LOCUS       sequence-392210-        4823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR-Tag sequence that can be efficiently bound by dSpCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4823)
  AUTHORS   Xu H, Wang J, Deng Y, Hou F, Fu Y, Chen S, Zou W, Pan D, Chen B
  TITLE     Two-Color CRISPR Imaging Reveals Dynamics of Herpes Simplex Virus 1 
            Replication Compartments and Virus-Host Interactions.
  JOURNAL   J Virol. 2022 Dec 21;96(24):e0092022. doi: 10.1128/jvi.00920-22. 
            Epub 2022 Dec 1.
  PUBMED    36453882
REFERENCE   2  (bases 1 to 4823)
  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: "J Virol."; 
            date: "2022-12-21"; volume: "96(24):e0092022. doi"; pages: " 
            10.1128/jvi.00920-22. Epub 2022 Dec 1"
COMMENT     Sequence Label: pCA17-CRISPR-Tag (11XTS1) for dSpCas9
FEATURES             Location/Qualifiers
     source          1..4823
                     /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             703..1410
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     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     2834..2853
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     rep_origin      complement(3068..3656)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3148..3167)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3827..4618,4619..4687))
                     /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     4450..4469
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4688..4792)
                     /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 tacaacgtca acatcaagtt ggacatcacc
     1321 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     1381 accggcggca tggacgagct gtacaagtga ggatccacgc gttgacaatc tgacaaaggt
     1441 ggcgtacctc gcaccgatgc tctccgagga ggcacctacg tacgcaccga tgctctccga
     1501 ggcagcaacg acatcggagt agtcgcaccg atgctctccg aggaggcgta cacggttgga
     1561 tcataggata gcaaaatctg acaaaggtgg cgtacctcgc accgatgctc tccgaggagg
     1621 cacctacgta cgcaccgatg ctctccgagg cagcaacgac atcggagtag tcgcaccgat
     1681 gctctccgag gaggcgtaca cggttggatc ataggatagg cagaactaca caccagggcg
     1741 aacaatctga caaaggtggc gtacctcgca ccgatgctct ccgaggaggc gtacacggtt
     1801 ggatcatagg atacagtaat ctgacaaagg tggcgtacct cgcaccgatg ctctccgagg
     1861 aggcacctac gtacgcaccg atgctctccg aggcagcaac gacatcggag tagtcgcacc
     1921 gatgctctcc gaggaggcgt acacggttgg atcataggat aacacaatct gacaaaggtg
     1981 gcgtacctcg caccgatgct ctccgaggag gcacctacgt acgcaccgat gctctccgag
     2041 gcagcaacga catcggagta gtcgcaccga tgctctccga ggaggcgtac acggttggat
     2101 cataggatat gtcaatctga caaaggtggc gtacctcgca ccgatgctct ccgaggaggc
     2161 acctacgtac gcaccgatgc tctccgaggc agcaacgaca tcggagtagt cgcaccgatg
     2221 ctctccgagg aggcgtacac ggttggatca taggataact cacatgtgag caaaaggccg
     2281 gaaccggtgc tggaagtggt tgagtacctg ccgggacctg gcgctcgctc gctcgagtcg
     2341 ccggctgctt gactccaaag gctcttttca gagccaccca cctaatcact agaaaagagc
     2401 ttgttcactt attcccttag tttcttttca taaagtaagt tattttagtg tgaaggtcat
     2461 gggaaatggc atacgtagct ttttaactat ttggaactcg aggtccccag tgcgtcattg
     2521 gatttgcttt tgaatctaga gcgtgtcttt actcattgtg ctgcttagcc ttcccaggag
     2581 tcggttctca attaggctgt tgggaatccg cctctttacc cgcccccact cccgccccac
     2641 acgcgccctg gtggctcctt gggtctgttt cattctaaaa cgaagtggct gagttcggct
     2701 gtcatttgcg agcgccaggt cccggcaggg cccgggcgac ggatggtgat ccccctggcc
     2761 agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt acccggtggt gcatatcggg
     2821 gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtctc cgttatcggg
     2881 gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat taacctgatg
     2941 ttctggggaa tataaatgtc aggctccctt atacacagcc agtctgcagg tcgacggtct
     3001 cgactcacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3061 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3121 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3181 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3241 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3301 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3361 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3421 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3481 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3541 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3601 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3661 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3721 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3781 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3841 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3901 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3961 taccgcgaga accacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4021 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4081 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4141 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4201 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4261 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4321 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4381 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4441 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4501 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4561 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4621 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4681 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4741 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4801 cccgaaaagt gccacctgac gtc
//
