LOCUS Exported 4813 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of ompR under PLtetO-1, output promoter PompF146. ACCESSION . VERSION . KEYWORDS pSR62 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4813) TITLE Rewired bacterial two-component systems JOURNAL Unpublished REFERENCE 2 (bases 1 to 4813) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4813 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 56..74 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 95..168 /label=PLtetO-1 promoter /note="modified phage lambda PL promoter with tet operator sites (Lutz and Bujard, 1997)" protein_bind 95..113 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 120..138 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" terminator 944..1015 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 1031..1058 /label=T7Te terminator /note="phage T7 early transcription terminator" terminator 1200..1243 /label=bacterial terminator /note="putative bacterial transcription terminator" CDS 1392..2105 /codon_start=1 /product="GFP variant that folds robustly even when fused to poorly folded proteins (Pedelacq et al., 2006)" /label=superfolder GFP /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" terminator 2132..2203 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 2219..2246 /label=T7Te terminator /note="phage T7 early transcription terminator" CDS 2367..2993 /codon_start=1 /gene="tetR" /product="tetracycline repressor TetR" /label=TetR /note="TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline." /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS" primer_bind complement(2443..2462) /label=Tet-R /note="Tetracycline resistance gene, reverse primer" terminator 2999..3085 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" rep_origin complement(3250..3838) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3330..3349) /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 3926..4020 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(4041..4700) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 4615..4634 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" ORIGIN 1 ttcgccagat atcgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag 61 gcgtatcacg aggccctttc gtcttcacct cgagtcccta tcagtgatag agattgacat 121 ccctatcagt gatagagata ctgagcacat cagcaggacg cactgaccgc ataaaggact 181 tagcaaagcc caattttaaa caaatgcaag agaactacaa gattctggtg gtcgatgacg 241 acatgcgcct gcgtgcgctg ctggaacgtt atctcaccga acaaggcttc caggttcgaa 301 gcgtcgctaa tgcagaacag atggatcgcc tgctgactcg tgaatctttc catcttatgg 361 tactggattt aatgttacct ggtgaagatg gcttgtcgat ttgccgacgt cttcgtagtc 421 agagcaaccc gatgccgatc attatggtga cggcgaaagg ggaagaagtg gaccgtatcg 481 taggcctgga gattggcgct gacgactaca ttccaaaacc gtttaacccg cgtgaactgc 541 tggcccgtat ccgtgcggtg ctgcgtcgtc aggcgaacga actgccaggc gcaccgtcac 601 aggaagaggc ggtaattgct ttcggtaagt tcaaacttaa cctcggtacg cgcgaaatgt 661 tccgcgaaga cgagccgatg ccgctcacca gcggtgagtt tgcggtactg aaggcactgg 721 tcagccatcc gcgtgagccg ctctcccgcg ataagctgat gaaccttgcc cgtggtcgtg 781 aatattccgc aatggaacgc tccatcgacg tgcagatttc gcgtctgcgc cgcatggtgg 841 aagaagatcc agcgcatccg cgttacattc agaccgtctg gggtctgggc tacgtctttg 901 taccggacgg ctctaaagca tgataataat ctagaccagg catcaaataa aacgaaaggc 961 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta 1021 gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tatagtaaat cactgcataa 1081 ttcgtgtcgc tcaaggcgca ctcccgttct ggataatgtt ttttgcgccg acatcataac 1141 ggttctggca aatattctga aatgagctgg ttagctagtc aggcgtatca cgaggcagaa 1201 tttcagataa aaaaaatcct tagctttcgc taaggatgat ttctggttcc ttaaatttta 1261 cttttggtta catatttttt ctttttgaaa ccaaatcttt atctttgtag cactttcacg 1321 gtagcgaaac gttagtttga atggaaagat gcctgcaggt gtcataaaaa aaaccatgag 1381 ggtaataaat aatgcgtaaa ggcgaagagc tgttcactgg tgtcgtccct attctggtgg 1441 aactggatgg tgatgtcaac ggtcataagt tttccgtgcg tggcgagggt gaaggtgacg 1501 caactaatgg taaactgacg ctgaagttca tctgtactac tggtaaactg ccggtacctt 1561 ggccgactct ggtaacgacg ctgacttatg gtgttcagtg ctttgctcgt tatccggacc 1621 atatgaagca gcatgacttc ttcaagtccg ccatgccgga aggctatgtg caggaacgca 1681 cgatttcctt taaggatgac ggcacgtaca aaacgcgtgc ggaagtgaaa tttgaaggcg 1741 ataccctggt aaaccgcatt gagctgaaag gcattgactt taaagaagac ggcaatatcc 1801 tgggccataa gctggaatac aattttaaca gccacaatgt ttacatcacc gccgataaac 1861 aaaaaaatgg cattaaagcg aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc 1921 agctggctga tcactaccag caaaacactc caatcggtga tggtcctgtt ctgctgccag 1981 acaatcacta tctgagcacg caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc 2041 atatggttct gctggagttc gtaaccgcag cgggcatcac gcatggtatg gatgaactgt 2101 acaaatgatg ataataatct agaccaggca tcaaataaaa cgaaaggctc agtcgaaaga 2161 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg 2221 ctcaccttcg ggtgggcctt tctgcgttta taggtacccg tggatcctct agagtcgacc 2281 tgcaggcatg caagcttttt acggctagct cagtcctagg tatagtgcta gcccagccag 2341 agaaacactc tttaacaggg ggtagtatga tgtctcgttt agataaaagt aaagtgatta 2401 acagcgcatt agagctgctt aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg 2461 cccagaagct aggtgtagag cagcctacat tgtattggca tgtaaaaaat aagcgggctt 2521 tgctcgacgc cttagccatt gagatgttag ataggcacca tactcacttt tgccctttag 2581 aaggggaaag ctggcaagat tttttacgta ataacgctaa aagttttaga tgtgctttac 2641 taagtcatcg cgatggagca aaagtacatt taggtacacg gcctacagaa aaacagtatg 2701 aaactctcga aaatcaatta gcctttttat gccaacaagg tttttcacta gagaatgcat 2761 tatatgcact cagcgcagtg gggcatttta ctttaggttg cgtattggaa gatcaagagc 2821 atcaagtcgc taaagaagaa agggaaacac ctactactga tagtatgccg ccattattac 2881 gacaagctat cgaattattt gatcaccaag gtgcagagcc agccttctta ttcggccttg 2941 aattgatcat atgcggatta gaaaaacaac ttaaatgtga aagtgggtct taaggcatca 3001 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt 3061 gaacgctctc ctgagtagga caaatccgcc gccctagacc tagggcgttc ggctgcggcg 3121 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 3181 atgcaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 3241 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 3301 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 3361 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 3421 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 3481 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 3541 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 3601 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 3661 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 3721 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 3781 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 3841 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 3901 gattttggtc atgactagtg cttggattct caccaataaa aaacgcccgg cggcaaccga 3961 gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc 4021 caagcgagct cgatatcaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat 4081 tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc 4141 agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg 4201 aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg 4261 gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag gttttcacca 4321 taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca 4381 ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca 4441 ctatcccata tcaccagctc accgtctttc attgccatac gaaattccgg atgagcattc 4501 atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg 4561 gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact 4621 gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat 4681 ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa 4741 aatacgaccg gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgggccgg 4801 ccaacgtctc att //