LOCUS       sequence-392213-        4785 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to insert [mTagBFP-stop-12XMS2] into the 
            C-terminus of human HSPB1 gene; 12XMS2 locates in the 3' UTR region.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4785)
  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 4785)
  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: pCA20-HSPB1 5'HA-mTagBFP-3'-UTR 12XMS2V5-3'HA ...
FEATURES             Location/Qualifiers
     source          1..4785
                     /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"
     CDS             750..1451
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     primer_bind     2796..2815
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     rep_origin      complement(3030..3618)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3110..3129)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3789..4580,4581..4649))
                     /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     4412..4431
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4650..4754)
                     /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 aggacggctg cctcatctac aacgtcaaga tcagaggggt gaacttcaca tccaacggcc
     1141 ctgtgatgca gaagaaaaca ctcggctggg aggccttcac cgagacgctg taccccgctg
     1201 acggcggcct ggaaggcaga aacgacatgg ccctgaagct cgtgggcggg agccatctga
     1261 tcgcaaacat caagaccaca tatagatcca agaaacccgc taagaacctc aagatgcctg
     1321 gcgtctacta tgtggactac agactggaaa gaatcaagga ggccaacaac gagacctacg
     1381 tcgagcagca cgaggtggca gtggccagat actgcgacct ccctagcaaa ctggggcaca
     1441 agcttaatta agcggccgcc cggtaaccta caaacgggtg gaggatcacc ccacccgaca
     1501 cttcacaatc aaggggtaca atacacaagg gtggaggaac accccaccct ccagacacat
     1561 tacacagaaa tccaatcaaa cagaagcacc atcagggctt ctgctaccaa atttatctca
     1621 aaaaactaca acaaggaatc accatcaggg attccctgtg caatatacgt caaacgaggg
     1681 ccacgacggg aggacgatca cgcctcccga atatcggcat gtctggcttt cgaattcagt
     1741 gcgtggagca tcagcccacg cagccaatca gagtcgaata caagtcgact ttcgcgaaga
     1801 gcatcagcct tcgcgccatt cttacacaaa ccacactctc ccctacagga acagcatcag
     1861 cgttcctgcc cagtacccaa ctcaagaaaa tttatgtccc catgcagcat cagcgcatgg
     1921 gccccaagaa tacatcccca acaaaatcac atccgagcac caacagggct cggagtgttg
     1981 tttcttgtcc aactggacaa accctccatg gaccatcagg ccatggactc tcaccaacaa
     2041 gacaaaaact actcttctcg aagcagcatc agcgcttcga aacactcgag catacattgt
     2101 gcctatttct tgggtggacg gaaccggtgc tggatccggt tagagcctta gcccggatgc
     2161 ccacccctgc tgccgccact ggctgtgcct cccccgccac ctgtgtgttc ttttgataca
     2221 tttatcttct gtttttctca aataaagttc aaagcaacca cctgtcactg gcccaggccc
     2281 tggtgtttgt ggaaggaagc ctcaggcacc tgccatttgc tggctttcag gagtcatctt
     2341 tgctcaggcc cgtgctgggc catgtgggta cactggtgta ggttgctgga cacaggctga
     2401 ctcacatcca taaagacaga ggtcttaggg ccgggcgcag tggctcatac ctacaatccc
     2461 agcactttgg ggggttgaag caggaggagt gcttgaagcc aagagttcta gaccagcctg
     2521 gacaacatag taagactgtc tctaaaaaat aaaaattagg cagggtggta ctgcacgcct
     2581 gtagtcccag ctactcagga ggctgaggca ggaggatcgc ttgagcccag gagttgaagg
     2641 gtacagtgag ctaacatcgt gccgatatcg gagcttactg agactcttcg ggcccgggcg
     2701 acggatggtg atccccctgg ccagtgcacg tctgctgtca gataaagtct cccgtgaact
     2761 ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag
     2821 tgtgccggtc tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat
     2881 caaaaacgcc attaacctga tgttctgggg aatataaatg tcaggctccc ttatacacag
     2941 ccagtctgca ggtcgacggt ctcgactcac atgtgagcaa aaggccagca aaaggccagg
     3001 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3061 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3121 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3181 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3241 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3301 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3361 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3421 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     3481 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     3541 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     3601 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     3661 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     3721 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     3781 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3841 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3901 tctggcccca gtgctgcaat gataccgcga gaaccacgct caccggctcc agatttatca
     3961 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4021 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4081 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4141 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4201 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4261 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4321 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4381 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4441 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4501 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4561 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4621 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     4681 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4741 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
