LOCUS       sequence-199204-        4767 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor for endogenous N-ter tagging of hLMNA with mClover. Cas9 
            Homologous Recombination Reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4767)
  AUTHORS   Arnoult N, Correia A, Ma J, Merlo A, Garcia-Gomez S, Maric M, 
            Tognetti M, Benner CW, Boulton SJ, Saghatelian A, Karlseder J
  TITLE     Regulation of DNA repair pathway choice in S and G2 phases by the 
            NHEJ inhibitor CYREN.
  JOURNAL   Nature. 2017 Sep 20;549(7673):548-552. doi: 10.1038/nature24023.
  PUBMED    28959974
REFERENCE   2  (bases 1 to 4767)
  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"; journalName: 
            "Nature"; date: "2017-09-20- 20"; volume: "549"; issue: "7673"; 
            pages: "548-552"
COMMENT     Sequence Label: pCAGGS Donor mClover-LMNA
FEATURES             Location/Qualifiers
     source          1..4767
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2273..2289)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2273..2289)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2286..2308)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2297..2313
                     /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(2321..2327,2328..2345,2346..2351))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2366..2387
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2446..2641
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2492..2627
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2647..2781
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2684..2703)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2738..2757
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2849..2866)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3020..3608)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3100..3119)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3779..4570,4571..4639))
                     /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     4402..4421
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4640..4744)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctggtcgaca ttgattagtc gactctcagt accactttcc tggctcccag ccctgcctac
       61 ctgaccctct cccttgcttt gcgcccactt ccctctcttt ctccccgacc cttttgccca
      121 cccactctcc ctccttggct ctgccctcta gcccagaagg tctgaggcaa tgggggcaag
      181 cttggagccg acagtgctga gcaggcagga gccaagagag gggaagcttg agcctcacgc
      241 agttaggggt gcgctggaga gggtggggcc cgactccgcc acaccccaac ggtccttccc
      301 cctcctcacc actcccgccc ccacccccaa tggatctggg actgcccctt taagagtagt
      361 ggcccctcct cccttcagag gaggacctat tagagccttt gccccggcgt cggtgactca
      421 gtgttcgcgg gagcgccgca cctacaccag ccaacccaga tcccgaggtc cgacagcgcc
      481 cggcccagat ccccacgcct gccaggagca agccgagagc cagccggccg gcgcactccg
      541 actccgagca gtctctgtcc ttcgacccga gccccgcgcc ctttccggga cccctgcccc
      601 gcgggcagcg ctgccaacct gccggccatg gaggtcagta aaggtgagga gcttttcact
      661 ggggtagtgc cgattctcgt ggagctggac ggtgatgtaa acggacataa atttagtgtt
      721 cgcggggaag gcgaaggaga tgcaacaaat ggcaagctta cacttaaatt catttgcacc
      781 acgggcaagt tgccagtgcc ttggcctaca ttggttacga cgtttggtta cggtgtggcg
      841 tgtttcagtc ggtatccaga tcacatgaaa caacacgatt tttttaaatc agccatgccg
      901 gaaggttacg tgcaagagcg gacgataagt tttaaggacg acgggacata caagactagg
      961 gctgaggtca aatttgaggg cgacaccctg gtgaatcgga ttgagcttaa ggggatagat
     1021 tttaaagaag acggcaacat cctcggacac aagctggagt acaactttaa ttcacataat
     1081 gtgtacataa ctgcagataa acaaaagaat ggaataaaag ccaactttaa gatccggcac
     1141 aacgtagaag atggtagcgt ccagctggcc gaccactacc agcagaacac gcccattgga
     1201 gatggacccg ttcttttgcc tgacaaccat tacctcagcc accaatccgc actttcaaaa
     1261 gacccaaatg aaaaaagaga tcatatggta ctgctggagt tcgtcactgc agctggtatt
     1321 actcacggca tggatgaact gtataagacc ccgtcccagc ggcgcgccac ccgcagcggg
     1381 gcgcaggcca gctccactcc gctgtcgccc acccgcatca cccggctgca ggagaaggag
     1441 gacctgcagg agctcaatga tcgcttggcg gtctacatcg accgtgtgcg ctcgctggaa
     1501 acggagaacg cagggctgcg ccttcgcatc accgagtctg aagaggtggt cagccgcgag
     1561 gtgtccggca tcaaggccgc ctacgaggcc gagctcgggg atgcccgcaa gacccttgac
     1621 tcagtagcca aggagcgcgc ccgcctgcag ctggagctga gcaaagtgcg tgaggagttt
     1681 aaggagctga aagcgcggtg agttcgccca ggtggctgcg tgcctggcgg ggagtggaga
     1741 gggcggcggg ccggcgcccc tggccggccg caggaaggga gtgagagggc ctggaggccg
     1801 ataactttgc catagtctcc tccctccccg gaactgcccc cagcgggtga ctggcagtgt
     1861 caaggggaat tgtcaagaca ggacagagag ggaagtggtg gtctctggga gagggtcggg
     1921 gaggatataa ggaatggtgg gggtatcagg gacagcgact cataatcagc cataccacag
     1981 caacatatgc catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta
     2041 tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta
     2101 gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt
     2161 acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc
     2221 tcttctctta tggagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc
     2281 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     2341 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     2401 cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc
     2461 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     2521 ccattctccg ccccatggct gactaatttt ttttatttat acagaggccg aggccgcctc
     2581 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     2641 aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     2701 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     2761 gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2821 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2881 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2941 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3001 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3061 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3121 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3181 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3241 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3301 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3361 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3421 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3481 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3541 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3601 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3661 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3721 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3781 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3841 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     3901 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     3961 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4021 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4081 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4141 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4201 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     4261 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     4321 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4381 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4441 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     4501 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4561 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4621 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     4681 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     4741 cgcgcacatt tccccgaaaa gtgccac
//
