LOCUS Exported 3214 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPaint gene tagging. ACCESSION . VERSION . KEYWORDS pCRISPaint-TC-PuroR SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3214) 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 3214) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3214 /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..235 /codon_start=1 /product="tetracysteine peptide that binds biarsenical labeling reagents" /label=tetracysteine tag /translation="CCPGCC" CDS 245..298 /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 922..1043 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(933..952) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 987..1006 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(1291..1308) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1462..2050) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1542..1561) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2250..3110) /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 gggatcctgc tgccccggct gctgctccgg 241 aggagagggc agaggaagtc tgctaacatg cggtgacgtc gaggagaatc ctggcccaat 301 gactgaatac aagcctactg tcaggttggc tacaagagac gacgttccta gagccgtgag 361 aactctggct gcagccttcg ccgactaccc cgccacgaga cacaccgttg acccagatcg 421 gcatattgag agagtgactg aactgcagga gctgtttctt acaagagttg gcctcgacat 481 aggcaaggtg tgggtggcgg acgacggcgc cgccgtggcc gtctggacca ctcccgaatc 541 agttgaggct ggcgccgtat tcgctgagat cggcccgaga atggctgagc tcagcgggag 601 taggctcgcg gcacagcagc aaatggaggg actgctggca ccacacaggc ccaaagaacc 661 cgcctggttc ctggcaaccg tcggtgtatc tcccgatcat caggggaaag gtctgggctc 721 tgccgtagtg ctccctggcg tggaggcagc tgagagagca ggagtacctg ccttcttgga 781 gacctccgct ccaaggaatc ttcccttcta tgaacggttg ggcttcaccg tgacagccga 841 cgtggaagtc cccgaaggcc cccgcacttg gtgcatgacg aggaagcctg gagcgtgagg 901 gcccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa 961 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca 1021 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc 1081 agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg 1141 ataaagatcc gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg 1201 gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta 1261 ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 1321 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 1381 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 1441 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 1501 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 1561 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 1621 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 1681 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 1741 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 1801 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 1861 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 1921 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 1981 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 2041 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 2101 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 2161 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag 2221 cggccgcaaa tgctaaacca ctgcagtggt taccagtgct tgatcagtga ggcaccgatc 2281 tcagcgatct gcctatttcg ttcgtccata gtggcctgac tccccgtcgt gtagatcact 2341 acgattcgtg agggcttacc atcaggcccc agcgcagcaa tgatgccgcg agagccgcgt 2401 tcaccggccc ccgatttgtc agcaatgaac cagccagcag ggagggccga gcgaagaagt 2461 ggtcctgcta ctttgtccgc ctccatccag tctatgagct gctgtcgtga tgctagagta 2521 agaagttcgc cagtgagtag tttccgaaga gttgtggcca ttgctactgg catcgtggta 2581 tcacgctcgt cgttcggtat ggcttcgttc aactctggtt cccagcggtc aagccgggtc 2641 acatgatcac ccatattatg aagaaatgca gtcagctcct tagggcctcc gatcgttgtc 2701 agaagtaagt tggccgcggt gttgtcgctc atggtaatgg cagcactaca caattctctt 2761 accgtcatgc catccgtaag atgcttttcc gtgaccggcg agtactcaac caagtcgttt 2821 tgtgagtagt gtatacggcg accaagctgc tcttgcccgg cgtctatacg ggacaacacc 2881 gcgccacata gcagtacttt gaaagtgctc atcatcggga atcgttcttc ggggcggaaa 2941 gactcaagga tcttgccgct attgagatcc agttcgatat agcccactct tgcacccagt 3001 tgatcttcag catcttttac tttcaccagc gtttcggggt gtgcaaaaac aggcaagcaa 3061 aatgccgcaa agaagggaat gagtgcgaca cgaaaatgtt ggatgctcat actcgtcctt 3121 tttcaatatt attgaagcat ttatcagggt tactagtacg tctctcaagg ataagtaagt 3181 aatattaagg tacgggaggt attggacagg ccgc //