LOCUS SEGA053 5534 bp ds-DNA linear UNA 17-JUN-2021 DEFINITION LMBP 12889. ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 5534) AUTHORS Morten Norholm Group TITLE Direct Submission JOURNAL Exported Jun 17, 2021 from SnapGene 4.2.11 COMMENT SEGA landing pad: PRha-low-YebF-sfGFP-T-tetA-S2P11-S3P22-rrnC FEATURES Location/Qualifiers source 1..5534 /organism="unspecified" /mol_type="genomic DNA" source 2656..2718 /organism="Escherichia coli str. K-12 substr. MG1655" /strain="K-12" /sub_strain="MG1655" /mol_type="genomic DNA" /db_xref="taxon:511145" CDS 1..528 /codon_start=3 /label=glmS /translation="VHGLQALPSRIEQMLSQDKRIEALAEDFSDKHHALFLGRGDQYPI ALEGALKLKEISYIHAEAYAAGELKHGPLALIDADMPVIVVAPNNELLEKLKSNIEEVR ARGGQLYVFADQDAGFVSSDNMHIIEMPHVEEVIAPIFYTVPLQLLAYHVALIKGTDVD QPRNLAKSVTVE" misc_feature 538..594 /label=L3S2P11 Terminator CDS complement(607..1452) /codon_start=1 /gene="E. coli rhaR" /product="transcriptional activator of rhaR and rhaS" /label=rhaR /note="needed for rhamnose-dependent activation of the rhaB promoter" /translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK D" CDS complement(1526..2362) /codon_start=1 /gene="E. coli rhaS" /product="positive regulator of the rhaB promoter" /label=rhaS /note="activated by L-rhamnose" /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ" promoter 2508..2626 /label=rhaB promoter /note="promoter of the E. coli rhaBAD operon, conferring tight induction with L-rhamnose and repression with D-glucose in the presence of RhaR and RhaS (Giacalone et al., 2006)" RBS 2638..2649 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" misc_feature 2650..2664 /label=TIR gene 2656..2718 /gene="yebF" /gene_synonym="ECK1848; JW1836" /locus_tag="b1847" /label=yebF /db_xref="EcoGene:EG11807" /db_xref="GeneID:946363" CDS 2656..2718 /codon_start=1 /transl_table=11 /gene="yebF" /gene_synonym="ECK1848; JW1836" /locus_tag="b1847" /product="extracellular Colicin M immunity family protein" /label=yebF /db_xref="GI:145698278" /db_xref="ASAP:ABE-0006141" /db_xref="UniProtKB/Swiss-Prot:P33219" /db_xref="EcoGene:EG11807" /db_xref="GeneID:946363" /protein_id="NP_416361.2" /translation="MKKRGAFLGLLLVSACASVFA" CDS 2719..3009 /codon_start=1 /label=YebF /translation="ANNETSKSVTFPKCEDLDAAGIAASVKRDYQQNRVARWADDQKIV GQADPVAWVSLQDIQGKDDKWSVPLTVRGKSADIHYQVSVDCKAGMAEYQRR" CDS 3010..3720 /codon_start=1 /label=sfGFP /translation="RKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGT YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF VTAAGITHGMDELYK" misc_feature 3724..3772 /label=L3S3P22 Terminator (synthetic) promoter 3783..3811 /label=Promoter 3BCD2 CDS 3851..5041 /codon_start=1 /gene="tet" /product="tetracycline efflux protein" /label=TcR /note="confers resistance to tetracycline" /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM QLVGQVPAALWVIFGEDRFRWSATMSGLSLAVFGILHALAQAFVTGPATKRFGEKQAII AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST" misc_feature 5022..5071 /label=HR3 terminator 5052..5092 /label=rrnC misc_feature 5093..5142 /label=homology CDS 5287..5534 /codon_start=1 /label=pstS /translation="MKVMRTTVATVVAATLSMSAFSVFAEASLTGAGATFPAPVYAKWA DTYQKETGNKVNYQGIGSSGGVKQIIANTVDFGASDA" ORIGIN 1 gtgcatggtc tgcaggcgct gccgagccgt attgagcaga