LOCUS Exported 3895 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION CRISPaint gene tagging. ACCESSION . VERSION . KEYWORDS pCRISPaint-TagBFP-PuroR SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3895) 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 3895) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3895 /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..916 /codon_start=1 /product="monomeric blue fluorescent protein" /label=TagBFP /note="mammalian codon-optimized" /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC DLPSKLGHKLN" CDS 926..979 /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 1603..1724 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1614..1633) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1668..1687 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(1972..1989) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2143..2731) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2223..2242) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2931..3791) /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 gggatccatg agcgagctga ttaaggagaa 241 catgcacatg aagctgtaca tggagggcac cgtggacaac catcacttca agtgcacatc 301 cgagggcgaa ggcaagccct acgagggcac ccagaccatg agaatcaagg tggtcgaggg 361 cggccctctc cccttcgcct tcgacatcct ggctactagc ttcctctacg gcagcaagac 421 cttcatcaac cacacccagg gcatccccga cttcttcaag cagtccttcc ctgagggctt 481 cacatgggag agagtcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac 541 cagcctccag gacggctgcc tcatctacaa cgtcaagatc agaggggtga acttcacatc 601 caacggccct gtgatgcaga agaaaacact cggctgggag gccttcaccg agacgctgta 661 ccccgctgac ggcggcctgg aaggcagaaa cgacatggcc ctgaagctcg tgggcgggag 721 ccatctgatc gcaaacatca agaccacata tagatccaag aaacccgcta agaacctcaa 781 gatgcctggc gtctactatg tggactacag actggaaaga atcaaggagg ccaacaacga 841 gacctacgtc gagcagcacg aggtggcagt ggccagatac tgcgacctcc ctagcaaact 901 ggggcacaag cttaattccg gaggagaggg cagaggaagt ctgctaacat gcggtgacgt 961 cgaggagaat cctggcccaa tgactgaata caagcctact gtcaggttgg ctacaagaga 1021 cgacgttcct agagccgtga gaactctggc tgcagccttc gccgactacc ccgccacgag 1081 acacaccgtt gacccagatc ggcatattga gagagtgact gaactgcagg agctgtttct 1141 tacaagagtt ggcctcgaca taggcaaggt gtgggtggcg gacgacggcg ccgccgtggc 1201 cgtctggacc actcccgaat cagttgaggc tggcgccgta ttcgctgaga tcggcccgag 1261 aatggctgag ctcagcggga gtaggctcgc ggcacagcag caaatggagg gactgctggc 1321 accacacagg cccaaagaac ccgcctggtt cctggcaacc gtcggtgtat ctcccgatca 1381 tcaggggaaa ggtctgggct ctgccgtagt gctccctggc gtggaggcag ctgagagagc 1441 aggagtacct gccttcttgg agacctccgc tccaaggaat cttcccttct atgaacggtt 1501 gggcttcacc gtgacagccg acgtggaagt ccccgaaggc ccccgcactt ggtgcatgac 1561 gaggaagcct ggagcgtgag ggcccgcttc gagcagacat gataagatac attgatgagt 1621 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg 1681 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca 1741 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc 1801 tctacaaatg tggtaaaatc gataaagatc cgtcgaccga tgcccttgag agccttcaac 1861 ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc 1921 ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt cctcgctcac 1981 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 2041 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 2101 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 2161 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 2221 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 2281 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 2341 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 2401 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 2461 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 2521 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 2581 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 2641 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 2701 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 2761 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 2821 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 2881 tgagtaaact tggtctgaca gcggccgcaa atgctaaacc actgcagtgg ttaccagtgc 2941 ttgatcagtg aggcaccgat ctcagcgatc tgcctatttc gttcgtccat agtggcctga 3001 ctccccgtcg tgtagatcac tacgattcgt gagggcttac catcaggccc cagcgcagca 3061 atgatgccgc gagagccgcg ttcaccggcc cccgatttgt cagcaatgaa ccagccagca 3121 gggagggccg agcgaagaag tggtcctgct actttgtccg cctccatcca gtctatgagc 3181 tgctgtcgtg atgctagagt aagaagttcg ccagtgagta gtttccgaag agttgtggcc 3241 attgctactg gcatcgtggt atcacgctcg tcgttcggta tggcttcgtt caactctggt 3301 tcccagcggt caagccgggt cacatgatca cccatattat gaagaaatgc agtcagctcc 3361 ttagggcctc cgatcgttgt cagaagtaag ttggccgcgg tgttgtcgct catggtaatg 3421 gcagcactac acaattctct taccgtcatg ccatccgtaa gatgcttttc cgtgaccggc 3481 gagtactcaa ccaagtcgtt ttgtgagtag tgtatacggc gaccaagctg ctcttgcccg 3541 gcgtctatac gggacaacac cgcgccacat agcagtactt tgaaagtgct catcatcggg 3601 aatcgttctt cggggcggaa agactcaagg atcttgccgc tattgagatc cagttcgata 3661 tagcccactc ttgcacccag ttgatcttca gcatctttta ctttcaccag cgtttcgggg 3721 tgtgcaaaaa caggcaagca aaatgccgca aagaagggaa tgagtgcgac acgaaaatgt 3781 tggatgctca tactcgtcct ttttcaatat tattgaagca tttatcaggg ttactagtac 3841 gtctctcaag gataagtaag taatattaag gtacgggagg tattggacag gccgc //