LOCUS       sequence-153089-        3241 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 3241)
  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 3241)
  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-AviTag-PuroR
FEATURES             Location/Qualifiers
     source          1..3241
                     /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..262
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             272..325
                     /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    949..1070
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(960..979)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1014..1033
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1318..1335)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1489..2077)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1569..1588)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2277..3068,3069..3137))
                     /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 gggatccggc ctgaacgaca tcttcgaggc
      241 ccagaagatc gagtggcacg agtccggagg agagggcaga ggaagtctgc taacatgcgg
      301 tgacgtcgag gagaatcctg gcccaatgac tgaatacaag cctactgtca ggttggctac
      361 aagagacgac gttcctagag ccgtgagaac tctggctgca gccttcgccg actaccccgc
      421 cacgagacac accgttgacc cagatcggca tattgagaga gtgactgaac tgcaggagct
      481 gtttcttaca agagttggcc tcgacatagg caaggtgtgg gtggcggacg acggcgccgc
      541 cgtggccgtc tggaccactc ccgaatcagt tgaggctggc gccgtattcg ctgagatcgg
      601 cccgagaatg gctgagctca gcgggagtag gctcgcggca cagcagcaaa tggagggact
      661 gctggcacca cacaggccca aagaacccgc ctggttcctg gcaaccgtcg gtgtatctcc
      721 cgatcatcag gggaaaggtc tgggctctgc cgtagtgctc cctggcgtgg aggcagctga
      781 gagagcagga gtacctgcct tcttggagac ctccgctcca aggaatcttc ccttctatga
      841 acggttgggc ttcaccgtga cagccgacgt ggaagtcccc gaaggccccc gcacttggtg
      901 catgacgagg aagcctggag cgtgagggcc cgcttcgagc agacatgata agatacattg
      961 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1021 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1081 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     1141 aaaacctcta caaatgtggt aaaatcgata aagatccgtc gaccgatgcc cttgagagcc
     1201 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
     1261 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctt ccgcttcctc
     1321 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1381 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1441 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1501 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1561 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1621 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1681 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     1741 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     1801 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     1861 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     1921 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     1981 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2041 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2101 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2161 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2221 tatatatgag taaacttggt ctgacagcgg ccgcaaatgc taaaccactg cagtggttac
     2281 cagtgcttga tcagtgaggc accgatctca gcgatctgcc tatttcgttc gtccatagtg
     2341 gcctgactcc ccgtcgtgta gatcactacg attcgtgagg gcttaccatc aggccccagc
     2401 gcagcaatga tgccgcgaga gccgcgttca ccggcccccg atttgtcagc aatgaaccag
     2461 ccagcaggga gggccgagcg aagaagtggt cctgctactt tgtccgcctc catccagtct
     2521 atgagctgct gtcgtgatgc tagagtaaga agttcgccag tgagtagttt ccgaagagtt
     2581 gtggccattg ctactggcat cgtggtatca cgctcgtcgt tcggtatggc ttcgttcaac
     2641 tctggttccc agcggtcaag ccgggtcaca tgatcaccca tattatgaag aaatgcagtc
     2701 agctccttag ggcctccgat cgttgtcaga agtaagttgg ccgcggtgtt gtcgctcatg
     2761 gtaatggcag cactacacaa ttctcttacc gtcatgccat ccgtaagatg cttttccgtg
     2821 accggcgagt actcaaccaa gtcgttttgt gagtagtgta tacggcgacc aagctgctct
     2881 tgcccggcgt ctatacggga caacaccgcg ccacatagca gtactttgaa agtgctcatc
     2941 atcgggaatc gttcttcggg gcggaaagac tcaaggatct tgccgctatt gagatccagt
     3001 tcgatatagc ccactcttgc acccagttga tcttcagcat cttttacttt caccagcgtt
     3061 tcggggtgtg caaaaacagg caagcaaaat gccgcaaaga agggaatgag tgcgacacga
     3121 aaatgttgga tgctcatact cgtccttttt caatattatt gaagcattta tcagggttac
     3181 tagtacgtct ctcaaggata agtaagtaat attaaggtac gggaggtatt ggacaggccg
     3241 c
//
