LOCUS Exported 5699 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION SapTrap Halotag + Cbr-unc-119 donor plasmid. ACCESSION . VERSION . KEYWORDS pMLS254 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5699) 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 5699) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5699 /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 complement(1217..1250) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS 3214..3225 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" protein_bind complement(3374..3407) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind complement(3824..3923) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(3940..3959) /label=T7 /note="T7 promoter, forward primer" promoter complement(3941..3959) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(3964..3980) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3964..3980) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4082..4101) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 4093..4902 /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(4160..4179) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 5049..5637 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5538..5557 /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 tcttcgatga tggctgaaat tggtactggc tttccttttg atccccacta 721 cgttgaggtt cttggagagc gcatgcacta cgttgatgtt ggaccacgcg atggaacccc 781 agtccttttc cttcacggaa acccaacttc ttcttacgtt tggcgcaaca tcatcccaca 841 tgttgctcca acccaccgct gcatcgctcc agatcttatc ggaatgggaa agtctgataa 901 gccagatctt ggatacttct tcgatgatca cgtccgtttc atggatgctt tcatcgaggc 961 ccttggactt gaggaggttg tccttgttat ccacgattgg ggatccgctc ttggattcca 1021 ctgggctaag cgcaacccag agcgcgttaa gggaatcgct ttcatggaat tcatccgccc 1081 aatcccaact tgggatgaat ggccagagtt cgctcgcgag accttccaag ctttccgcac 1141 caccgatgtt ggacgtaagc ttatcatcga tcaaaacgtt ttcatcgaag gtaagtttaa 1201 acatgatttt actaacataa cttcgtataa tgtatgctat acgaagttat ttctagacat 1261 tctctaatga aaaaatcttt cagttgaaat tgaaaatgag ttaaagttgg agtttttatt 1321 gaaaacagat ttccgtgtga ttagtgtttt tagcgagtgt gacaggacag cgaaaaaata 1381 tagaaacaag gggggaactg aaaagcttag gaatgcattg aacatgagaa ggggaagggg 1441 aaggaacaaa ctagacagga attattggaa tttaatcaca tttggagttt tttttctatt 1501 cgacagaata attatccaga acatttttgt attaaatatt tatgcatcat atgagtagtc 1561 ggctttgttg tgcatgacga gtttgttatc gacgaaatag aagctgtcag aacgagtctc 1621 gtttggattg ttgatcatgt cgtccactga aaaagagatt agtcttttga attgtacttt 1681 ttagaataat gactcactga gttgttgaga gagttgaggg aactcataga tatgttcaca 1741 gttgtttcgt gaattcggaa tacagaatcc gaattcaaag tcaaaacact tcagaaggcg 1801 atctttgaag aagtgacgtt cgatcattcg gaaatgatgg attgggatgt ctcctacttt 1861 gaattccaca gttgctccga ccgttttgag tttcaaaaag tttggagcaa atctataacg 1921 cacgtatctt gccgactcct gtggcgactc atcattctcc tgatcgttct ccggtttggc 1981 gatctcgaaa agcacttgct cagtgtccag atcacggatt tggaacttgg tgaactcgat 2041 gttatagatg ttcgcagatg gggagcataa gaatcctaaa tttatgtttt aaactgaaat 2101 ccaaagggag caagatacct tgagtgattc ccggaagtgc taaaacgtcg ttcggagtga 2161 tttgagcttt cttcgcaagc tccgattccg ttgtgattcc ttgttcggtg cttggtggtg 2221 gccgtggcat ctggaaatat ggaaaagttc aacaaaaaga aaagagaaaa gaatgaaatc 2281 ggatatcaag agttagttga gcggtttctc tagttttctg agtctcacct gcgacgggaa 2341 ggtcgccgag ccgggtggaa tcgattgttg ttgctcggct ttcatatcgg tttggttgga 2401 agcggctgaa aacggaaaga agtggaagaa ggaaaagagt gtggtgtgac aggaaaatgg 2461 taattagagg gtgccaaata accagctata ttttgttttt ttttgaaaac atttttaaaa 2521 agaaaaatac gataatgata tcagatggat ttccggaaaa ctggtatgaa aaatttcaac 2581 ctttttgagt acatgtaatc aaaatacact ttgtaaatta tcatttttat tgaaactcca 2641 ccatttttct atttataacg ctaataattt gaaaaagaaa cctgttgcga accgcggggt 2701 gaatcccaaa aacgaatgcg ttttggtgga gtgattgatt cgaatcgaag aagaaaaaga 2761 agaagacgtg gaatagagag ctcactctta accgagcagc acacaccgac agaaaaaaaa 2821 atgaaatgaa tgagggtctt cttcttcttc ttcttcgaat gattgacaga aatgggaaaa 2881 agaggaagat tgagaaggga aaaaggaagg agaaaagaag cagaagaaga cgtcagagag 2941 gagaggaacg agcggaaaag cagcgggcgc aagtcataga agtagcagag ctggggagaa 3001 gaagacacta tccaagaaag gaatgacgag agagtatgca aaggggtata gggtgcagac 3061 agaataggaa cagaataaca gatgatgagc caagaagagt tgaaaagggc gatgaatttg 3121 tcatgtaact taatttgggt caatttgagc atgatgaatt gaaatcatcc cttgttggga 3181 gttaataacc ggtttgttat cagaaaccct gtaatagaag ggcgccctaa ctttgagcca 3241 attcatcccg gtttctgtca aatatatcaa aaagtggtca actgacaaat tgtttttgat 3301 attataataa acattttatc cgttaacaat tttcgaatac tttttacaag gacttggata 3361 aattggctca aagataactt cgtataatgt atgctatacg aagttattaa ctaatctgat 3421 ttaaattttc aggaaccctt ccaatgggag ttgttcgtcc acttaccgag gttgagatgg 3481 atcactaccg cgagccattc cttaacccag ttgatcgcga gccactttgg cgcttcccaa 3541 acgaacttcc aatcgctgga gagccagcta acatcgttgc tcttgttgag gagtacatgg 3601 attggcttca ccaatctcca gttccaaagc ttcttttctg gggaactcca ggagttctta 3661 tcccaccagc tgaggctgcc cgtcttgcta agtctcttcc aaactgcaag gctgtcgata 3721 tcggaccagg acttaacctt cttcaagagg ataacccaga tcttatcgga tctgagatcg 3781 ctcgttggct ttctaccctt gagatctccg gaggtcgaag agcacccagc tttcttgtac 3841 aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc 3901 agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac 3961 atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt 4021 gcacaagata aaataatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat 4081 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac 4141 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg 4201 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa 4261 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt 4321 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc 4381 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa 4441 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat 4501 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac 4561 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc 4621 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat 4681 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga 4741 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag 4801 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg 4861 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg 4921 ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa 4981 tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa 5041 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 5101 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 5161 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 5221 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 5281 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 5341 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 5401 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 5461 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 5521 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 5581 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 5641 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtt //