LOCUS Exported 3159 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION SapTrap syntron-embedded FLP-on C-terminal connector donor plasmid. ACCESSION . VERSION . KEYWORDS pMLS272 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3159) 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 3159) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3159 /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" protein_bind 706..739 /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" protein_bind 1178..1211 /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" protein_bind complement(1284..1383) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1400..1419) /label=T7 /note="T7 promoter, forward primer" promoter complement(1401..1419) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1424..1440) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1424..1440) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1542..1561) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1553..2362 /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(1620..1639) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2509..3097 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2998..3017 /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 agcaggctgc tcttcggcgg taagtttaaa catgatttta ctaacgaagt tcctattctc 721 tagaaagtat aggaacttct aactaatctg atttaaattt tcagggttaa attttcaaat 781 tttaaatact gaatatttgt tttttttcct attatttatt tattctcttt gtgttttttt 841 tcttgctttc taaaaaatta attcaatcca aatctaaaca tttttttttc tctttccgtc 901 tcccaattcg tattccgctc ctctcatctg aacacaatgt gcaagtttat ttatcttctc 961 gctttcattt cattaggacg tggggggaat tggtggaagg gggaaacaca caaaaggatg 1021 atggaaatga aataaggaca cacaatatgc aacaacattc aattcagaaa tatggaggaa 1081 ggtttaaaag aaaacataaa aatatataga ggaggaagga aaactagtaa aaaataagca 1141 aagaaattag gcgaacgatg agaattgtcc tcgcttggaa gttcctattc tctagaaagt 1201 ataggaactt ctaactaatc tgatttaaat tttcagggta gcggtggcag tggaggtacc 1261 ggcggaagcg gtatgggaag agcacccagc tttcttgtac aaagttggca ttataagaaa 1321 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat 1381 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg 1441 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaataatatc 1501 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca 1561 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg 1621 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg 1681 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt 1741 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa 1801 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac 1861 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc 1921 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg 1981 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga 2041 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact 2101 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat 2161 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg 2221 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa 2281 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt 2341 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat 2401 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc 2461 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt 2521 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 2581 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 2641 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt 2701 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 2761 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 2821 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 2881 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 2941 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 3001 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 3061 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 3121 acggttcctg gccttttgct ggccttttgc tcacatgtt //