LOCUS SEGA045 5201 bp ds-DNA linear UNA 08-SEP-2021 DEFINITION natural linear DNA. ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 5201) AUTHORS Morten Norholm Group TITLE Direct Submission JOURNAL Exported Sep 9, 2021 from SnapGene 4.2.11 http://www.snapgene.com FEATURES Location/Qualifiers CDS 1..528 /codon_start=1 /label=glmS /note="glmS" /translation="VHGLQALPSRIEQMLSQDKRIEALAEDFSDKHHALFLGRGDQYPI ALEGALKLKEISYIHAEAYAAGELKHGPLALIDADMPVIVVAPNNELLEKLKSNIEEVR ARGGQLYVFADQDAGFVSSDNMHIIEMPHVEEVIAPIFYTVPLQLLAYHVALIKGTDVD QPRNLAKSVTVE" terminator 538..594 /label=L3S2P11 Terminator /note="L3S2P11 Terminator" CDS complement(607..1452) /codon_start=1 /gene="E. coli rhaR" /product="transcriptional activator of rhaR and rhaS" /label=rhaR /note="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="rhaS" /note="activated by L-rhamnose" /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ" promoter 2508..2626 /label=rhaB promoter /note="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..2664 /note="TIR" /note="medium TIR" CDS 2656..2676 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /note="TEV site" /translation="MENLYFQ" CDS 2677..3387 /codon_start=1 /product="greeen fluorescent protein" /label=GFP /note="GFP" /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGN YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF VTAAGITHGMDELYK" terminator 3391..3439 /label=L3S3P22 Terminator (synthetic) /note="L3S3P22 Terminator" promoter 3450..3478 /label=Promoter 3BCD2 /note="Promoter 3BCD2" CDS 3518..4708 /codon_start=1 /gene="tet" /product="tetracycline efflux protein" /label=TcR /note="TcR" /note="confers resistance to tetracycline" /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM QLVGQVPAALWVIFGEDRFRWSATMSGLSLAVFGILHALAQAFVTGPATKRFGEKQAII AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST" terminator 4719..4759 /label=rrnC /note="rrnC" CDS 4954..5201 /codon_start=1 /label=pstS /note="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 gaggagaaaa actacatgga aaacctgtac ttccagagca aaggagaaga acttttcact 2701 ggagttgtgc caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc 2761 agtggagagg gtgaaggtga tgctacatac ggaaaactca cccttaaatt tatttgcact 2821 actggaaaac tacctgttcc atggccaaca cttgtcacta ctctgaccta tggcgttcaa 2881 tgcttttccc gttatccgga tcacatgaaa cggcatgact ttttcaagag tgccatgccc 2941 gaaggttatg tacaggaacg cactatatct ttcaaagatg acgggaacta caagacgcgt 3001 gctgaagtca agtttgaagg tgataccctt gttaatcgta tcgagttaaa gggtattgat 3061 tttaaagaag atggaaacat tctcggacac aaactagagt acaactataa ctcacacaat 3121 gtatacatca cggcagacaa acaaaagaat ggaatcaaag ctaacttcaa aattcgccac 3181 aacattgaag atggttccgt tcaactagca gaccattatc aacaaaatac tccaattggc 3241 gatggccctg tccttttacc agacaaccat tacctgtcga cacaatctgc cctttcgaaa 3301 gatcccaacg aaaagcgtga ccacatggtc cttcttgagt ttgtaactgc tgctgggatt 3361 acacatggca tggatgagct ctacaaatga ccaattattg aaggccgcta acgcggcctt 3421 tttttgtttc tggtctccca aaaatttatt tgcttattaa tcatccggct cgtataatgt 3481 gtggagggcc caagttcact taaaaaggag aatagttatg aaatcgaaca atgcgctcat 3541 cgtcatcctc ggcaccgtca ccctggatgc tgtaggcata ggcttggtta tgccggtact 3601 gccgggcctc ttgcgggata tcgtccattc cgacagcatc gccagtcact atggcgtgct 3661 gctagcgcta tatgcgttga tgcaatttct atgcgcaccc gttctcggag cactgtccga 3721 ccgctttggc cgccgcccag tcctgctcgc ttcgctactt ggagccacta tcgactacgc 3781 gatcatggcg accacacccg tcctgtggat tctctacgcc ggacgcatcg tggccggcat 3841 caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga 3901 tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc 3961 cgtggccggg ggactgttgg gcgccatctc cctgcacgca ccattccttg cggcggcggt 4021 gctcaacggc ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga 4081 gcgccgtccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg 4141 catgaccatt gtggccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt 4201 gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgag 4261 cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg 4321 tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc 4381 gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat 4441 tcttctcgct tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt 4501 agatgacgac catcagggac agcttcaagg atcgctcgcg gctcttacca gcctaacttc 4561 gatcactgga ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca catggaacgg 4621 gttggcatgg attgtaggcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg 4681 tgcatggagc cgggccacct cgacctgaga caattgtcca gataaaaaaa atccttagct 4741 ttcgctaagg atgatttcta aaatttttct gtcatctctt cgttattaat gtttgtaatt 4801 gactgaatat caacgcttat ttaaatcaga ctgaagactt tatctctctg tcataaaact 4861 gtcatattcc ttacatataa ctgtcacctg tttgtcctat tttgcttctc gtagccaaca 4921 aacaatgctt tatgaatcct cccaggagac attatgaaag ttatgcgtac caccgtcgca 4981 actgttgtcg ccgcgacctt atcgatgagt gctttctctg tgtttgcaga agcaagcctg 5041 acaggtgcag gtgcaacctt ccctgcgccg gtgtatgcca aatgggctga cacttaccag 5101 aaagaaaccg gtaataaagt taactaccag ggtatcggtt cttccggtgg cgtaaaacag 5161 attatcgcta ataccgttga ttttggtgcc tctgacgcgc c //