LOCUS       sequence-323202-        4880 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to insert TriTag (mTagBFP) into the N-terminus 
            of human LMNA gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4880)
  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 4880)
  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: PHY98-LMNA 5'HA-TriTag (mTagBFP)-3'HA (HDR donor)
FEATURES             Location/Qualifiers
     source          1..4880
                     /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     2891..2910
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     rep_origin      complement(3125..3713)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3205..3224)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3884..4675,4676..4744))
                     /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     4507..4526
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4745..4849)
                     /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 agtggcatga gcgagctgat taaggagaac atgcacatga agctgtacat
      841 ggagggcacc gtggacaacc atcacttcaa gtgcacatcc gagggcgaag gcaagcccta
      901 cgagggcacc cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt
      961 cgacatcctg gctactagct tcctctacgg cagcaagacc ttcatcaacc acacccaggg
     1021 catccccgac ttcttcaagc agtccttccc tgagggcttc acatgggaga gagtcaccac
     1081 atacgaagac gggggcgtgc tgaccgctac ccaggacacc agcctccagg acgtaagtat
     1141 gaaattcagg gatacggcca ccttgttggc cggtaaccta caaacgggtg gaggatcacc
     1201 ccacccgaca cgcaccgatg ctctccgagg aggcacaagg gtggaggaac accccaccct
     1261 ccaggcaccg atgctctccg aggaggcaaa cagaagcacc atcagggctt ctgctacgca
     1321 ccgatgctct ccgaggagga acaaggaatc accatcaggg attccctgtg gcaccgatgc
     1381 tctccgagga ggacgacggg aggacgatca cgcctcccga atagcaccga tgctctccga
     1441 ggaggtcagt gcgtggagca tcagcccacg cagccagcac cgatgctctc cgaggaggct
     1501 ttcgcgaaga gcatcagcct tcgcgccatg caccgatgct ctccgaggag gcctacagga
     1561 acagcatcag cgttcctgcc cagcaccgat gctctccgag gaggtgtccc catgcagcat
     1621 cagcgcatgg gccccgcacc gatgctctcc gaggaggcac atccgagcac caacagggct
     1681 cggagtgtgc accgatgctc tccgaggagg accctccatg gaccatcagg ccatggactc
     1741 tgcaccgatg ctctccgagg aggcttctcg aagcagcatc agcgcttcga aacagcaccg
     1801 atgctctccg aggaggttct ccaccttgtt ggcatatttg ccaaatagtg gaaatgtgaa
     1861 gtactgacaa aacttttccc tttttcaatc taatagggct gcctcatcta caacgtcaag
     1921 atcagagggg tgaacttcac atccaacggc cctgtgatgc agaagaaaac actcggctgg
     1981 gaggccttca ccgagacgct gtaccccgct gacggcggcc tggaaggcag aaacgacatg
     2041 gccctgaagc tcgtgggcgg gagccatctg atcgcaaaca tcaagaccac atatagatcc
     2101 aagaaacccg ctaagaacct caagatgcct ggcgtctact atgtggacta cagactggaa
     2161 agaatcaagg aggccaacaa cgagacctac gtcgagcagc acgaggtggc agtggccaga
     2221 tactgcgacc tccctagcaa actggggcac aagcttaatg gaaccggtgc tggaagtggt
     2281 gagaccccgt cccaacgacg cgccacccgc agcggggcgc aggccagctc cactccgctg
     2341 tcgcccaccc gcatcacccg gctgcaggag aaggaggacc tgcaggagct caatgatcgc
     2401 ttggcggtct acatcgaccg tgtgcgctcg ctggaaacgg agaacgcagg gctgcgcctt
     2461 cgcatcaccg agtctgaaga ggtggtcagc cgcgaggtgt ccggcatcaa ggccgcctac
     2521 gaggccgagc tcggggatgc ccgcaagacc cttgactcag tagccaagga gcgcgcccgc
     2581 ctgcagctgg agctgagcaa agtgcgtgag gagtttaagg agctgaaagc gcggtgagtt
     2641 cgcccaggtg gctgcgtgcc tggcggggag tggagagggc ggcgggccgg cgcccctggc
     2701 cggccgcagg aagggagtga gagggcctgg aggccgataa ctttgccata gtctcctccc
     2761 tccgccatgg agaccccgtc ccagcggccc gggcgacgga tggtgatccc cctggccagt
     2821 gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat
     2881 gaaagctggc gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa
     2941 gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc
     3001 tggggaatat aaatgtcagg ctcccttata cacagccagt ctgcaggtcg acggtctcga
     3061 ctcacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3121 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3181 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3241 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3301 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3361 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3421 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3481 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3541 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3601 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3661 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3721 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3781 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3841 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3901 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3961 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4021 cgcgagaacc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4081 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4141 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4201 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4261 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4321 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4381 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4441 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     4501 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     4561 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4621 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4681 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4741 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4801 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     4861 gaaaagtgcc acctgacgtc
//
