LOCUS       sequence-323220-        4922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to insert mCherry-harboring CRISPR-TagV2 into 
            the N-terminus of human LMNA gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4922)
  AUTHORS   Chen B, Zou W, Xu H, Liang Y, Huang B
  TITLE     Efficient labeling and imaging of protein-coding genes in living 
            cells using CRISPR-Tag.
  JOURNAL   Nat Commun. 2018 Nov 29;9(1):5065. doi: 10.1038/s41467-018-07498-y.
  PUBMED    30498221
REFERENCE   2  (bases 1 to 4922)
  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: "2018-11-29"; volume: "9(1):5065. doi"; pages: " 
            10.1038/s41467-018-07498-y"
COMMENT     Sequence Label: BC1455-LMNA 5'HA-mCherry-CRISPR-Tag_V2-3'HA (H...
FEATURES             Location/Qualifiers
     source          1..4922
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(942..960)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1210..1229
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2933..2952
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     rep_origin      complement(3167..3755)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3247..3266)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3926..4717,4718..4786))
                     /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     4549..4568
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4787..4891)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattccatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacaga
      241 gaccgcggcc gcccgctggg acggggtctc catggcccct ccttggctct gccctctagc
      301 ccagaaggtc tgaggcaatg ggggcaagct tggagccgac agtgctgagc aggcaggagc
      361 caagagaggg gaagcttgag cctcacgcag ttaggggtgc gctggagagg gtggggcccg
      421 actccgccac accccaacgg tccttccccc tcctcaccac tcccgccccc acccccaatg
      481 gatctgggac tgccccttta agagtagtgg cccctcctcc cttcagagga ggacctatta
      541 gagcctttgc cctggcgtcg gtgactcagt gttcgcggga gcgccgcacc tacaccagcc
      601 aacccagatc ccgaggtccg acagcgcccg gcccagatcc ccacgcctgc caggagcaag
      661 ccgagagcca gccggccggc gcactccgac tccgagcagt ctctgtcctt cgacccgagc
      721 cccgcgccct ttccgggacc cctgccccgc gggcagcgct gccaacctgc cggccggatc
      781 gggtgggact agtggcatgg tgagcaaggg cgaggaggat aacatggcca tcatcaagga
      841 gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt tcgagatcga
      901 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
      961 gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt acggctccaa
     1021 ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct tccccgaggg
     1081 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg tgacccagga
     1141 ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc
     1201 ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct ccgagcggat
     1261 gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc tgaaggacgg
     1321 cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc
     1381 cggcgccgta agtatgaaat tcagggatac ggccaccttg ttggcgggcc agatatacgc
     1441 gttgacgagt agtcagaata agcggaggcc ctgaggggca ccataccgag tgatggggcc
     1501 catcagctac cataggcacc acgagcggca ggggctaatt cactcccaac gagcaagagt
     1561 agtcagaata agcggaggcc ctgaggggca ccataccgag tgatggggcc catcagctac
     1621 cataggcacc acgagcggca gccttgatct gtggatctac cagaacgagt agtcagaata
     1681 agcggaggcc ctgaggggca ccataccgag tgatggggcc catcagctac cataggcacc
     1741 acgagcggca gggcagaact acacaccagg gccagtgagt agtcagaata agcggaggcc
     1801 ctgaggggca ccataccgag tgatggggcc catcagctac cataggcacc acgagcggca
     1861 gcatcggcat ctttgacgca agacacgagt agtcagaata agcggaggcc ctgaggggca
     1921 ccataccgag tgatggggcc catcagctac cataggcacc acgagcggca gccgcttgtt
     1981 acaccctgtg agtgtcgagt agtcagaata agcggaggcc ctgaggggca ccataccgag
     2041 tgatggggcc catcagctac cataggcacc acgagcggca ggcagtagta tctttgcaag
     2101 tgactcaccc accttgttgg catatttgcc aaatagtgga aatgtgaagt actgacaaaa
     2161 cttttccctt tttcaatcta atagtacaac gtcaacatca agttggacat cacctcccac
     2221 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
     2281 ggcatggacg agctgtacaa gggaaccggt gctggaagtg gtgagacccc gtcccaacga
     2341 cgcgccaccc gcagcggggc gcaggccagc tccactccgc tgtcgcccac ccgcatcacc
     2401 cggctgcagg agaaggagga cctgcaggag ctcaatgatc gcttggcggt ctacatcgac
     2461 cgtgtgcgct cgctggaaac ggagaacgca gggctgcgcc ttcgcatcac cgagtctgaa
     2521 gaggtggtca gccgcgaggt gtccggcatc aaggccgcct acgaggccga gctcggggat
     2581 gcccgcaaga cccttgactc agtagccaag gagcgcgccc gcctgcagct ggagctgagc
     2641 aaagtgcgtg aggagtttaa ggagctgaaa gcgcggtgag ttcgcccagg tggctgcgtg
     2701 cctggcgggg agtggagagg gcggcgggcc ggcgcccctg gccggccgca ggaagggagt
     2761 gagagggcct ggaggccgat aactttgcca tagtctcctc cctccgccat ggagaccccg
     2821 tcccagcggc ccgggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat
     2881 aaagtctccc gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg
     2941 accaccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc
     3001 caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat ataaatgtca
     3061 ggctccctta tacacagcca gtctgcaggt cgacggtctc gactcacatg tgagcaaaag
     3121 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3181 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3241 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3301 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3361 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3421 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3481 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3541 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3601 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3661 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3721 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3781 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3841 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3901 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3961 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4021 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagaa ccacgctcac
     4081 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4141 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4201 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4261 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4321 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4381 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4441 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4501 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4561 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4621 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4681 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4741 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4801 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4861 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     4921 tc
//
