LOCUS Exported 4055 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION BIOFAB reporter plasmid for measuring tetAC termination efficiency. ACCESSION . VERSION . KEYWORDS pFAB820 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4055) 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 4055) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4055 /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 1364..1375 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 1382..2095 /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(1412..1440) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 2035..2055 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" terminator 2131..2202 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 2218..2245 /label=T7Te terminator /note="phage T7 early transcription terminator" primer_bind complement(2316..2333) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2450..2995) /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(2530..2549) /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 3109..3203 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(3234..4028) /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(3345..3364) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 3955..3974 /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 gtgattattt cgtcaccaaa tgatgttatt 1201 ccgcgaaata taatgaccct cttgataacc caagagggca ttttttacga taaagaagat 1261 ttagcttcaa ataaaacggc ggactcacta tagagggaca aactcaaggt cattcgcaag 1321 agtggccttt atgattgacc ttcttccggt taatcaattc attaaagagg agaaaggtac 1381 catgagcaaa ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg 1441 tgatgttaat gggcacaaat tttctgtccg tggagagggt gaaggtgatg ctacaaacgg 1501 aaaactcacc cttaaattta tttgcactac tggaaaacta cctgttccgt ggccaacact 1561 tgtcactact ctgacctatg gtgttcaatg cttttcccgt tatccggatc acatgaaacg 1621 gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaacgca ctatatcttt 1681 caaagatgac gggacctaca agacgcgtgc tgaagtcaag tttgaaggtg atacccttgt 1741 taatcgtatc gagttaaagg gtattgattt taaagaagat ggaaacattc ttggacacaa 1801 actcgagtac aactttaact cacacaatgt atacatcacg gcagacaaac aaaagaatgg 1861 aatcaaagct aacttcaaaa ttcgccacaa cgttgaagat ggttccgttc aactagcaga 1921 ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta 1981 cctgtcgaca caatctgtcc tttcgaaaga tcccaacgaa aagcgtgacc acatggtcct 2041 tcttgagttt gtaactgctg ctgggattac acatggcatg gatgagctct acaaataagg 2101 atcggttgtc gagtaaggat ctccaggcat caaataaaac gaaaggctca gtcgaaagac 2161 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag tcacactggc 2221 tcaccttcgg gtgggccttt ctgcgtttat aggatcctaa ctcgagctga agccaatagg 2281 gataacaggg taatcctagg gatatattcc gcttcctcgc tcactgactc gctacgctcg 2341 gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg gcggagattt cctggaagat 2401 gccaggaaga tacttaacag ggaagtgaga gggccgcggc aaagccgttt ttccataggc 2461 tccgcccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga 2521 caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc 2581 ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct cattccacgc 2641 ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca cgaacccccc 2701 gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggaaaga 2761 catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt tagtcttgaa 2821 gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg ctcctccaag 2881 ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa 2941 ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga 3001 tcatcttatt aatcagataa aatatttcta gatttcagtg caatttatct cttcaaatgt 3061 agcacctgaa gtcagcccca tacgatataa gttgttacta gtgcttggat tctcaccaat 3121 aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat 3181 tactggatct atcaacagga gtccaagcga gctctcgaac cccagagtcc cgctcagaag 3241 aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa 3301 agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc 3361 aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa 3421 aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga 3481 tcctcgccgt cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc 3541 tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct 3601 cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc 3661 agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac 3721 aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca 3781 acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc 3841 tcgtcctgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc 3901 ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag 3961 tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt 4021 tcaatcatgc gaaacgatcc tcatcctgtc tcttg //