LOCUS       sequence-392209-        5119 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to insert NarTag (mTagBFP harbors 12XPP7 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 5119)
  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 5119)
  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: pCA21-HSPB1 5'HA-NarTag (12XPP7)-3'HA (HDR donor)
FEATURES             Location/Qualifiers
     source          1..5119
                     /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     3130..3149
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     rep_origin      complement(3364..3952)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3444..3463)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4123..4914,4915..4983))
                     /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     4746..4765
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4984..5088)
                     /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 ggccagatat acgcgttgac
     1141 gttagtaccg ggcccgctct agtacaaacc agcagagcat atgggctcgc tggacacgca
     1201 ccgatgctct ccgaggaggc acaggagcag acgatatggc gtcgctcccc aggcaccgat
     1261 gctctccgag gaggcaaacc agcagagcat atgggctcgc tggctacgca ccgatgctct
     1321 ccgaggagga acaggagcag acgatatggc gtcgctcctg tggcaccgat gctctccgag
     1381 gaggacgagc cgcgtggata accgtattac gggctaattc actcccaacg agcaagttag
     1441 taccgggccc gctctagtac aaaccagcag agcatatggg ctcgctggac acgcaccgat
     1501 gctctccgag gaggcacagg agcagacgat atggcgtcgc tccccaggca ccgatgctct
     1561 ccgaggaggc aaaccagcag agcatatggg ctcgctggct acgcaccgat gctctccgag
     1621 gaggaacagg agcagacgat atggcgtcgc tcctgtggca ccgatgctct ccgaggagga
     1681 cgagccgcgt ggataaccgt attacccgct tgttacaccc tgtgagtgtc gttagtaccg
     1741 ggcccgctct agtacaaacc agcagagcat atgggctcgc tggacacgca ccgatgctct
     1801 ccgaggaggc acaggagcag acgatatggc gtcgctcccc aggcaccgat gctctccgag
     1861 gaggcaaacc agcagagcat atgggctcgc tggctacgca ccgatgctct ccgaggagga
     1921 acaggagcag acgatatggc gtcgctcctg tggcaccgat gctctccgag gaggacgagc
     1981 cgcgtggata accgtattac actcacatgt gagcccacct tgttggcata tttgccaaat
     2041 agtggaaatg tgaagtactg acaaaacttt tccctttttc aatctaatag ggctgcctca
     2101 tctacaacgt caagatcaga ggggtgaact tcacatccaa cggccctgtg atgcagaaga
     2161 aaacactcgg ctgggaggcc ttcaccgaga cgctgtaccc cgctgacggc ggcctggaag
     2221 gcagaaacga catggccctg aagctcgtgg gcgggagcca tctgatcgca aacatcaaga
     2281 ccacatatag atccaagaaa cccgctaaga acctcaagat gcctggcgtc tactatgtgg
     2341 actacagact ggaaagaatc aaggaggcca acaacgagac ctacgtcgag cagcacgagg
     2401 tggcagtggc cagatactgc gacctcccta gcaaactggg gcacaagctt aatggaaccg
     2461 gtgctggatc cggttagagc cttagcccgg atgcccaccc ctgctgccgc cactggctgt
     2521 gcctcccccg ccacctgtgt gttcttttga tacatttatc ttctgttttt ctcaaataaa
     2581 gttcaaagca accacctgtc actggcccag gccctggtgt ttgtggaagg aagcctcagg
     2641 cacctgccat ttgctggctt tcaggagtca tctttgctca ggcccgtgct gggccatgtg
     2701 ggtacactgg tgtaggttgc tggacacagg ctgactcaca tccataaaga cagaggtctt
     2761 agggccgggc gcagtggctc atacctacaa tcccagcact ttggggggtt gaagcaggag
     2821 gagtgcttga agccaagagt tctagaccag cctggacaac atagtaagac tgtctctaaa
     2881 aaataaaaat taggcagggt ggtactgcac gcctgtagtc ccagctactc aggaggctga
     2941 ggcaggagga tcgcttgagc ccaggagttg aagggtacag tgagctaaca tcgtgccgat
     3001 atcggagctt actgagactc ttcgggcccg ggcgacggat ggtgatcccc ctggccagtg
     3061 cacgtctgct gtcagataaa gtctcccgtg aactttaccc ggtggtgcat atcggggatg
     3121 aaagctggcg catgatgacc accgatatgg ccagtgtgcc ggtctccgtt atcggggaag
     3181 aagtggctga tctcagccac cgcgaaaatg acatcaaaaa cgccattaac ctgatgttct
     3241 ggggaatata aatgtcaggc tcccttatac acagccagtc tgcaggtcga cggtctcgac
     3301 tcacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3361 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3421 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3481 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3541 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3601 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3661 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3721 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3781 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3841 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3901 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3961 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4021 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4081 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4141 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4201 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4261 gcgagaacca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4321 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4381 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4441 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4501 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4561 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4621 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4681 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4741 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4801 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4861 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4921 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4981 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5041 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5101 aaaagtgcca cctgacgtc
//
