LOCUS       sequence-323255-        2576 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  U6-driven sgRNA targeting H2B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2576)
  AUTHORS   Chen B, Zou W, Xu H, Liang Y, Huang B
  TITLE     Efficient labeling and imaging of protein-coding genes in living 
            cells using CRISPR-Tag.
  JOURNAL   Nat Commun. 2018 Nov 29;9(1):5065. doi: 10.1038/s41467-018-07498-y.
  PUBMED    30498221
REFERENCE   2  (bases 1 to 2576)
  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: "2018-11-29"; volume: "9(1):5065. doi"; pages: " 
            10.1038/s41467-018-07498-y"
COMMENT     Sequence Label: BC1364-mouse U6-sgH2B
FEATURES             Location/Qualifiers
     source          1..2576
                     /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"
     protein_bind    complement(455..488)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     492..512
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     rep_origin      complement(821..1409)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(901..920)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1580..2371,2372..2440))
                     /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     2203..2222
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2441..2545)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctacc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattccatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacgtt
      241 agtaccgggc ccgctctaga gatccgacgc gccatctcta ggcccgcgcc ggccccctcg
      301 cacggacttg tgggagaagc tcggctactc ccctgccccg gttaatttgc atataatatt
      361 tcctagtaac tatagaggct taatgtgcga taaaagacag ataatctgtt ctttttaata
      421 ctagctacat tttacatgat aggcttggat ttctataact tcgtatagca tacattatac
      481 gaagttataa acagcacaaa aggaaactca ccctaactgt aaagtaattg tgtgttttga
      541 gactataagt atcccttgga gaaccacctt gttggcgagc gccaggtccc ggcagtttaa
      601 gagctatgct ggaaacagca tagcaagttt aaataaggct agtccgttat caacttgaaa
      661 aagtggcacc gagtcggtgc tttttttctc gagtactagg atccattagg cggccgcgtg
      721 gataaccgta ttacgggcta attcactccc aacgaactca catgtgagca aaaggccagc
      781 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
      841 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
      901 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
      961 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     1021 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     1081 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     1141 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     1201 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     1261 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     1321 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     1381 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     1441 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     1501 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     1561 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     1621 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     1681 agggcttacc atctggcccc agtgctgcaa tgataccgcg agaaccacgc tcaccggctc
     1741 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     1801 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     1861 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     1921 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     1981 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     2041 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     2101 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     2161 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     2221 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     2281 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     2341 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     2401 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     2461 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     2521 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
