LOCUS SEGA038 5236 bp ds-DNA linear UNA 17-JUN-2021 DEFINITION LMBP 12874. ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 5236) AUTHORS Morten Norholm Group TITLE Direct Submission JOURNAL Exported Jun 17, 2021 from SnapGene 4.2.11 COMMENT SEGA landing pad: PRha-low-BCD-GFP-T-tetA-S2P11-S3P22-rrnC FEATURES Location/Qualifiers source 1..5236 /organism="unspecified" /mol_type="genomic DNA" 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 CDS 2656..2711 /codon_start=1 /label=BCD /translation="MKAIFVLKHLNHAKEVF" misc_feature 2694..2702 /label=SD2 misc_feature 2707..2709 /label=STOP BCD misc_feature 2709..2711 /label=start GOI CDS 2712..3422 /codon_start=1 /product="GFP variant that folds robustly even when fused to poorly folded proteins (Pedelacq et al., 2006) ?" /label=GFP /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGN YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF VTAAGITHGMDELYK" misc_feature 3426..3474 /label=L3S3P22 Terminator (synthetic) promoter 3485..3513 /label=Promoter 3BCD2 CDS 3553..4743 /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 4724..4773 /label=HR3 terminator 4754..4794 /label=rrnC misc_feature 4795..4844 /label=homology CDS 4989..5236 /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 gaggagaaag gggggatgaa ggctattttc gtactgaaac atcttaatca tgctaaggag 2701 gttttctaat gagcaaagga gaagaacttt tcactggagt tgtgccaatt cttgttgaat 2761 tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcta 2821 catacggaaa actcaccctt aaatttattt gcactactgg aaaactacct gttccatggc 2881 caacacttgt cactactctg acctatggcg ttcaatgctt ttcccgttat ccggatcaca 2941 tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaacgcacta 3001 tatctttcaa agatgacggg aactacaaga cgcgtgctga agtcaagttt gaaggtgata 3061 cccttgttaa tcgtatcgag ttaaagggta ttgattttaa agaagatgga aacattctcg 3121 gacacaaact agagtacaac tataactcac acaatgtata catcacggca gacaaacaaa 3181 agaatggaat caaagctaac ttcaaaattc gccacaacat tgaagatggt tccgttcaac 3241 tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca 3301 accattacct gtcgacacaa tctgcccttt cgaaagatcc caacgaaaag cgtgaccaca 3361 tggtccttct tgagtttgta actgctgctg ggattacaca tggcatggat gagctctaca 3421 aatgaccaat tattgaaggc cgctaacgcg gccttttttt gtttctggtc tcccaaaaat 3481 ttatttgctt attaatcatc cggctcgtat aatgtgtgga gggcccaagt tcacttaaaa 3541 aggagaatag ttatgaaatc gaacaatgcg ctcatcgtca tcctcggcac cgtcaccctg 3601 gatgctgtag gcataggctt ggttatgccg gtactgccgg gcctcttgcg ggatatcgtc 3661 cattccgaca gcatcgccag tcactatggc gtgctgctag cgctatatgc gttgatgcaa 3721 tttctatgcg cacccgttct cggagcactg tccgaccgct ttggccgccg cccagtcctg 3781 ctcgcttcgc tacttggagc cactatcgac tacgcgatca tggcgaccac acccgtcctg 3841 tggattctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct 3901 ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg 3961 agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact gttgggcgcc 4021 atctccctgc acgcaccatt ccttgcggcg gcggtgctca acggcctcaa cctactactg 4081 ggctgcttcc taatgcagga gtcgcataag ggagagcgcc gtccgatgcc cttgagagcc 4141 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ccattgtggc cgcacttatg 4201 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc 4261 ggcgaggacc gctttcgctg gagcgcgacg atgagcggcc tgtcgcttgc ggtattcgga 4321 atcttgcacg ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag 4381 aagcaggcca ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc 4441 gcgacgcgag gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg 4501 atgcccgcgt tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt 4561 caaggatcgc tcgcggctct taccagccta acttcgatca ctggaccgct gatcgtcacg 4621 gcgatttatg ccgcctcggc gagcacatgg aacgggttgg catggattgt aggcgccgcc 4681 ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat ggagccgggc cacctcgacc 4741 tgagacaatt gtccagataa aaaaaatcct tagctttcgc taaggatgat ttctaaaatt 4801 tttctgtcat ctcttcgtta ttaatgtttg taattgactg aatatcaacg cttatttaaa 4861 tcagactgaa gactttatct ctctgtcata aaactgtcat attccttaca tataactgtc 4921 acctgtttgt cctattttgc ttctcgtagc caacaaacaa tgctttatga atcctcccag 4981 gagacattat gaaagttatg cgtaccaccg tcgcaactgt tgtcgccgcg accttatcga 5041 tgagtgcttt ctctgtgttt gcagaagcaa gcctgacagg tgcaggtgca accttccctg 5101 cgccggtgta tgccaaatgg gctgacactt accagaaaga aaccggtaat aaagttaact 5161 accagggtat cggttcttcc ggtggcgtaa aacagattat cgctaatacc gttgattttg 5221 gtgcctctga cgcgcc //