LOCUS Exported 3282 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Membrane targeted KOFP2 (orange) fluorophore for use with 5' promoters.. ACCESSION . VERSION . KEYWORDS pME-mKOFP2-CAAX SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3282) AUTHORS Don EK, Formella I, Badrock AP, Hall TE, Morsch M, Hortle E, Hogan A, Chow S, Gwee SS, Stoddart JJ, Nicholson G, Chung R, Cole NJ TITLE A Tol2 Gateway-Compatible Toolbox for the Study of the Nervous System and Neurodegenerative Disease. JOURNAL Zebrafish. 2016 Sep 15. PUBMED 27631880 REFERENCE 2 (bases 1 to 3282) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3282 /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" regulatory 678..687 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 684..1337 /codon_start=1 /product="monomeric Kusabira-Orange 2 fluorescent protein" /label=mKO2 /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS" protein_bind complement(1407..1506) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1523..1542) /label=T7 /note="T7 promoter, forward primer" promoter complement(1524..1542) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1547..1563) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1547..1563) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1665..1684) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1676..2485 /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(1743..1762) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2632..3220 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3121..3140 /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 agcaggctga attcgccgcc accatggtga gtgtgattaa acctgagatg aagatgcgct 721 actacatgga cggatctgtg aacggccacg agtttaccat cgagggcgag ggaaccggca 781 gaccttacga gggacaccag gaaatgaccc tgagagtcac catggccgag ggcggaccta 841 tgcctttcgc cttcgacctg gtgtctcacg tgttctgcta cggacacaga gtgttcacca 901 agtaccctga ggaaatcccc gactacttca agcaggcctt ccctgaggga ctgagctggg 961 agagaagcct ggaatttgag gacggcggaa gcgctagcgt gtccgcccac atcagcctga 1021 gaggaaacac cttctaccac aagagcaagt tcacaggcgt gaacttccct gccgacggac 1081 ctatcatgca gaaccagagc gtggactggg agcctagcac cgagaagatc accgccagcg 1141 acggcgtgct gaagggcgac gtgaccatgt acctgaagct ggaaggcgga ggcaaccaca 1201 agtgccagat gaagaccacc tacaaggccg ccaaagaaat cctggaaatg ccaggcgacc 1261 actacatcgg acacagactg gtccgcaaga ccgagggaaa catcaccgag caggtcgagg 1321 acgccgtggc tcacagcaag ctgaaccctc cagacgagag cggacctgga tgcatgagct 1381 gcaaatgcgt gctctcctga actagtaccc agctttcttg tacaaagttg gcattataag 1441 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat 1501 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc 1561 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat 1621 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag 1681 ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata 1741 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta 1801 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga 1861 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat 1921 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa 1981 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct 2041 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga 2101 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag 2161 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa 2221 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct 2281 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga 2341 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca 2401 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca 2461 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag 2521 cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta 2581 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc 2641 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt 2701 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc 2761 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc 2821 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg 2881 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg 2941 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga 3001 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc 3061 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg 3121 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg 3181 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt 3241 tttacggttc ctggcctttt gctggccttt tgctcacatg tt //