LOCUS       sequence-323213-        3598 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of three sgRNAs (sgTS5, sgTS6, sgTS7) for targeting to 
            CRISPR-Tag_V2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3598)
  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 3598)
  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: BC1441-mouse U6-3xsgRNAs (TS5, TS6, TS7)
FEATURES             Location/Qualifiers
     source          1..3598
                     /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)."
     protein_bind    complement(976..1009)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(1498..1531)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(1843..2431)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1923..1942)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2602..3393,3394..3462))
                     /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     3225..3244
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3463..3567)
                     /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 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 gttggagtag tcagaataag cgggtttaag
      601 agctatgctg gaaacagcat agcaagttta aataaggcta gtccgttatc aacttgaaaa
      661 agtggcaccg agtcggtgct ttttttctcg agtactagga tccattaggc ggccgcgtgg
      721 ataaccgtat tacgggctaa ttcactccca acgagcaagt tagtaccggg cccgctctag
      781 agatccgacg cgccatctct aggcccgcgc cggccccctc gcacggactt gtgggagaag
      841 ctcggctact cccctgcccc ggttaatttg catataatat ttcctagtaa ctatagaggc
      901 ttaatgtgcg ataaaagaca gataatctgt tctttttaat actagctaca ttttacatga
      961 taggcttgga tttctataac ttcgtatagc atacattata cgaagttata aacagcacaa
     1021 aaggaaactc accctaactg taaagtaatt gtgtgttttg agactataag tatcccttgg
     1081 agaaccacct tgttggggca ccataccgag tgatggttta agagctatgc tggaaacagc
     1141 atagcaagtt taaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
     1201 ctttttttct cgagtactag gatccattag gcggccgcgt ggataaccgt attacccgct
     1261 tgttacaccc tgtgagtgtc gttagtaccg ggcccgctct agagatccga cgcgccatct
     1321 ctaggcccgc gccggccccc tcgcacggac ttgtgggaga agctcggcta ctcccctgcc
     1381 ccggttaatt tgcatataat atttcctagt aactatagag gcttaatgtg cgataaaaga
     1441 cagataatct gttcttttta atactagcta cattttacat gataggcttg gatttctata
     1501 acttcgtata gcatacatta tacgaagtta taaacagcac aaaaggaaac tcaccctaac
     1561 tgtaaagtaa ttgtgtgttt tgagactata agtatccctt ggagaaccac cttgttggct
     1621 accataggca ccacgaggtt taagagctat gctggaaaca gcatagcaag tttaaataag
     1681 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt ctcgagtact
     1741 aggatccatt aggcggccgc gtggataacc gtattacact cacatgtgag caaaaggcca
     1801 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     1861 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1921 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1981 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2041 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2101 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2161 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2221 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2281 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2341 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2401 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2461 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2521 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2581 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2641 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     2701 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagaaccac gctcaccggc
     2761 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     2821 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     2881 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     2941 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3001 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3061 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3121 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3181 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3241 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3301 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3361 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3421 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3481 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3541 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
