LOCUS Exported 3550 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPaint gene tagging. ACCESSION . VERSION . KEYWORDS pCRISPaint-nTEV-PuroR SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3550) 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 3550) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3550 /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 581..634 /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 1258..1379 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1269..1288) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1323..1342 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(1627..1644) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1798..2386) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1878..1897) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2586..3446) /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 gagagcctgt tcaagggacc 241 cagagactac aaccccatca gcagcaccat ctgccacctg accaacgaga gcgacggcca 301 caccaccagc ctgtacggca tcggcttcgg ccccttcatc atcaccaaca agcacctgtt 361 cagaagaaac aacggcaccc tgctggtgca gagcctgcac ggcgtgttca aggtgaagaa 421 caccaccacc ctgcagcagc acctgatcga cggcagagac atgatcatca tcagaatgcc 481 caaggacttc ccccccttcc cccagaagct gaagttcaga gagccccaga gagaggagag 541 aatctgcctg gtgaccacca acttccagac ctccggagga gagggcagag gaagtctgct 601 aacatgcggt gacgtcgagg agaatcctgg cccaatgact gaatacaagc ctactgtcag 661 gttggctaca agagacgacg ttcctagagc cgtgagaact ctggctgcag ccttcgccga 721 ctaccccgcc acgagacaca ccgttgaccc agatcggcat attgagagag tgactgaact 781 gcaggagctg tttcttacaa gagttggcct cgacataggc aaggtgtggg tggcggacga 841 cggcgccgcc gtggccgtct ggaccactcc cgaatcagtt gaggctggcg ccgtattcgc 901 tgagatcggc ccgagaatgg ctgagctcag cgggagtagg ctcgcggcac agcagcaaat 961 ggagggactg ctggcaccac acaggcccaa agaacccgcc tggttcctgg caaccgtcgg 1021 tgtatctccc gatcatcagg ggaaaggtct gggctctgcc gtagtgctcc ctggcgtgga 1081 ggcagctgag agagcaggag tacctgcctt cttggagacc tccgctccaa ggaatcttcc 1141 cttctatgaa cggttgggct tcaccgtgac agccgacgtg gaagtccccg aaggcccccg 1201 cacttggtgc atgacgagga agcctggagc gtgagggccc gcttcgagca gacatgataa 1261 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt 1321 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta 1381 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt 1441 aaagcaagta aaacctctac aaatgtggta aaatcgataa agatccgtcg accgatgccc 1501 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc 1561 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc 1621 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc 1681 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat 1741 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt 1801 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg 1861 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc 1921 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt 1981 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa 2041 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta 2101 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa 2161 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa 2221 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt 2281 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt 2341 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat 2401 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat 2461 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc 2521 aatctaaagt atatatgagt aaacttggtc tgacagcggc cgcaaatgct aaaccactgc 2581 agtggttacc agtgcttgat cagtgaggca ccgatctcag cgatctgcct atttcgttcg 2641 tccatagtgg cctgactccc cgtcgtgtag atcactacga ttcgtgaggg cttaccatca 2701 ggccccagcg cagcaatgat gccgcgagag ccgcgttcac cggcccccga tttgtcagca 2761 atgaaccagc cagcagggag ggccgagcga agaagtggtc ctgctacttt gtccgcctcc 2821 atccagtcta tgagctgctg tcgtgatgct agagtaagaa gttcgccagt gagtagtttc 2881 cgaagagttg tggccattgc tactggcatc gtggtatcac gctcgtcgtt cggtatggct 2941 tcgttcaact ctggttccca gcggtcaagc cgggtcacat gatcacccat attatgaaga 3001 aatgcagtca gctccttagg gcctccgatc gttgtcagaa gtaagttggc cgcggtgttg 3061 tcgctcatgg taatggcagc actacacaat tctcttaccg tcatgccatc cgtaagatgc 3121 ttttccgtga ccggcgagta ctcaaccaag tcgttttgtg agtagtgtat acggcgacca 3181 agctgctctt gcccggcgtc tatacgggac aacaccgcgc cacatagcag tactttgaaa 3241 gtgctcatca tcgggaatcg ttcttcgggg cggaaagact caaggatctt gccgctattg 3301 agatccagtt cgatatagcc cactcttgca cccagttgat cttcagcatc ttttactttc 3361 accagcgttt cggggtgtgc aaaaacaggc aagcaaaatg ccgcaaagaa gggaatgagt 3421 gcgacacgaa aatgttggat gctcatactc gtcctttttc aatattattg aagcatttat 3481 cagggttact agtacgtctc tcaaggataa gtaagtaata ttaaggtacg ggaggtattg 3541 gacaggccgc //