagct tgagctcgag atctgttatc gctctgaaag
     5341 tacagatcct cagtggttgg ctcaccggaa atctccagag tagacagcca gcgcgcgatc
     5401 tcgctgccga tcaggtccgg gttgtcttct tgcagcagat tcagacccgg gccgatgtcc
     5461 acagccttgc agttaggcag gcttttggcc aggcgagcgg cttcggccgg tgggatcaga
     5521 acgcctgggg tgccccagaa cagcagcttc gggacagggg actggtgcag ccagtccatg
     5581 tattcttcga ccagcgcgac gatgttcgct ggctcaccgg cgattggcag ctcgtttggg
     5641 aagcgccaca gtggctcgcg gtcaacagga ttcaggaacg gctcgcggta atggtccatc
     5701 tcgacttcag tcagcgggcg gacgacaccc atcggcagcg taccctcgat aaaaacgttc
     5761 tgatcgatga tcagcttgcg gccgacgtcg gtggtgcgga aggcctggaa ggtctcgcgg
     5821 gcaaattctg gccattcgtc ccaggtcggg atagggcgga tgaactccat aaatgcaata
     5881 cctttgacgc gctctggatt gcgcttggcc cagtggaaac ccagagcgga gccccagtcg
     5941 tgaatgacca ggacgacctc ttccagaccc agggcttcga tgaaggcatc catgaagcgg
     6001 acgtggtcgt cgaagaaata acccaggtct ggtttgtcgg atttgcccat accgatcagg
     6061 tctggagcaa tgcagcgatg ggtcggtgca acatgcggga tgatgttgcg ccacacgtag
     6121 gaggaggtcg ggttaccgtg caggaacagc acaggggtgc catcgcgcgg accaacatcg
     6181 acgtagtgca tgcgctcgcc caggacttcc acataatggg ggtcgaatgg aaagccagta
     6241 ccgatttctg ccatggtggc gaccggtagc gtaccaccac actgggatcc ggccggcgcc
     6301 tagagaagga gtgagggctg gataaaggga ggatcgaggc ggggtcgaac gaggaggttc
     6361 aagggggaga gacggggcgg atggaggaag aggaggcgga ggcttagggt gtacaaaggg
     6421 cttgacccag ggaggggggt caaaagccaa ggcttcccag gtcacgatgt aggggacctg
     6481 gtctgggtgt ccatgcgggc caggtgaaaa gaccttgatc ttaacctggg tgatgaggtc
     6541 tcggttaaag gtgccgtctc gcggccatcc gacgttaaag gttggccatt ctgcagagca
     6601 gaaggtaacc caacgtctct tcttgacatc taccgactgg ttgtgagcga tccgctcgac
     6661 atctttccag tgacctaagg tcaaacttaa gggagtggta acagtctggc cctaattttc
     6721 agacaaatac agaaacacag tcagacagag acaacacaga acgatgctgc agcgctgcag
     6781 cagacaagac gcgcggcttc ggttccaaac cgaaagcaaa aattcagacg gaggcgggaa
     6841 ctgttttagg ttctcgtctc ctaccagaac cacatatcct gacggggtcg gattccacat
     6901 cgactccctt cctcaggtcg ggccacaaaa acggccccca aagtccctgg gacgtctccc
     6961 agggttgcgg ccgggtgttc agaactcgtc agttccacca cgggtccgcc agatacagag
     7021 ctagttagct aactagtacc gacgcaggcg cataaaatca gtcatagaca ctagacaatc
     7081 ggacagacac agataagttg ctggccagct tgcctcccgg tggtgggtcg gtggtccctg
     7141 ggcaggggtc tcccgatccc ggacgagccc ccaaatgaaa gacccccgct gacgggtagt
     7201 caatcactca gaggagaccc tcccaaggaa cagcgagacc acaagtcgga tgcaactgca
     7261 agagggttta ttggatacac gggtacccgg gcgactcagt caatcggagg actggcgccc
     7321 cgagtgaggg gttgtgggct cttttattga gctcggggag cagaagcgcg cgaacagaag
     7381 cgagaagcga actgattggt tagttcaaat aaggcacagg gtcatttcag gtccttgggg
     7441 caccctggaa acatctgatg gttctctaga aactgctgag ggctggaccg catctgggga
     7501 ccatctgttc ttggccctga gccggggcag gaactgctta ccacagatat cctgtttggc
     7561 ccatattcag ctgttccatc tgttcttggc cctgagccgg ggcaggaact gcttaccaca
     7621 gatatcctgt ttggcccata ttcaggctgc aggtggcact tttcggggaa atgtgcgcgg
     7681 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     7741 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     7801 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     7861 gctggtgaaa gtaaaagatg ctgaagatca gttg
//
