LOCUS SEGA089 6529 bp ds-DNA linear UNA 09-SEP-2021 DEFINITION natural linear DNA. ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 6529) 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(605..1753) /codon_start=1 /gene="galK" /gene_synonym="ECK0746; galA; JW0740" /locus_tag="b0757" /product="galactokinase" /function="enzyme; Degradation of small molecules: Carbon compounds" /EC_number="2.7.1.6" /label=galK /note="galK" /db_xref="GI:16128725" /db_xref="ASAP:ABE-0002568" /db_xref="UniProtKB/Swiss-Prot:P0A6T3" /db_xref="EcoGene:EG10363" /db_xref="GeneID:945358" /protein_id="NP_415278.1" /translation="MSLKEKTQSLFANAFGYPATHTIQAPGRVNLIGEHTDYNDGFVLP CAIDYQTVISCAPRDDRKVRVMAADYENQLDEFSLDAPIVAHENYQWANYVRGVVKHLQ LRNNSFGGVDMVISGNVPQGAGLSSSASLEVAVGTVLQQLYHLPLDGAQIALNGQEAEN QFVGCNCGIMDQLISALGKKDHALLIDCRSLGTKAVSMPKGVAVVIINSNFKRTLVGSE YNTRREQCETGARFFQQPALRDVTIEEFNAVAHELDPIVAKRVRHILTENARTVEAASA LEQGDLKRMGELMAESHASMRDDFEITVPQIDTLVEIVKAVIGDKGGVRMTGGGFGGCI VALIPEELVPAVQQAVAEQYEAKTGIKETFYVCKPSQGAGQC" promoter complement(1783..1830) /label=EM7 promoter /note="EM7 promoter" /note="synthetic bacterial promoter" terminator 1843..1886 /label=Terminator rrnB T1 /note="Terminator rrnB T1" CDS complement(1899..2744) /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(2818..3654) /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 3800..3918 /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 3930..3956 /note="TIR" /note="high TIR" CDS 3948..3968 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /note="TEV site" /translation="MENLYFQ" misc_feature 3969..4004 /label=PROTi-Linker /note="PROTi-Linker" CDS 4005..4715 /codon_start=1 /product="green fluorescent protein" /label=GFP /note="GFP" /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGN YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF VTAAGITHGMDELYK" TATA_signal 4719..4767 /label=L3S3P22 Terminator (synthetic) /note="L3S3P22 Terminator" promoter 4778..4806 /label=Promoter 3BCD2 /note="Promoter 3BCD2" CDS 4846..6036 /codon_start=1 /gene="tet" /product="tetracycline efflux protein" /label=TcR /note="TcR" /note="confers resistance to tetracycline" /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST" terminator 6047..6087 /label=rrnC /note="rrnC" CDS 6282..6529 /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 gtccaccgta 601 ttcatcagca ctgtcctgct ccttgtgatg gtttacaaac gtaaaaagtc tctttaatac 661 ctgtttttgc ttcatattgt tcagcgacag cttgctgtac ggcaggcacc agctcttccg 721 ggatcagcgc gacgatacag ccgccaaatc cgccgccggt catgcgtacg ccacctttgt 781 cgccaatcac agctttgacg atttctacca gagtgtcaat ttgcggcacg gtgatttcga 841 aatcatcgcg catagaggca tgagactccg ccatcaactc gcccatacgt ttcaggtcgc 901 cttgctccag cgcgctggca gcttcaacgg tgcgggcgtt ttcagtcagt atatgacgca 961 cgcgttttgc cacgatcggg tccagttcat gcgcaacagc gttgaactct tcaatggtga 1021 catcacgcag ggctggctgc tggaagaaac gcgcaccggt ttcgcactgt tcacgacggg 1081 tgttgtattc gctgccaacc agggtacgtt tgaagttact gttgatgatg acgacagcca 1141 cacctttggg catggaaact gctttggtcc ccagtgagcg gcaatcgatc agcaaggcat 1201 gatctttctt gccgagcgcg gaaattagct gatccatgat cccgcagtta cagcctacaa 1261 