LOCUS SEGA023 4711 bp ds-DNA linear UNA 17-JUN-2021 DEFINITION LMBP 12859. ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 4711) AUTHORS Morten Norholm Group TITLE Direct Submission JOURNAL Exported Jun 17, 2021 from SnapGene 4.2.11 COMMENT SEGA landing pad: Ptrc-low-Ubiquitin-GFP-T-tetA-S2P11-S3P22-rrnC FEATURES Location/Qualifiers source 1..4711 /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(604..1686) /codon_start=1 /gene="lacI" /product="lac repressor" /label=lacI /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" promoter complement(1687..1764) /gene="lacI" /label=lacI promoter promoter 1795..1824 /label=The trc promoter combines the -35 region from t /label=The trc promoter combines the -35 region from the /note="The trc promoter combines the -35 region from the trp promoter and the -10 region from the lacUV5 promoter" misc_feature 1795..1824 /label=trc promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" misc_feature 1795..1800 /label=-35 -35_signal 1795..1800 /gene="trp" /label=trp misc_feature 1818..1824 /label=-10 -10_signal 1818..1824 /gene="lacUV5" /label=lacUV5 misc_feature 1825..1848 /label=lacO RBS 1860..1871 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" misc_feature 1872..1886 /label=TIR CDS 1878..2177 /codon_start=1 /label=Ubiquitin /translation="MQIFVKTLTGKTITLEVEPGSAHHHHHHHHHHAGSSDTIENVKAK IQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQEKSTLELVLELQNETQSATSDA" CDS 1944..1973 /codon_start=1 /product="10xHis affinity tag" /label=10xHis /translation="HHHHHHHHHH" CDS 2187..2897 /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 2901..2949 /label=L3S3P22 Terminator (synthetic) promoter 2960..2988 /label=Promoter 3BCD2 CDS 3028..4218 /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 4199..4248 /label=HR3 terminator 4229..4269 /label=rrnC misc_feature 4270..4319 /label=homology CDS 4464..4711 /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 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc 661 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag 721 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg 781 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat 841 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg 901 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag 961 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat 1021 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt 1081 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc 1141 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg 1201 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac 1261 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat 1321 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc 1381 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat 1441 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag 1501 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc 1561 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac 1621 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg 1681 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa 1741 ggttttgcgc cattcgatgg tgtcggatcg agatcgatct cgatcccgcg aaatttgaca 1801 attaatcatc cggctcgtat aatggaattg tgagcggata acaattccta ctagagatta 1861 aagaggagaa aggggggatg caaattttcg tgaaaacact gaccggcaaa accattaccc 1921 tggaggtgga gcctggcagc gcccatcatc atcaccatca ccatcaccat catgctggta 1981 gcagcgatac gatcgaaaac gtcaaagcca aaattcagga caaagagggc attccgcctg 2041 atcaacaacg tctgatcttc gccggaaaac agctggagga tggccgtaca ctgagtgact 2101 ataacattca agagaaatcg acactggaac tggttctgga gctgcagaat gaaacccagt 2161 cagcaacttc tgacgcttct ggcggtagca aaggagaaga acttttcact ggagttgtgc 2221 caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg 2281 gtgaaggtga tgctacatac ggaaaactca cccttaaatt tatttgcact actggaaaac 2341 tacctgttcc atggccaaca cttgtcacta ctctgaccta tggcgttcaa tgcttttccc 2401 gttatccgga tcacatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg 2461 tacaggaacg cactatatct ttcaaagatg acgggaacta caagacgcgt gctgaagtca 2521 agtttgaagg tgataccctt gttaatcgta tcgagttaaa gggtattgat tttaaagaag 2581 atggaaacat tctcggacac aaactagagt acaactataa ctcacacaat gtatacatca 2641 cggcagacaa acaaaagaat ggaatcaaag ctaacttcaa aattcgccac aacattgaag 2701 atggttccgt tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg 2761 tccttttacc agacaaccat tacctgtcga cacaatctgc cctttcgaaa gatcccaacg 2821 aaaagcgtga ccacatggtc cttcttgagt ttgtaactgc tgctgggatt acacatggca 2881 tggatgagct ctacaaatga ccaattattg aaggccgcta acgcggcctt tttttgtttc 2941 tggtctccca aaaatttatt tgcttattaa tcatccggct cgtataatgt gtggagggcc 3001 caagttcact taaaaaggag aatagttatg aaatcgaaca atgcgctcat cgtcatcctc 3061 ggcaccgtca ccctggatgc tgtaggcata ggcttggtta tgccggtact gccgggcctc 3121 ttgcgggata tcgtccattc cgacagcatc gccagtcact atggcgtgct gctagcgcta 3181 tatgcgttga tgcaatttct atgcgcaccc gttctcggag cactgtccga ccgctttggc 3241 cgccgcccag tcctgctcgc ttcgctactt ggagccacta tcgactacgc gatcatggcg 3301 accacacccg tcctgtggat tctctacgcc ggacgcatcg tggccggcat caccggcgcc 3361 acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc 3421 cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg 3481 ggactgttgg gcgccatctc cctgcacgca ccattccttg cggcggcggt gctcaacggc 3541 ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga gcgccgtccg 3601 atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgaccatt 3661 gtggccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg 3721 ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgag cggcctgtcg 3781 cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc 3841 aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc gctgggctac 3901 gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat tcttctcgct 3961 tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt agatgacgac 4021 catcagggac agcttcaagg atcgctcgcg gctcttacca gcctaacttc gatcactgga 4081 ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca catggaacgg gttggcatgg 4141 attgtaggcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg tgcatggagc 4201 cgggccacct cgacctgaga caattgtcca gataaaaaaa atccttagct ttcgctaagg 4261 atgatttcta aaatttttct gtcatctctt cgttattaat gtttgtaatt gactgaatat 4321 caacgcttat ttaaatcaga ctgaagactt tatctctctg tcataaaact gtcatattcc 4381 ttacatataa ctgtcacctg tttgtcctat tttgcttctc gtagccaaca aacaatgctt 4441 tatgaatcct cccaggagac attatgaaag ttatgcgtac caccgtcgca actgttgtcg 4501 ccgcgacctt atcgatgagt gctttctctg tgtttgcaga agcaagcctg acaggtgcag 4561 gtgcaacctt ccctgcgccg gtgtatgcca aatgggctga cacttaccag aaagaaaccg 4621 gtaataaagt taactaccag ggtatcggtt cttccggtgg cgtaaaacag attatcgcta 4681 ataccgttga ttttggtgcc tctgacgcgc c //