LOCUS       sequence-323199-        4748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR donor plasmid to insert mCherry-CRISPR-TagV2 into the 
            C-terminus of human H2B gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4748)
  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 4748)
  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: BC1450-H2B 5'HA-mCherry-CRISPR-Tag_V2-3'HA (HD...
FEATURES             Location/Qualifiers
     source          1..4748
                     /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     complement(845..863)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1113..1132
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2759..2778
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     rep_origin      complement(2993..3581)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3073..3092)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3752..4543,4544..4612))
                     /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     4375..4394
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4613..4717)
                     /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 cgttgacaga
      241 gaccgcggcc gcccctgccg ggacctggcg ctcgcctaag ccgcgtttgt actgtgtctt
      301 accatgcctg aaccggcaaa atccgctccg gcccctaaaa agggctccaa gaaagccgtc
      361 accaaagccc agaagaaaga cggcaagaag cgcaagcgca gccgcaaaga gagctactcc
      421 atctacgtgt acaaggtgct gaagcaggtc caccccgaca ccggcatctc gtccaaggcc
      481 atgggcatca tgaactcctt cgtcaacgac atcttcgagc gcatcgcggg agaggcttcc
      541 cgcctggcgc actacaacaa gcgctccacc atcacatccc gcgagatcca gacggccgtg
      601 cgcctgctgc tgcccggcga gctggccaag cacgccgtgt ccgagggcac caaggcggtc
      661 accaagtaca ccagctccaa gggatcgggt gggactagtg gcgtgagcaa gggcgaggag
      721 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
      781 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
      841 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
      901 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
      961 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     1021 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     1081 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     1141 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     1201 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     1261 gccaagaagc ccgtgcagct gcccggcgcc gtaagtatga aattcaggga tacggccacc
     1321 ttgttggcgg gccagatata cgcgttgacg agtagtcaga ataagcggag gccctgaggg
     1381 gcaccatacc gagtgatggg gcccatcagc taccataggc accacgagcg gcaggggcta
     1441 attcactccc aacgagcaag agtagtcaga ataagcggag gccctgaggg gcaccatacc
     1501 gagtgatggg gcccatcagc taccataggc accacgagcg gcagccttga tctgtggatc
     1561 taccagaacg agtagtcaga ataagcggag gccctgaggg gcaccatacc gagtgatggg
     1621 gcccatcagc taccataggc accacgagcg gcagggcaga actacacacc agggccagtg
     1681 agtagtcaga ataagcggag gccctgaggg gcaccatacc gagtgatggg gcccatcagc
     1741 taccataggc accacgagcg gcagcatcgg catctttgac gcaagacacg agtagtcaga
     1801 ataagcggag gccctgaggg gcaccatacc gagtgatggg gcccatcagc taccataggc
     1861 accacgagcg gcagccgctt gttacaccct gtgagtgtcg agtagtcaga ataagcggag
     1921 gccctgaggg gcaccatacc gagtgatggg gcccatcagc taccataggc accacgagcg
     1981 gcaggcagta gtatctttgc aagtgactca cccaccttgt tggcatattt gccaaatagt
     2041 ggaaatgtga agtactgaca aaacttttcc ctttttcaat ctaatagtac aacgtcaaca
     2101 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     2161 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagggaacc ggtgctggaa
     2221 gtggttgagt acctgccggg acctggcgct cgctcgctcg agtcgccggc tgcttgactc
     2281 caaaggctct tttcagagcc acccacctaa tcactagaaa agagcttgtt cacttattcc
     2341 cttagtttct tttcataaag taagttattt tagtgtgaag gtcatgggaa atggcatacg
     2401 tagcttttta actatttgga actcgaggtc cccagtgcgt cattggattt gcttttgaat
     2461 ctagagcgtg tctttactca ttgtgctgct tagccttccc aggagtcggt tctcaattag
     2521 gctgttggga atccgcctct ttacccgccc ccactcccgc cccacacgcg ccctggtggc
     2581 tccttgggtc tgtttcattc taaaacgaag tggctgagtt cggctgtcat ttgcgagcgc
     2641 caggtcccgg cagggcccgg gcgacggatg gtgatccccc tggccagtgc acgtctgctg
     2701 tcagataaag tctcccgtga actttacccg gtggtgcata tcggggatga aagctggcgc
     2761 atgatgacca ccgatatggc cagtgtgccg gtctccgtta tcggggaaga agtggctgat
     2821 ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa
     2881 atgtcaggct cccttataca cagccagtct gcaggtcgac ggtctcgact cacatgtgag
     2941 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3001 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3061 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3121 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3181 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3241 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3301 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3361 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3421 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3481 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3541 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3601 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3661 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3721 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3781 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3841 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagaaccac
     3901 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3961 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4021 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4081 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4141 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4201 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4261 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4321 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4381 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4441 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4501 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4561 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4621 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4681 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4741 ctgacgtc
//
