LOCUS Exported 2860 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION SapTrap SL2::SV40-NLS C-terminal connector donor plasmid. ACCESSION . VERSION . KEYWORDS pMLS269 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2860) AUTHORS Schwartz ML, Jorgensen EM TITLE SapTrap, a Toolkit for High-Throughput CRISPR/Cas9 Gene Modification in Caenorhabditis elegans. JOURNAL Genetics. 2016 Feb 2. pii: genetics.115.184275. PUBMED 26837755 REFERENCE 2 (bases 1 to 2860) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2860 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 92..109 /label=L4440 /note="L4440 vector, forward primer" terminator 268..295 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 387..473 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 522..544 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 536..553 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 537..553 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 569..668 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 940..960 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" protein_bind complement(985..1084) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1101..1120) /label=T7 /note="T7 promoter, forward primer" promoter complement(1102..1120) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1125..1141) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1125..1141) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1243..1262) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1254..2063 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(1321..1340) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2210..2798 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2699..2718 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga 61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga 121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc 241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta 301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc 361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa 421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg 481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa 541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac 601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa 661 agcaggctat gctcttcggc gtaagctgtc tcatcctact ttcacctagt taactgcttg 721 tcttaaaatc tatgcttctc tttagtatct aaaattttcc tagaagctta caagtatata 781 aatggtctct tctcaataaa ggttgtatat ttattcatct tattgaatct gccatttcct 841 cgtttttgcg agtttatata ccttccaatt ttctttctat tgtattttca acttctaatt 901 ttaattcagg gaaactgctt caacgcatca atgactgctc caaagaagaa gcgtaaggta 961 ccggtagaaa aaatgcgaag agctacccag ctttcttgta caaagttggc attataagaa 1021 agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta 1081 tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct 1141 ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaataatat 1201 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc 1261 atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg 1321 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg 1381 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg 1441 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca 1501 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa 1561 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg 1621 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc 1681 gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg 1741 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac 1801 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta 1861 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc 1921 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga 1981 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt 2041 tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca 2101 ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac gtgagttacg 2161 cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt 2221 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt 2281 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc 2341 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg 2401 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg 2461 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt 2521 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac 2581 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg 2641 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg 2701 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat 2761 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt 2821 tacggttcct ggccttttgc tggccttttg ctcacatgtt //