LOCUS Exported 3896 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses Ollas::mCherry::linker::H2B::V5 in bacteria. ACCESSION . VERSION . KEYWORDS AP582-2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3896) AUTHORS Paix A, Folkmann A, Rasoloson D, Seydoux G TITLE High Efficiency, Homology-Directed Genome Editing in Caenorhabditis elegans Using CRISPR-Cas9 Ribonucleoprotein Complexes. JOURNAL Genetics. 2015 Sep;201(1):47-54. doi: 10.1534/genetics.115.179382. Epub 2015 Jul 17. PUBMED 26187122 REFERENCE 2 (bases 1 to 3896) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3896 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 106..127 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 142..172 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 180..196 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 185..207 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 204..220 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 204..220 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS complement(268..309) /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS complement(1432..1473) /codon_start=1 /product="Escherichia coli OmpF Linker and mouse Langerin fusion sequence tag (Park et al., 2008)" /label=OLLAS tag /translation="SGFANELGPRLMGK" primer_bind complement(1500..1517) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(1500..1516) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1509..1531) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(1725..1744) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1844..1866 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(1904..1922) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1990..2094 /gene="bla" /label=AmpR promoter CDS 2095..2955 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(2313..2332) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3126..3714 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3615..3634 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3868..3885 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct 121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat 181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc 241 atgcctgcag gtcgactcta gaggatcagt actatctaga ccgaggagag ggttagggat 301 aggcttaccc ttgctggaag tgtacttggt gacggccttg gttccctcag acacggcgtg 361 cttggcaagt tctcctggga gaatcaaacg gacagcggtt tgaatttcgc gggatgagat 421 cgttgagcgt ttgttgtaat gagcaagacg ggaagcttcc gaagcgatgc gttcgaatac 481 atcgttgacg aaggagttca tgatagacat ggccttggag gagactccgg tgtctgggtg 541 aacttgcttg agaacacggt agatgtagac ggagtacgat tccttgcggg catgacgtct 601 cttctttccg tcctttggct tggcaacgac ggtcttggcg gccttcttgg ctcccttggc 661 agatggcttt ggtggactag ttctagagcg gccgccaccg gtggagctcc accgatcccc 721 cgggcactta tacaattcat ccatgccacc tgtcgagtgc cgcccttcag ctctttcata 781 ttgctcaacg atagtgtaat cctcgttgtg ggaggtgata tccagtttaa tattaacatt 841 gtacgctcct ggcagctgaa ctggtttttt ggctttgtaa gttgtcttca cctcagcatc 901 gtaatgtcct ccatctttca atttaagacg ttgtttaatc tctcccttaa gagcaccgtc 961 ttcagggtac attctttcac ttgaagcttc ccatcccata gtcttcttct gcataacagg 1021 tccatccgag gggaaattcg ttcctcgaag tttgacttta taaataaatt ctccatcttg 1081 gaggctggag tcttgcgtca ctgtaacaac tcctccgtct tcaaaattca ttacgcgttc 1141 ccacttaaaa ccctccggga atgaaagttt caaataatct ggtatatctg ccggatgttt 1201 aacataagct ttacttccat acatgaactg tggagagagg atgtcccagg cgaatggtaa 1261 tggtccgcct ttagttacct ttagttttgc ggtttgtgtt ccctcatatg gtcggccttc 1321 tccttcacct tcaatttcaa actcatgccc attgacagat ccctccatat gcaccttgaa 1381 acgcataaac tctttaataa ttgccatgtt atcttcttca ccctttgaga cctttcccat 1441 gagacgtggt ccgagctcgt tggcgaatcc ggagatcccc gggtaccgag ctcgaattca 1501 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc 1561 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc 1621 ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt 1681 acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat 1741 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct 1801 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt 1861 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta 1921 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg 1981 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg 2041 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt 2101 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt 2161 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg 2221 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa 2281 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt 2341 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag 2401 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt 2461 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga 2521 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt 2581 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta 2641 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg 2701 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc 2761 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt 2821 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg 2881 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg 2941 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa 3001 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa 3061 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga 3121 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg 3181 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact 3241 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac 3301 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg 3361 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg 3421 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga 3481 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc 3541 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg 3601 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc 3661 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc 3721 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt 3781 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc 3841 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaaga //