LOCUS Exported 2602 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Multisite gateway vector for 5' tagging with the actin binding motif Lifeact. ACCESSION . VERSION . KEYWORDS pME-Lifeact SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2602) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 2602) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2602 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 1..100 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" regulatory 101..110 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 107..157 /codon_start=1 /product="peptide that binds filamentous actin (Riedl et al., 2008)" /label=Lifeact /note="first 17 amino acids of budding yeast Abp140" /translation="MGVADLIKKFESISKEE" protein_bind complement(159..258) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(275..294) /label=T7 /note="T7 promoter, forward primer" promoter complement(276..294) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(299..315) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(299..315) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(417..436) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 428..1237 /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(495..514) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 1384..1972 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1873..1892 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2126..2143 /label=L4440 /note="L4440 vector, forward primer" terminator 2302..2329 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 2421..2507 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 2556..2578 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2570..2587 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2571..2587 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa 61 tgctttttta taatgccaac tttgtacaaa aaagcaggct gccaccatgg gcgtggccga 121 cctgatcaag aagttcgaga gcatcagcaa ggaggagaac ccagctttct tgtacaaagt 181 tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca 241 aaataaaatc attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt 301 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca 361 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag 421 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga 481 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat 541 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag 601 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc 661 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc 721 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat 781 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc 841 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt 901 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa 961 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc 1021 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt 1081 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc 1141 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa 1201 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta 1261 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct 1321 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc 1381 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct 1441 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg 1501 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca 1561 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc 1621 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga 1681 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac 1741 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga 1801 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag 1861 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg 1921 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag 1981 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc 2041 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc 2101 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc 2161 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag 2221 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca 2281 ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga 2341 tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg 2401 ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag 2461 ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt 2521 ccctactctc gcgttaacgc tagcatggat gttttcccag tcacgacgtt gtaaaacgac 2581 ggccagtctt aagctcgggc cc //