LOCUS Exported 3223 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION CRISPaint gene tagging. ACCESSION . VERSION . KEYWORDS pCRISPaint-HA-PuroR 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3223 /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..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(2259..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 //