LOCUS Exported 3241 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION CRISPaint gene tagging. ACCESSION . VERSION . KEYWORDS pCRISPaint-AviTag-PuroR 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3241 /organism="synthetic DNA construct" /mol_type="other DNA" 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(2277..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 //