actggttttc tgcttcctga ccgttaagcg cgatttgtgc gccgtccagc ggcagatgat 1321 aaagctgctg caatacggtt ccgaccgcga cttccagtga agcggaagaa cttaacccgg 1381 caccctgcgg cacattgccg ctgatcacca tgtccacgcc gccgaagctg ttgttacgca 1441 gttgcagatg tttcaccacg ccacgaacgt agttagccca ttgatagttt tcatgtgcga 1501 caatgggcgc atcgagggaa aactcgtcga gctgattttc ataatcggct gccatcacgc 1561 gaactttacg gtcatcgcgt ggtgcacaac tgatcacggt ttgataatca atcgcgcagg 1621 gcagaacgaa accgtcgttg tagtcggtgt gttcaccaat caaattcacg cggccaggcg 1681 cctgaatggt gtgagtggca gggtagccaa atgcgttggc aaacagagat tgtgtttttt 1741 ctttcagact catgattgcc tcctgggtta gttcctcacc ttgtcgtatt atactatgcc 1801 gatatactat gccgatgatt aattgtcaac ctttaatgaa ttataaaacg aaaggctcag 1861 tcgaaagact gggcctttcg ttttatgcgt tgcgcttaat ctttctgcga attgagatga 1921 cgccactggc tgggcgtcat cccggtttcc cgggtaaaca ccaccgaaaa atagttacta 1981 tcttcaaagc cacattcggt cgaaatatca ctgattaaca ggcggctatg ctggagaaga 2041 tattgcgcat gacacactct gacctgtcgc agatattgat tgatggtcat tccagtctgc 2101 tggcgaaatt gctgacgcaa aacgcgctca ctgcacgatg cctcatcaca aaatttatcc 2161 agcgcaaagg gacttttcag gctagccgcc agccgggtaa tcagcttatc cagcaacgtt 2221 tcgctggatg ttggcggcaa cgaatcactg gtgtaacgat ggcgattcag caacatcacc 2281 aactgcccga acagcaactc agccatttcg ttagcaaacg gcacatgctg actactttca 2341 tgctcaagct gaccaataac ctgccgcgcc tgcgccatcc ccatgctacc taagcgccag 2401 tgtggttgcc ctgcgctggc gttaaatccc ggaatcgccc cctgccagtc aagattcagc 2461 ttcagacgct ccgggcaata aataatattc tgcaaaacca gatcgttaac ggaagcgtag 2521 gagtgtttat catcagcatg aatgtaaaag agatcgccac gggtaatgcg ataagggcga 2581 tcgttgagta catgcaggcc attaccgcgc cagacaatca ccagctcaca aaaatcatgt 2641 gtatgttcag caaagacatc ttgcggataa cggtcagcca cagcgactgc ctgctggtcg 2701 ctggcaaaaa aatcatcttt gagaagtttt aactgatgcg ccaccgtggc tacctcggcc 2761 agagaacgaa gttgattatt cgcaatatgg cgtacaaata cgttgagaag attcgcgtta 2821 ttgcagaaag ccatcccgtc cctggcgaat atcacgcggt gaccagttaa actctcggcg 2881 aaaaagcgtc gaaaagtggt tactgtcgct gaatccacag cgataggcga tgtcagtaac 2941 gctggcctcg ctgtggcgta gcagatgtcg ggctttcatc agtcgcaggc ggttcaggta 3001 tcgctgaggc gtcagtcccg tttgctgctt aagctgccga tgtagcgtac gcagtgaaag 3061 agaaaattga tccgccacgg catcccaatt cacctcatcg gcaaaatggt cctccagcca 3121 ggccagaagc aagttgagac gtgatgcgct gttttccagg ttctcctgca aactgctttt 3181 acgcagcaag agcagtaatt gcataaacaa gatctcgcga ctggcggtcg agggtaaatc 3241 attttcccct tcctgctgtt ccatctgtgc aaccagctgt cgcacctgct gcaatacgct 3301 gtggttaacg cgccagtgag acggatactg cccatccagc tcttgtggca gcaactgatt 3361 cagcccggcg agaaactgaa atcgatccgg cgagcgatac agcacattgg tcagacacag 3421 attatcggta tgttcataca gatgccgatc atgatcgcgt acgaaacaga ccgtgccacc 3481 ggtgatggta tagggctgcc cattaaacac atgaataccc gtgccatgtt cgacaatcac 3541 aatttcatga aaatcatgat gatgttcagg aaaatccgcc tgcgggagcc ggggttctat 3601 cgccacggac gcgttaccag acggaaaaaa atccacacta tgtaatacgg tcatactggc 3661 ctcctgatgt cgtcaacacg gcgaaatagt aatcacgagg tcaggttctt accttaaatt 3721 ttcgacggaa aaccacgtaa aaaacgtcga tttttcaaga tacagcgtga attttcagga 3781 aatgcggtga gcatcacatc accacaattc agcaaattgt gaacatcatc acgttcatct 3841 ttccctggtt gccaatggcc cattttcttg tcagtaacga gaaggtcgcg