LOCUS       sequence-158415-         952 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  CRISPaint gene tagging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 952)
  AUTHORS   Schmid-Burgk JL, Honing K, Ebert TS, Hornung V
  TITLE     CRISPaint allows modular base-specific gene tagging using a 
            ligase-4-dependent mechanism.
  JOURNAL   Nat Commun. 2016 Jul 28;7:12338. doi: 10.1038/ncomms12338.
  PUBMED    27465542
REFERENCE   2  (bases 1 to 952)
  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: "2016-07-28"; volume: "7:12338. doi"; pages: " 
            10.1038/ncomms12338"
COMMENT     Sequence Label: pCRISPaint-SpyTag-PuroR
FEATURES             Location/Qualifiers
     source          1..952
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(609..662)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(672..710)
                     /codon_start=1
                     /product="peptide derived from the CnaB2 adhesin domain of 
                     Streptococcus pyogenes FbaB"
                     /label=SpyTag
                     /note="forms a covalent isopeptide bond with the SpyCatcher
                     protein (Zakeri et al., 2012)"
                     /translation="AHIVMVDAYKPTK"
     misc_feature    774..865
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(806..825)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    879..927
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 gcgggccctc acgctccagg cttcctcgtc atgcaccaag tgcgggggcc ttcggggact
       61 tccacgtcgg ctgtcacggt gaagcccaac cgttcataga agggaagatt ccttggagcg
      121 gaggtctcca agaaggcagg tactcctgct ctctcagctg cctccacgcc agggagcact
      181 acggcagagc ccagaccttt cccctgatga tcgggagata caccgacggt tgccaggaac
      241 caggcgggtt ctttgggcct gtgtggtgcc agcagtccct ccatttgctg ctgtgccgcg
      301 agcctactcc cgctgagctc agccattctc gggccgatct cagcgaatac ggcgccagcc
      361 tcaactgatt cgggagtggt ccagacggcc acggcggcgc cgtcgtccgc cacccacacc
      421 ttgcctatgt cgaggccaac tcttgtaaga aacagctcct gcagttcagt cactctctca
      481 atatgccgat ctgggtcaac ggtgtgtctc gtggcggggt agtcggcgaa ggctgcagcc
      541 agagttctca cggctctagg aacgtcgtct cttgtagcca acctgacagt aggcttgtat
      601 tcagtcattg ggccaggatt ctcctcgacg tcaccgcatg ttagcagact tcctctgccc
      661 tctcctccgg acttggtggg cttgtaggcg tccaccatca cgatgtgggc ggatccccct
      721 ccactgccac cagacccccc gctttttggg tactggcccg ctagctatca ttaagagaaa
      781 tgttctggca cctgcacttg cactggggac agcctatttt gctagtttgt tttgtttcgt
      841 tttgttttga tggagagcgt atgttagtac tatcgattca cacaaaaaac caacacacag
      901 atgtaatgaa aataaagata ttttattgcg gcctgtccaa tacctcccgt ac
//
