LOCUS Exported 4036 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION BIOFAB reporter plasmid for measuring rpoC termination efficiency. ACCESSION . VERSION . KEYWORDS pFAB1910 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4036) AUTHORS Mutalik VK, Guimaraes JC, Cambray G, Lam C, Christoffersen MJ, Mai QA, Tran AB, Paull M, Keasling JD, Arkin AP, Endy D TITLE Precise and reliable gene expression via standard transcription and translation initiation elements. JOURNAL Nat Methods. 2013 Apr;10(4):354-60. doi: 10.1038/nmeth.2404. Epub 2013 Mar 10. PUBMED 23474465 REFERENCE 2 (bases 1 to 4036) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4036 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 320..393 /label=PLtetO-1 promoter /note="modified phage lambda PL promoter with tet operator sites (Lutz and Bujard, 1997)" protein_bind 320..338 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 345..363 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" RBS 407..418 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 425..1102 /codon_start=1 /product="monomeric derivative of DsRed (Campbell et al., 2002)" /label=mRFP1 /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS TGA" RBS 1345..1356 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 1363..2076 /codon_start=1 /product="GFP variant that folds robustly even when fused to poorly folded proteins (Pedelacq et al., 2006)" /label=superfolder GFP /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" primer_bind complement(1393..1421) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 2016..2036 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" terminator 2112..2183 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 2199..2226 /label=T7Te terminator /note="phage T7 early transcription terminator" primer_bind complement(2297..2314) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2431..2976) /direction=LEFT /label=p15A ori /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin. " primer_bind complement(2511..2530) /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 3090..3184 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(3215..4009) /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 GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3326..3345) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 3936..3955 /label=Neo-R /note="Neomycin resistance gene, reverse primer" ORIGIN 1 atcagatcat gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac 61 tttgcagggc ttcccaacct taccagaggg cgccccagct ggcaattccg acgtctaggg 121 ataacagggt aattacggcc ccagaattca aaagatctta agtaagtaag agtatacgta 181 tatcggctaa aacgtattaa ggcgcttcgg cgcctttttt tatgggggta ttttcatccc 241 aatccacacg tccaacgcac agcaaacacc acgtcgaccc tatcagctgc gtgctttcta 301 tgagtcgttg ctgcatatct ccctatcagt gatagagatt gacatcccta tcagtgatag 361 agatactgag cacatcagca ggacgcactg acctttggaa ttcattaaag aggagaaagg 421 taccatggcg agtagcgaag acgttatcaa agagttcatg cgtttcaaag ttcgtatgga 481 aggttccgtt aacggtcacg agttcgaaat cgaaggtgaa ggtgaaggtc gtccgtacga 541 aggtacccag accgctaaac tgaaagttac caaaggtggt ccgctgccgt tcgcttggga 601 catcctgtcc ccgcagttcc agtacggttc caaagcttac gttaaacacc cggctgacat 661 cccggactac ctgaaactgt ccttcccgga aggtttcaaa tgggaacgtg ttatgaactt 721 cgaagacggt ggtgttgtta ccgttaccca ggactcctcc ctgcaagacg gtgagttcat 781 ctacaaagtt aaactgcgtg gtaccaactt cccgtccgac ggtccggtta tgcagaaaaa 841 aaccatgggt tgggaagctt ccaccgaacg tatgtacccg gaagacggtg ctctgaaagg 901 tgaaatcaaa atgcgtctga aactgaaaga cggtggtcac tacgacgctg aagttaaaac 961 cacctacatg gctaaaaaac cggttcagct gccgggtgct tacaaaaccg acatcaaact 1021 ggacatcacc tcccacaacg aagactacac catcgttgaa cagtacgaac gtgctgaagg 1081 tcgtcactcc accggtgctt aataatctgc tgagtgatag actcaaggtc gctcctagcg 1141 agtggccttt atgattatca ctttacttat gtgatgtaat cgttaatccg caaataacgt 1201 aaaaacccgc ttcggcgggt ttttttatgg ggggagttta gggaaagagc atttgtcagg 1261 cggactcact atagagggac aaactcaagg tcattcgcaa gagtggcctt tatgattgac 1321 cttcttccgg ttaatcaatt cattaaagag gagaaaggta ccatgagcaa aggagaagaa 1381 cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa 1441 ttttctgtcc gtggagaggg tgaaggtgat gctacaaacg gaaaactcac ccttaaattt 1501 atttgcacta ctggaaaact acctgttccg tggccaacac ttgtcactac tctgacctat 1561 ggtgttcaat gcttttcccg ttatccggat cacatgaaac ggcatgactt tttcaagagt 1621 gccatgcccg aaggttatgt acaggaacgc actatatctt tcaaagatga cgggacctac 1681 aagacgcgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatcgtat cgagttaaag 1741 ggtattgatt ttaaagaaga tggaaacatt cttggacaca aactcgagta caactttaac 1801 tcacacaatg tatacatcac ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa 1861 attcgccaca acgttgaaga tggttccgtt caactagcag accattatca acaaaatact 1921 ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtcgac acaatctgtc 1981 ctttcgaaag atcccaacga aaagcgtgac cacatggtcc ttcttgagtt tgtaactgct 2041 gctgggatta cacatggcat ggatgagctc tacaaataag gatcggttgt cgagtaagga 2101 tctccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct 2161 gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg ggtgggcctt 2221 tctgcgttta taggatccta actcgagctg aagccaatag ggataacagg gtaatcctag 2281 ggatatattc cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag 2341 cggaaatggc ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca 2401 gggaagtgag agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca 2461 tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca 2521 ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg 2581 tgtcattccg ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt 2641 aggcagttcg ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg 2701 ccttatccgg taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg 2761 cagcagccac tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc 2821 taaactgaaa ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa 2881 gagttggtag ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca 2941 gagcaagaga ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata 3001 aaatatttct agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc 3061 atacgatata agttgttact agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa 3121 ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg 3181 agtccaagcg agctctcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat 3241 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag 3301 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc 3361 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca 3421 tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc 3481 gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat 3541 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg 3601 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag 3661 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca 3721 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc 3781 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca 3841 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga 3901 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct 3961 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc 4021 ctcatcctgt ctcttg //