aatccaggcg 3901 ctttttagac tggtcgtata ctagagatta aagaggagaa aatatcgatg gagaacctgt 3961 acttccagtt tcgtagcaaa ggagaagaac ttgtcactgg agttagcaaa ggagaagaac 4021 ttttcactgg agttgtgcca attcttgttg aattagatgg tgatgttaat gggcacaaat 4081 tttctgtcag tggagagggt gaaggtgatg ctacatacgg aaaactcacc cttaaattta 4141 tttgcactac tggaaaacta cctgttccat ggccaacact tgtcactact ctgacctatg 4201 gcgttcaatg cttttcccgt tatccggatc acatgaaacg gcatgacttt ttcaagagtg 4261 ccatgcccga aggttatgta caggaacgca ctatatcttt caaagatgac gggaactaca 4321 agacgcgtgc tgaagtcaag tttgaaggtg atacccttgt taatcgtatc gagttaaagg 4381 gtattgattt taaagaagat ggaaacattc tcggacacaa actagagtac aactataact 4441 cacacaatgt atacatcacg gcagacaaac aaaagaatgg aatcaaagct aacttcaaaa 4501 ttcgccacaa cattgaagat ggttccgttc aactagcaga ccattatcaa caaaatactc 4561 caattggcga tggccctgtc cttttaccag acaaccatta cctgtcgaca caatctgccc 4621 tttcgaaaga tcccaacgaa aagcgtgacc acatggtcct tcttgagttt gtaactgctg 4681 ctgggattac acatggcatg gatgagctct acaaatgacc aattattgaa ggccgctaac 4741 gcggcctttt tttgtttctg gtctcccaaa aatttatttg cttattaatc atccggctcg 4801 tataatgtgt ggagggccca agttcactta aaaaggagaa tagttatgaa atcgaacaat 4861 gcgctcatcg tcatcctcgg caccgtcacc ctggatgctg taggcatagg cttggttatg 4921 ccggtactgc cgggcctctt gcgggatatc gtccattccg acagcatcgc cagtcactat 4981 ggcgtgctgc tagcgctata tgcgttgatg caatttctat gcgcacccgt tctcggagca 5041 ctgtccgacc gctttggccg ccgcccagtc ctgctcgctt cgctacttgg agccactatc 5101 gactacgcga tcatggcgac cacacccgtc ctgtggattc tctacgccgg acgcatcgtg 5161 gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat 5221 ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg 5281 gcaggccccg tggccggggg actgttgggc gccatctccc tgcacgcacc attccttgcg 5341 gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat 5401 aagggagagc gccgtccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg 5461 gcgcggggca tgaccattgt ggccgcactt atgactgtct tctttatcat gcaactcgta 5521 ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg 5581 acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc tcaagccttc 5641 gtcactggtc ccgccaccaa acgtttcggc gagaagcagg ccattatcgc cggcatggcg 5701 gccgacgcgc tgggctacgt cttgctggcg ttcgcgacgc gaggctggat ggccttcccc 5761 attatgattc ttctcgcttc cggcggcatc gggatgcccg cgttgcaggc catgctgtcc 5821 aggcaggtag atgacgacca tcagggacag cttcaaggat cgctcgcggc tcttaccagc 5881 ctaacttcga tcactggacc gctgatcgtc acggcgattt atgccgcctc ggcgagcaca 5941 tggaacgggt tggcatggat tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg 6001 cgtcgcggtg catggagccg ggccacctcg acctgagaca attgtccaga taaaaaaaat 6061 ccttagcttt cgctaaggat gatttctaaa atttttctgt catctcttcg ttattaatgt 6121 ttgtaattga ctgaatatca acgcttattt aaatcagact gaagacttta tctctctgtc 6181 ataaaactgt catattcctt acatataact gtcacctgtt tgtcctattt tgcttctcgt 6241 agccaacaaa caatgcttta tgaatcctcc caggagacat tatgaaagtt atgcgtacca 6301 ccgtcgcaac tgttgtcgcc gcgaccttat cgatgagtgc tttctctgtg tttgcagaag 6361 caagcctgac aggtgcaggt gcaaccttcc ctgcgccggt gtatgccaaa tgggctgaca 6421 cttaccagaa agaaaccggt aataaagtta actaccaggg tatcggttct tccggtggcg 6481 taaaacagat tatcgctaat accgttgatt ttggtgcctc tgacgcgcc //