LOCUS Exported 3527 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Encodes the fluorescent protein Rosmarinus for constitutive expression in E. coli.. ACCESSION . VERSION . KEYWORDS pRSM32 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3527) AUTHORS Molina RS, Tran TM, Campbell RE, Lambert GG, Salih A, Shaner NC, Hughes TE, Drobizhev M TITLE Blue-Shifted Green Fluorescent Protein Homologues Are Brighter than Enhanced Green Fluorescent Protein under Two-Photon Excitation. JOURNAL J Phys Chem Lett. 2017 Jun 15;8(12):2548-2554. doi: 10.1021/acs.jpclett.7b00960. Epub 2017 May 25. PUBMED 28530831 REFERENCE 2 (bases 1 to 3527) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3527 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 104..125 /label=F1ori-F /note="F1 origin, forward primer" promoter 288..392 /gene="bla" /label=AmpR promoter CDS 393..1187 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(447..466) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 1057..1076 /label=Neo-F /note="Neomycin resistance gene, forward primer" rep_origin 1358..1946 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1847..1866 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2100..2117 /label=L4440 /note="L4440 vector, forward primer" primer_bind 2202..2221 /label=T7 /note="T7 promoter, forward primer" promoter 2202..2220 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 2304..2326 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 2346..2363 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" CDS 2367..2399 /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /label=T7 tag (gene 10 leader) /note="promotes efficient translation in E. coli" /translation="MASMTGGQQMG" CDS 2403..2426 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /label=Xpress(TM) tag /translation="DLYDDDDK" primer_bind complement(3176..3194) /label=T7 Term /note="T7 terminator, reverse primer" terminator 3190..3237 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 3334..262 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3421..3440) /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag 61 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa 121 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg 181 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac 241 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta 301 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat 361 aaatgcttca ataatattga aaaaggaaga gtatgattga acaagatgga ttgcacgcag 421 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg 481 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca 541 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc 601 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg 661 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg 721 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta 781 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag 841 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac 901 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg 961 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg 1021 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg 1081 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg 1141 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgactg tcagaccaag 1201 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg 1261 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact 1321 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg 1381 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 1441 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 1501 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 1561 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 1621 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 1681 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 1741 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 1801 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 1861 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 1921 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg 1981 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata 2041 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca 2101 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc 2161 gttggccgat tcattaatgc aggatctcga tcccgcgaaa ttaatacgac tcactatagg 2221 gagaccacaa cggtttccct ctagaatcac cgagtttatt cttgacacct gatgcgatga 2281 atgatataat aggaaagtac tgttttgttt aactttaaga aggagatata catatgcggg 2341 gttctcatca tcatcatcat catggtatgg ctagcatgac tggtggacag caaatgggtc 2401 gggatctgta cgacgatgac gataaggatc cgtcagtgat aaagagcgtc atgaaaatta 2461 agctgcatat ggacggcatc gtgaacggtc acaaattcat gatcaccggt gagggggaag 2521 gcaagccatt tgaggggact cataccatta tcctgaaggt gaaggaagga ggaccactgc 2581 ctttcgcata tgatatcctc actacagcct ttcagtacgg caatcgggtc ttcacaaaat 2641 atcccaagga tattcctgac tacttcaaac agtcattccc ggagggctac agttgggagc 2701 ggtccatgac ctttgaggat cagggcgtgt gtaccgtgac ctctgacatc aaactggaag 2761 gagactattt cacctacgat atcaggtttc acggagtcaa tttccccgcc ggcggtcccg 2821 tcatgcagaa aaagactctg cggtgggaac caagcaccga gaacatgtac gtgcgagacg 2881 gcgtgctggt aggcgaggtg gagagaactc tgctgctgga gggcaataaa catcaccgct 2941 gtaacttcag aacaacttac aaagcaaaga aagaagtagt cctgccagaa taccactttg 3001 ttgaccaccg gatagagatc ttggggcatg acaaggacta taacaatgtg gttgtgtatg 3061 aaaacgcagt ggctcggcaa caggcctcta ccctgccaag caaggcgaag taggaattcg 3121 aagcttgatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct 3181 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg 3241 aaaggaggaa ctatatccgg atctggcgta atagcgaaga ggccagcacc gatcgccctt 3301 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg 3361 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg 3421 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc 3481 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcac //