LOCUS Exported 4044 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION BIOFAB reporter plasmid for measuring RNAI termination efficiency. ACCESSION . VERSION . KEYWORDS pFAB807 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4044) 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 4044) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4044 /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 1353..1364 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 1371..2084 /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(1401..1429) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 2024..2044 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" terminator 2120..2191 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 2207..2234 /label=T7Te terminator /note="phage T7 early transcription terminator" primer_bind complement(2305..2322) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2439..2984) /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(2519..2538) /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 3098..3192 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(3223..4017) /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(3334..3353) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 3944..3963 /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 gtgatgagtt ggtagctctt gatccggcaa 1201 acaaaccacc gttggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa 1261 aaaaggggcg gactcactat agagggacaa actcaaggtc attcgcaaga gtggccttta 1321 tgattgacct tcttccggtt aatcaattca ttaaagagga gaaaggtacc atgagcaaag 1381 gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt gatgttaatg 1441 ggcacaaatt ttctgtccgt ggagagggtg aaggtgatgc tacaaacgga aaactcaccc 1501 ttaaatttat ttgcactact ggaaaactac ctgttccgtg gccaacactt gtcactactc 1561 tgacctatgg tgttcaatgc ttttcccgtt atccggatca catgaaacgg catgactttt 1621 tcaagagtgc catgcccgaa ggttatgtac aggaacgcac tatatctttc aaagatgacg 1681 ggacctacaa gacgcgtgct gaagtcaagt ttgaaggtga tacccttgtt aatcgtatcg 1741 agttaaaggg tattgatttt aaagaagatg gaaacattct tggacacaaa ctcgagtaca 1801 actttaactc acacaatgta tacatcacgg cagacaaaca aaagaatgga atcaaagcta 1861 acttcaaaat tcgccacaac gttgaagatg gttccgttca actagcagac cattatcaac 1921 aaaatactcc aattggcgat ggccctgtcc ttttaccaga caaccattac ctgtcgacac 1981 aatctgtcct ttcgaaagat cccaacgaaa agcgtgacca catggtcctt cttgagtttg 2041 taactgctgc tgggattaca catggcatgg atgagctcta caaataagga tcggttgtcg 2101 agtaaggatc tccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg 2161 ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg 2221 tgggcctttc tgcgtttata ggatcctaac tcgagctgaa gccaataggg ataacagggt 2281 aatcctaggg atatattccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact 2341 gcggcgagcg gaaatggctt acgaacgggg cggagatttc ctggaagatg ccaggaagat 2401 acttaacagg gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct 2461 gacaagcatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa 2521 agataccagg cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg 2581 tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag 2641 ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg ttcagtccga 2701 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac atgcaaaagc 2761 accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag tcatgcgccg 2821 gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc cagttacctc 2881 ggttcaaaga gttggtagct cagagaacct tcgaaaaacc gccctgcaag gcggtttttt 2941 cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat catcttatta 3001 atcagataaa atatttctag atttcagtgc aatttatctc ttcaaatgta gcacctgaag 3061 tcagccccat acgatataag ttgttactag tgcttggatt ctcaccaata aaaaacgccc 3121 ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta 3181 tcaacaggag tccaagcgag ctctcgaacc ccagagtccc gctcagaaga actcgtcaag 3241 aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa 3301 gcggtcagcc cattcgccgc caagctcttc agcaatatca cgggtagcca acgctatgtc 3361 ctgatagcgg tccgccacac ccagccggcc acagtcgatg aatccagaaa agcggccatt 3421 ttccaccatg atattcggca agcaggcatc gccatgggtc acgacgagat cctcgccgtc 3481 gggcatgcgc gccttgagcc tggcgaacag ttcggctggc gcgagcccct gatgctcttc 3541 gtccagatca tcctgatcga caagaccggc ttccatccga gtacgtgctc gctcgatgcg 3601 atgtttcgct tggtggtcga atgggcaggt agccggatca agcgtatgca gccgccgcat 3661 tgcatcagcc atgatggata ctttctcggc aggagcaagg tgagatgaca ggagatcctg 3721 ccccggcact tcgcccaata gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac 3781 agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag 3841 ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga 3901 cagccggaac acggcggcat cagagcagcc gattgtctgt tgtgcccagt catagccgaa 3961 tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg 4021 aaacgatcct catcctgtct cttg //