tgctgtctca ggacaaacgc 61 attgaagcgc tggcagaaga tttctctgac aaacatcacg cgctgttcct gggccgtggc 121 gatcagtacc caatcgcgct ggaaggcgca ttgaagttga aagagatctc ttacattcac 181 gctgaagcct acgctgctgg cgaactgaaa cacggtccgc tggcgctaat tgatgccgat 241 atgccggtta ttgttgttgc accgaacaac gaattgctgg aaaaactgaa atccaacatt 301 gaagaagttc gcgcgcgtgg cggtcagttg tatgtcttcg ccgatcagga tgcgggtttt 361 gtaagtagcg ataacatgca catcatcgag atgccgcatg tggaagaggt gattgcaccg 421 atcttctaca ccgttccgct gcagctgctg gcttaccatg tcgcgctgat caaaggcacc 481 gacgttgacc agccgcgtaa cctggcaaaa tcggttacgg ttgagtaata aatggatctc 541 ggtaccaaat tccagaaaag agacgctttc gagcgtcttt tttcgttttg gtccgcgttg 601 cgcttaatct ttctgcgaat tgagatgacg ccactggctg ggcgtcatcc cggtttcccg 661 ggtaaacacc accgaaaaat agttactatc ttcaaagcca cattcggtcg aaatatcact 721 gattaacagg cggctatgct ggagaagata ttgcgcatga cacactctga cctgtcgcag 781 atattgattg atggtcattc cagtctgctg gcgaaattgc tgacgcaaaa cgcgctcact 841 gcacgatgcc tcatcacaaa atttatccag cgcaaaggga cttttcaggc tagccgccag 901 ccgggtaatc agcttatcca gcaacgtttc gctggatgtt ggcggcaacg aatcactggt 961 gtaacgatgg cgattcagca acatcaccaa ctgcccgaac agcaactcag ccatttcgtt 1021 agcaaacggc acatgctgac tactttcatg ctcaagctga ccaataacct gccgcgcctg 1081 cgccatcccc atgctaccta agcgccagtg tggttgccct gcgctggcgt taaatcccgg 1141 aatcgccccc tgccagtcaa gattcagctt cagacgctcc gggcaataaa taatattctg 1201 caaaaccaga tcgttaacgg aagcgtagga gtgtttatca tcagcatgaa tgtaaaagag 1261 atcgccacgg gtaatgcgat aagggcgatc gttgagtaca tgcaggccat taccgcgcca 1321 gacaatcacc agctcacaaa aatcatgtgt atgttcagca aagacatctt gcggataacg 1381 gtcagccaca gcgactgcct gctggtcgct ggcaaaaaaa tcatctttga gaagttttaa 1441 ctgatgcgcc accgtggcta cctcggccag agaacgaagt tgattattcg caatatggcg 1501 tacaaatacg ttgagaagat tcgcgttatt gcagaaagcc atcccgtccc tggcgaatat 1561 cacgcggtga ccagttaaac tctcggcgaa aaagcgtcga aaagtggtta ctgtcgctga 1621 atccacagcg ataggcgatg tcagtaacgc tggcctcgct gtggcgtagc agatgtcggg 1681 ctttcatcag tcgcaggcgg ttcaggtatc gctgaggcgt cagtcccgtt tgctgcttaa 1741 gctgccgatg tagcgtacgc agtgaaagag aaaattgatc cgccacggca tcccaattca 1801 cctcatcggc aaaatggtcc tccagccagg ccagaagcaa gttgagacgt gatgcgctgt 1861 tttccaggtt ctcctgcaaa ctgcttttac gcagcaagag cagtaattgc ataaacaaga 1921 tctcgcgact ggcggtcgag ggtaaatcat tttccccttc ctgctgttcc atctgtgcaa 1981 ccagctgtcg cacctgctgc aatacgctgt ggttaacgcg ccagtgagac ggatactgcc 2041 catccagctc ttgtggcagc aactgattca gcccggcgag aaactgaaat cgatccggcg 2101 agcgatacag cacattggtc agacacagat tatcggtatg ttcatacaga tgccgatcat 2161 gatcgcgtac gaaacagacc gtgccaccgg tgatggtata gggctgccca ttaaacacat 2221 gaatacccgt gccatgttcg acaatcacaa tttcatgaaa atcatgatga tgttcaggaa 2281 aatccgcctg cgggagccgg ggttctatcg ccacggacgc gttaccagac ggaaaaaaat 2341 ccacactatg taatacggtc atactggcct cctgatgtcg tcaacacggc gaaatagtaa 2401 tcacgaggtc aggttcttac cttaaatttt cgacggaaaa ccacgtaaaa aacgtcgatt 2461 tttcaagata cagcgtgaat tttcaggaaa tgcggtgagc atcacatcac cacaattcag 2521 caaattgtga acatcatcac gttcatcttt ccctggttgc caatggccca ttttcttgtc 2581 agtaacgaga aggtcgcgaa tccaggcgct ttttagactg gtcgtatact agagattaaa 2641 gaggagaaac cctcgatgaa gaaaagaggg gcgtttttag ggctgttgtt ggtttctgcc 2701 tgcgcatcag ttttcgctgc gaataacgaa acgagcaaga gcgtcacctt cccgaagtgc 2761 gaggatctgg acgcagcggg cattgctgcc