LOCUS       sequence-392219-        4878 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to insert TriTag (mTagBFP harbors 12XMS2 in its
            intron) into the C-terminus of human HSPB1 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4878)
  AUTHORS   Liang Y, Xu H, Cheng T, Fu Y, Huang H, Qian W, Wang J, Zhou Y, Qian 
            P, Yin Y, Xu P, Zou W, Chen B
  TITLE     Gene activation guided by nascent RNA-bound transcription factors.
  JOURNAL   Nat Commun. 2022 Nov 28;13(1):7329. doi: 10.1038/s41467-022-35041-7.
  PUBMED    36443367
REFERENCE   2  (bases 1 to 4878)
  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: "2022-11-28"; volume: "13(1):7329. doi"; pages: " 
            10.1038/s41467-022-35041-7"
COMMENT     Sequence Label: pCA19-HSPB1 5'HA-TriTag (mTagBFP)-3'HA (HDR do...
FEATURES             Location/Qualifiers
     source          1..4878
                     /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     2889..2908
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     rep_origin      complement(3123..3711)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3203..3222)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3882..4673,4674..4742))
                     /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     4505..4524
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4743..4847)
                     /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 cgtggagctt
      241 actgagactc ttcggggaat tctcctggct gcttccaagc aggaggtggg gcctctggcc
      301 tagcggggcc gaaaggcagt cccctccccc gcagtctgat ttccctcttc cccccaaagg
      361 caagcacgag gagcggcagg acgagcatgg ctacatctcc cggtgcttca cgcggaaata
      421 cacgtgagtc ctggcgccag gtcggggtgg gtgggtggcg tgggggtggg gtcagggaag
      481 agggcacagg gacccacccg gtgtgtaatg taacgcttgc ctttcctctc tgcacgtcca
      541 ggctgccccc cggtgtggac cccacccaag tttcctcctc cctgtcccct gagggcacac
      601 tgaccgtgga ggcccccatg cccaagctag ccacgcagtc caacgagatc accatcccag
      661 tcaccttcga gtcgcgggcc cagcttgggg gcccagaagc tgcaaaatcc gatgagactg
      721 ccgccaaggg atcgggtggt accagtggca tgagcgagct gattaaggag aacatgcaca
      781 tgaagctgta catggagggc accgtggaca accatcactt caagtgcaca tccgagggcg
      841 aaggcaagcc ctacgagggc acccagacca tgagaatcaa ggtggtcgag ggcggccctc
      901 tccccttcgc cttcgacatc ctggctacta gcttcctcta cggcagcaag accttcatca
      961 accacaccca gggcatcccc gacttcttca agcagtcctt ccctgagggc ttcacatggg
     1021 agagagtcac cacatacgaa gacgggggcg tgctgaccgc tacccaggac accagcctcc
     1081 aggacgtaag tatgaaattc agggatacgg ccaccttgtt ggccggtaac ctacaaacgg
     1141 gtggaggatc accccacccg acacgcaccg atgctctccg aggaggcaca agggtggagg
     1201 aacaccccac cctccaggca ccgatgctct ccgaggaggc aaacagaagc accatcaggg
     1261 cttctgctac gcaccgatgc tctccgagga ggaacaagga atcaccatca gggattccct
     1321 gtggcaccga tgctctccga ggaggacgac gggaggacga tcacgcctcc cgaatagcac
     1381 cgatgctctc cgaggaggtc agtgcgtgga gcatcagccc acgcagccag caccgatgct
     1441 ctccgaggag gctttcgcga agagcatcag ccttcgcgcc atgcaccgat gctctccgag
     1501 gaggcctaca ggaacagcat cagcgttcct gcccagcacc gatgctctcc gaggaggtgt
     1561 ccccatgcag catcagcgca tgggccccgc accgatgctc tccgaggagg cacatccgag
     1621 caccaacagg gctcggagtg tgcaccgatg ctctccgagg aggaccctcc atggaccatc
     1681 aggccatgga ctctgcaccg atgctctccg aggaggcttc tcgaagcagc atcagcgctt
     1741 cgaaacagca ccgatgctct ccgaggaggt tctccacctt gttggcatat ttgccaaata
     1801 gtggaaatgt gaagtactga caaaactttt ccctttttca atctaatagg gctgcctcat
     1861 ctacaacgtc aagatcagag gggtgaactt cacatccaac ggccctgtga tgcagaagaa
     1921 aacactcggc tgggaggcct tcaccgagac gctgtacccc gctgacggcg gcctggaagg
     1981 cagaaacgac atggccctga agctcgtggg cgggagccat ctgatcgcaa acatcaagac
     2041 cacatataga tccaagaaac ccgctaagaa cctcaagatg cctggcgtct actatgtgga
     2101 ctacagactg gaaagaatca aggaggccaa caacgagacc tacgtcgagc agcacgaggt
     2161 ggcagtggcc agatactgcg acctccctag caaactgggg cacaagctta atggaaccgg
     2221 tgctggatcc ggttagagcc ttagcccgga tgcccacccc tgctgccgcc actggctgtg
     2281 cctcccccgc cacctgtgtg ttcttttgat acatttatct tctgtttttc tcaaataaag
     2341 ttcaaagcaa ccacctgtca ctggcccagg ccctggtgtt tgtggaagga agcctcaggc
     2401 acctgccatt tgctggcttt caggagtcat ctttgctcag gcccgtgctg ggccatgtgg
     2461 gtacactggt gtaggttgct ggacacaggc tgactcacat ccataaagac agaggtctta
     2521 gggccgggcg cagtggctca tacctacaat cccagcactt tggggggttg aagcaggagg
     2581 agtgcttgaa gccaagagtt ctagaccagc ctggacaaca tagtaagact gtctctaaaa
     2641 aataaaaatt aggcagggtg gtactgcacg cctgtagtcc cagctactca ggaggctgag
     2701 gcaggaggat cgcttgagcc caggagttga agggtacagt gagctaacat cgtgccgata
     2761 tcggagctta ctgagactct tcgggcccgg gcgacggatg gtgatccccc tggccagtgc
     2821 acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata tcggggatga
     2881 aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtctccgtta tcggggaaga
     2941 agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg
     3001 gggaatataa atgtcaggct cccttataca cagccagtct gcaggtcgac ggtctcgact
     3061 cacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3121 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3181 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3241 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3301 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3361 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3421 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3481 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3541 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3601 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3661 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3721 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3781 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3841 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     3901 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3961 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4021 cgagaaccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4081 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4141 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4201 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4261 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4321 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4381 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4441 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     4501 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4561 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4621 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4681 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     4741 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4801 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     4861 aaagtgccac ctgacgtc
//
