LOCUS Exported 3247 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Multisite gateway vector for 5' tagging with mKate2. ACCESSION . VERSION . KEYWORDS pME-mKate2-NS SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3247) TITLE Parton Lab Plasmids for in vivo cell biology in the zebrafish REFERENCE 2 (bases 1 to 3247) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3247 /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..802 /codon_start=1 /product="monomeric far-red fluorescent protein (Shcherbo et al., 2009)" /label=mKate2 /note="mammalian codon-optimized" /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY CDLPSKLGHR" protein_bind complement(804..903) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(920..939) /label=T7 /note="T7 promoter, forward primer" promoter complement(921..939) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(944..960) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(944..960) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1062..1081) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1073..1882 /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(1140..1159) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2029..2617 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2518..2537 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2771..2788 /label=L4440 /note="L4440 vector, forward primer" terminator 2947..2974 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 3066..3152 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 3201..3223 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 3215..3232 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 3216..3232 /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 tgagcgagct 121 gattaaggag aacatgcaca tgaagctgta catggagggc accgtgaaca accaccactt 181 caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca tgagaatcaa 241 ggcggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacca gcttcatgta 301 cggcagcaaa accttcatca accacaccca gggcatcccc gacttcttta agcagtcctt 361 ccccgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg tgctgaccgc 421 tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga tcagaggggt 481 gaacttccca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg aggcctccac 541 cgagaccctg taccccgctg acggcggcct ggaaggcaga gccgacatgg ccctgaagct 601 cgtgggcggg ggccacctga tctgcaactt gaagaccaca tacagatcca agaaacccgc 661 taagaacctc aagatgcccg gcgtctacta tgtggacaga agactggaaa gaatcaagga 721 ggccgacaaa gagacctacg tcgagcagca cgaggtggct gtggccagat actgcgacct 781 ccctagcaaa ctggggcaca gagacccagc tttcttgtac aaagttggca ttataagaaa 841 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat 901 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg 961 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc 1021 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca 1081 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg 1141 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg 1201 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt 1261 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa 1321 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac 1381 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc 1441 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg 1501 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga 1561 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact 1621 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat 1681 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg 1741 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa 1801 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt 1861 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat 1921 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc 1981 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt 2041 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 2101 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 2161 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt 2221 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 2281 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 2341 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 2401 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 2461 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 2521 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 2581 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 2641 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga 2701 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac 2761 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc 2821 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa 2881 agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga 2941 aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat 3001 ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc 3061 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc 3121 cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt 3181 aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct 3241 cgggccc //