LOCUS       sequence-323203-        4889 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to insert TriTag (mCherry) into the N-terminus 
            of human LMNA gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4889)
  AUTHORS   Xu H, Wang J, Liang Y, Fu Y, Li S, Huang J, Xu H, Zou W, Chen B
  TITLE     TriTag: an integrative tool to correlate chromatin dynamics and gene
            expression in living cells.
  JOURNAL   Nucleic Acids Res. 2020 Oct 26. pii: 5940501. doi: 
            10.1093/nar/gkaa906.
  PUBMED    33104788
REFERENCE   2  (bases 1 to 4889)
  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: "Nucleic 
            Acids Res. 2020 Oct 26. pii: 5940501. doi: 10.1093/nar/gkaa906."
COMMENT     Sequence Label: PW124-LMNA 5'HA-TriTag (mCherry)-3'HA (HDR donor)
FEATURES             Location/Qualifiers
     source          1..4889
                     /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     2900..2919
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     rep_origin      complement(3134..3722)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3214..3233)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3893..4684,4685..4753))
                     /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     4516..4535
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4754..4858)
                     /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 cccggcgtcg 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 ttggccggta acctacaaac
     1441 gggtggagga tcaccccacc cgacacgcac cgatgctctc cgaggaggca caagggtgga
     1501 ggaacacccc accctccagg caccgatgct ctccgaggag gcaaacagaa gcaccatcag
     1561 ggcttctgct acgcaccgat gctctccgag gaggaacaag gaatcaccat cagggattcc
     1621 ctgtggcacc gatgctctcc gaggaggacg acgggaggac gatcacgcct cccgaatagc
     1681 accgatgctc tccgaggagg tcagtgcgtg gagcatcagc ccacgcagcc agcaccgatg
     1741 ctctccgagg aggctttcgc gaagagcatc agccttcgcg ccatgcaccg atgctctccg
     1801 aggaggccta caggaacagc atcagcgttc ctgcccagca ccgatgctct ccgaggaggt
     1861 gtccccatgc agcatcagcg catgggcccc gcaccgatgc tctccgagga ggcacatccg
     1921 agcaccaaca gggctcggag tgtgcaccga tgctctccga ggaggaccct ccatggacca
     1981 tcaggccatg gactctgcac cgatgctctc cgaggaggct tctcgaagca gcatcagcgc
     2041 ttcgaaacag caccgatgct ctccgaggag gttctccacc ttgttggcat atttgccaaa
     2101 tagtggaaat gtgaagtact gacaaaactt ttcccttttt caatctaata gtacaacgtc
     2161 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
     2221 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaaggg aaccggtgct
     2281 ggaagtggtg agaccccgtc ccaacgacgc gccacccgca gcggggcgca ggccagctcc
     2341 actccgctgt cgcccacccg catcacccgg ctgcaggaga aggaggacct gcaggagctc
     2401 aatgatcgct tggcggtcta catcgaccgt gtgcgctcgc tggaaacgga gaacgcaggg
     2461 ctgcgccttc gcatcaccga gtctgaagag gtggtcagcc gcgaggtgtc cggcatcaag
     2521 gccgcctacg aggccgagct cggggatgcc cgcaagaccc ttgactcagt agccaaggag
     2581 cgcgcccgcc tgcagctgga gctgagcaaa gtgcgtgagg agtttaagga gctgaaagcg
     2641 cggtgagttc gcccaggtgg ctgcgtgcct ggcggggagt ggagagggcg gcgggccggc
     2701 gcccctggcc ggccgcagga agggagtgag agggcctgga ggccgataac tttgccatag
     2761 tctcctccct ccgccatgga gaccccgtcc cagcggcccg ggcgacggat ggtgatcccc
     2821 ctggccagtg cacgtctgct gtcagataaa gtctcccgtg aactttaccc ggtggtgcat
     2881 atcggggatg aaagctggcg catgatgacc accgatatgg ccagtgtgcc ggtctccgtt
     2941 atcggggaag aagtggctga tctcagccac cgcgaaaatg acatcaaaaa cgccattaac
     3001 ctgatgttct ggggaatata aatgtcaggc tcccttatac acagccagtc tgcaggtcga
     3061 cggtctcgac tcacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3121 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3181 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3241 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3301 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3361 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3421 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3481 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3541 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3601 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3661 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3721 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3781 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3841 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3901 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     3961 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4021 caatgatacc gcgagaacca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4081 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4141 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4201 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4261 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4321 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4381 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4441 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     4501 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     4561 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     4621 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     4681 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4741 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4801 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     4861 catttccccg aaaagtgcca cctgacgtc
//