agcgtgaaac gtgactacca acagaaccgc 2821 gtggctcgtt gggcagacga ccagaagatc gtcggtcaag cggacccggt tgcctgggtt 2881 tccctgcaag atatccaggg caaagatgat aaatggagcg tcccgctgac cgttcgtggt 2941 aagagcgcgg acattcacta tcaggtgagc gtcgattgca aagccggcat ggcggagtac 3001 caacgccgcc gtaaaggcga agagctgttc actggtgtcg tccctattct ggtggaactg 3061 gatggtgatg tcaacggtca taagttttcc gtgcgtggcg agggtgaagg tgacgcaact 3121 aatggtaaac tgacgctgaa gttcatctgt actactggta aactgccggt accttggccg 3181 actctggtaa cgacgctgac ttatggtgtt cagtgctttg ctcgttatcc ggaccacatg 3241 aagcagcatg acttcttcaa gtccgccatg ccggaaggct atgtgcagga acgcacgatt 3301 tcctttaagg atgacggcac gtacaaaacg cgtgcggaag tgaaatttga aggcgatacc 3361 ctggtaaacc gcattgagct gaaaggcatt gactttaaag aagacggcaa tatcctgggc 3421 cataagctgg aatacaattt taacagccac aatgtttaca tcaccgccga taaacaaaaa 3481 aatggcatta aagcgaattt taaaattcgc cacaacgtgg aggatggcag cgtgcagctg 3541 gctgatcact accagcaaaa cactccaatc ggtgatggtc ctgttctgct gccagacaat 3601 cactatctga gcacgcaaag cgttctgtct aaagatccga acgagaaacg cgatcacatg 3661 gttctgctgg agttcgtaac cgcagcgggc atcacgcatg gtatggatga actgtacaaa 3721 tgaccaatta ttgaaggccg ctaacgcggc ctttttttgt ttctggtctc ccaaaaattt 3781 atttgcttat taatcatccg gctcgtataa tgtgtggagg gcccaagttc acttaaaaag 3841 gagaatagtt atgaaatcga acaatgcgct catcgtcatc ctcggcaccg tcaccctgga 3901 tgctgtaggc ataggcttgg ttatgccggt actgccgggc ctcttgcggg atatcgtcca 3961 ttccgacagc atcgccagtc actatggcgt gctgctagcg ctatatgcgt tgatgcaatt 4021 tctatgcgca cccgttctcg gagcactgtc cgaccgcttt ggccgccgcc cagtcctgct 4081 cgcttcgcta cttggagcca ctatcgacta cgcgatcatg gcgaccacac ccgtcctgtg 4141 gattctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg 4201 cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag 4261 cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat 4321 ctccctgcac gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg 4381 ctgcttccta atgcaggagt cgcataaggg agagcgccgt ccgatgccct tgagagcctt 4441 caacccagtc agctccttcc ggtgggcgcg gggcatgacc attgtggccg cacttatgac 4501 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg tcattttcgg 4561 cgaggaccgc tttcgctgga gcgcgacgat gagcggcctg tcgcttgcgg tattcggaat 4621 cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa 4681 gcaggccatt atcgccggca tggcggccga cgcgctgggc tacgtcttgc tggcgttcgc 4741 gacgcgaggc tggatggcct tccccattat gattcttctc gcttccggcg gcatcgggat 4801 gcccgcgttg caggccatgc tgtccaggca ggtagatgac gaccatcagg gacagcttca 4861 aggatcgctc gcggctctta ccagcctaac ttcgatcact ggaccgctga tcgtcacggc 4921 gatttatgcc gcctcggcga gcacatggaa cgggttggca tggattgtag gcgccgccct 4981 ataccttgtc tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca cctcgacctg 5041 agacaattgt ccagataaaa aaaatcctta gctttcgcta aggatgattt ctaaaatttt 5101 tctgtcatct cttcgttatt aatgtttgta attgactgaa tatcaacgct tatttaaatc 5161 agactgaaga ctttatctct ctgtcataaa actgtcatat tccttacata taactgtcac 5221 ctgtttgtcc tattttgctt ctcgtagcca acaaacaatg ctttatgaat cctcccagga 5281 gacattatga aagttatgcg taccaccgtc gcaactgttg tcgccgcgac cttatcgatg 5341 agtgctttct ctgtgtttgc agaagcaagc ctgacaggtg caggtgcaac cttccctgcg 5401 ccggtgtatg ccaaatgggc tgacacttac cagaaagaaa ccggtaataa agttaactac 5461 cagggtatcg gttcttccgg tggcgtaaaa cagattatcg ctaataccgt tgattttggt 5521 gcctctgacg cgcc //