LOCUS Exported 2884 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Multisite gateway vector for 3' tagging with the CaaX domain from Kras. ACCESSION . VERSION . KEYWORDS p3E-CaaX(tK) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2884) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 2884) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2884 /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 complement(624..748) /gene="mutant version of attR" /label=attR2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" polyA_signal 807..941 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(831..850) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 885..904 /label=SV40pA-R /note="SV40 polyA, reverse primer" protein_bind complement(993..1088) /gene="mutant version of attL" /label=attL3 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1125..1144) /label=T7 /note="T7 promoter, forward primer" promoter complement(1126..1144) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1149..1165) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1149..1165) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1267..1286) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1278..2087 /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(1345..1364) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2234..2822 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2723..2742 /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 tcgggccctg cagctctaga gctcgaattc tacaggtcac 601 taataccatc taagtagttg gttcatagtg actgcatatg ttgtgtttta cagtattatg 661 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt 721 ctcgttcaac tttcttgtac aaagtgggaa agaagaagaa gaagaagagc aagaccaagt 781 gcgtgatcat gtgatcgtat tacgtagatc cagacatgat aagatacatt gatgagtttg 841 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta 901 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc 961 attttatgtt tcaggttcag ggggaggtgt ggcaacttta ttatacaaag ttggcattat 1021 aaaaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat 1081 cattatttgg agctccatgg tagcgttaac gcggccgcga tatcccctat agtgagtcgt 1141 attacatggt catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt 1201 acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca 1261 gtaatacaag gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt 1321 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag 1381 gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg 1441 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg 1501 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac 1561 tcaccactgc gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag 1621 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt 1681 gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga 1741 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac 1801 aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg 1861 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg 1921 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg 1981 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg 2041 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta 2101 attggttgta acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac 2161 caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga 2221 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 2281 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 2341 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt 2401 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 2461 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 2521 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 2581 tggagcgaac gacctacacc gaactgagat acctacagcg tgagcattga gaaagcgcca 2641 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 2701 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 2761 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 2821 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca 2881 tgtt //