LOCUS       sequence-153091-        3223 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPaint gene tagging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3223)
  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 3223)
  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-HA-PuroR
FEATURES             Location/Qualifiers
     source          1..3223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1..49
                     /note="synthetic polyadenylation signal"
     misc_feature    63..154
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     103..122
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     CDS             218..244
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             218..244
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             254..307
                     /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"
     polyA_signal    931..1052
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(942..961)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     996..1015
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1300..1317)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1471..2059)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1551..1570)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2259..3050,3051..3119))
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga atcgatagta
       61 ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa ataggctgtc
      121 cccagtgcaa gtgcaggtgc cagaacattt ctcttaatga tagctagcgg gccagtaccc
      181 aaaaagcggg gggtctggtg gcagtggagg gggatcctac ccctacgacg tgcccgacta
      241 cgcctccgga ggagagggca gaggaagtct gctaacatgc ggtgacgtcg aggagaatcc
      301 tggcccaatg actgaataca agcctactgt caggttggct acaagagacg acgttcctag
      361 agccgtgaga actctggctg cagccttcgc cgactacccc gccacgagac acaccgttga
      421 cccagatcgg catattgaga gagtgactga actgcaggag ctgtttctta caagagttgg
      481 cctcgacata ggcaaggtgt gggtggcgga cgacggcgcc gccgtggccg tctggaccac
      541 tcccgaatca gttgaggctg gcgccgtatt cgctgagatc ggcccgagaa tggctgagct
      601 cagcgggagt aggctcgcgg cacagcagca aatggaggga ctgctggcac cacacaggcc
      661 caaagaaccc gcctggttcc tggcaaccgt cggtgtatct cccgatcatc aggggaaagg
      721 tctgggctct gccgtagtgc tccctggcgt ggaggcagct gagagagcag gagtacctgc
      781 cttcttggag acctccgctc caaggaatct tcccttctat gaacggttgg gcttcaccgt
      841 gacagccgac gtggaagtcc ccgaaggccc ccgcacttgg tgcatgacga ggaagcctgg
      901 agcgtgaggg cccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
      961 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     1021 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     1081 ttcaggttca gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     1141 gtaaaatcga taaagatccg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc
     1201 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg
     1261 caactcgtag gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg
     1321 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     1381 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     1441 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1501 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1561 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1621 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1681 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1741 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1801 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     1861 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     1921 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1981 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2041 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2101 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2161 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2221 gtctgacagc ggccgcaaat gctaaaccac tgcagtggtt accagtgctt gatcagtgag
     2281 gcaccgatct cagcgatctg cctatttcgt tcgtccatag tggcctgact ccccgtcgtg
     2341 tagatcacta cgattcgtga gggcttacca tcaggcccca gcgcagcaat gatgccgcga
     2401 gagccgcgtt caccggcccc cgatttgtca gcaatgaacc agccagcagg gagggccgag
     2461 cgaagaagtg gtcctgctac tttgtccgcc tccatccagt ctatgagctg ctgtcgtgat
     2521 gctagagtaa gaagttcgcc agtgagtagt ttccgaagag ttgtggccat tgctactggc
     2581 atcgtggtat cacgctcgtc gttcggtatg gcttcgttca actctggttc ccagcggtca
     2641 agccgggtca catgatcacc catattatga agaaatgcag tcagctcctt agggcctccg
     2701 atcgttgtca gaagtaagtt ggccgcggtg ttgtcgctca tggtaatggc agcactacac
     2761 aattctctta ccgtcatgcc atccgtaaga tgcttttccg tgaccggcga gtactcaacc
     2821 aagtcgtttt gtgagtagtg tatacggcga ccaagctgct cttgcccggc gtctatacgg
     2881 gacaacaccg cgccacatag cagtactttg aaagtgctca tcatcgggaa tcgttcttcg
     2941 gggcggaaag actcaaggat cttgccgcta ttgagatcca gttcgatata gcccactctt
     3001 gcacccagtt gatcttcagc atcttttact ttcaccagcg tttcggggtg tgcaaaaaca
     3061 ggcaagcaaa atgccgcaaa gaagggaatg agtgcgacac gaaaatgttg gatgctcata
     3121 ctcgtccttt ttcaatatta ttgaagcatt tatcagggtt actagtacgt ctctcaagga
     3181 taagtaagta atattaaggt acgggaggta ttggacaggc cgc
//
