LOCUS SEGA061 4701 bp ds-DNA linear UNA 17-JUN-2021 DEFINITION LMBP 12897. ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 4701) AUTHORS Morten Norholm Group TITLE Direct Submission JOURNAL Exported Jun 17, 2021 from SnapGene 4.2.11 COMMENT SEGA landing pad: PT7-high-Ubiquitin-GFP-T-tetA-S2P11-S3P22-rrnC FEATURES Location/Qualifiers source 1..4701 /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..1813 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 1814..1838 /label=lac operator /bound_moiety="lac repressor encoded by 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)." RBS 1850..1861 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" misc_feature 1862..1876 /label=TIR CDS 1868..2167 /codon_start=1 /label=Ubiquitin /translation="MQIFVKTLTGKTITLEVEPGSAHHHHHHHHHHAGSSDTIENVKAK IQDKEGIPPDQQRLIFAGKQLEDGRTLSDYNIQEKSTLELVLELQNETQSATSDA" CDS 1934..1963 /codon_start=1 /product="10xHis affinity tag" /label=10xHis /translation="HHHHHHHHHH" CDS 2177..2887 /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 2891..2939 /label=L3S3P22 Terminator (synthetic) promoter 2950..2978 /label=Promoter 3BCD2 CDS 3018..4208 /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 4189..4238 /label=HR3 terminator 4219..4259 /label=rrnC misc_feature 4260..4309 /label=homology CDS 4454..4701 /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 aaattaatac 1801 gactcactat aggggaattg tgagcggata acaattccta ctagagatta aagaggagaa 1861 acattgtatg cagatattcg tgaaaacact gaccggcaaa accattaccc tggaggtgga 1921 gcctggcagc gcccatcatc atcaccatca ccatcaccat catgctggta gcagcgatac 1981 gatcgaaaac gtcaaagcca aaattcagga caaagagggc attccgcctg atcaacaacg 2041 tctgatcttc gccggaaaac agctggagga tggccgtaca ctgagtgact ataacattca 2101 agagaaatcg acactggaac tggttctgga gctgcagaat gaaacccagt cagcaacttc 2161 tgacgcttct ggcggtagca aaggagaaga acttttcact ggagttgtgc caattcttgt 2221 tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga 2281 tgctacatac ggaaaactca cccttaaatt tatttgcact actggaaaac tacctgttcc 2341 atggccaaca cttgtcacta ctctgaccta tggcgttcaa tgcttttccc gttatccgga 2401 tcacatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaacg 2461 cactatatct ttcaaagatg acgggaacta caagacgcgt gctgaagtca agtttgaagg 2521 tgataccctt gttaatcgta tcgagttaaa gggtattgat tttaaagaag atggaaacat 2581 tctcggacac aaactagagt acaactataa ctcacacaat gtatacatca cggcagacaa 2641 acaaaagaat ggaatcaaag ctaacttcaa aattcgccac aacattgaag atggttccgt 2701 tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc 2761 agacaaccat tacctgtcga cacaatctgc cctttcgaaa gatcccaacg aaaagcgtga 2821 ccacatggtc cttcttgagt ttgtaactgc tgctgggatt acacatggca tggatgagct 2881 ctacaaatga ccaattattg aaggccgcta acgcggcctt tttttgtttc tggtctccca 2941 aaaatttatt tgcttattaa tcatccggct cgtataatgt gtggagggcc caagttcact 3001 taaaaaggag aatagttatg aaatcgaaca atgcgctcat cgtcatcctc ggcaccgtca 3061 ccctggatgc tgtaggcata ggcttggtta tgccggtact gccgggcctc ttgcgggata 3121 tcgtccattc cgacagcatc gccagtcact atggcgtgct gctagcgcta tatgcgttga 3181 tgcaatttct atgcgcaccc gttctcggag cactgtccga ccgctttggc cgccgcccag 3241 tcctgctcgc ttcgctactt ggagccacta tcgactacgc gatcatggcg accacacccg 3301 tcctgtggat tctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg 3361 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc 3421 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg 3481 gcgccatctc cctgcacgca ccattccttg cggcggcggt gctcaacggc ctcaacctac 3541 tactgggctg cttcctaatg caggagtcgc ataagggaga gcgccgtccg atgcccttga 3601 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgaccatt gtggccgcac 3661 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca 3721 ttttcggcga ggaccgcttt cgctggagcg cgacgatgag cggcctgtcg cttgcggtat 3781 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg 3841 gcgagaagca ggccattatc gccggcatgg cggccgacgc gctgggctac gtcttgctgg 3901 cgttcgcgac gcgaggctgg atggccttcc ccattatgat tcttctcgct tccggcggca 3961 tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt agatgacgac catcagggac 4021 agcttcaagg atcgctcgcg gctcttacca gcctaacttc gatcactgga ccgctgatcg 4081 tcacggcgat ttatgccgcc tcggcgagca catggaacgg gttggcatgg attgtaggcg 4141 ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg tgcatggagc cgggccacct 4201 cgacctgaga caattgtcca gataaaaaaa atccttagct ttcgctaagg atgatttcta 4261 aaatttttct gtcatctctt cgttattaat gtttgtaatt gactgaatat caacgcttat 4321 ttaaatcaga ctgaagactt tatctctctg tcataaaact gtcatattcc ttacatataa 4381 ctgtcacctg tttgtcctat tttgcttctc gtagccaaca aacaatgctt tatgaatcct 4441 cccaggagac attatgaaag ttatgcgtac caccgtcgca actgttgtcg ccgcgacctt 4501 atcgatgagt gctttctctg tgtttgcaga agcaagcctg acaggtgcag gtgcaacctt 4561 ccctgcgccg gtgtatgcca aatgggctga cacttaccag aaagaaaccg gtaataaagt 4621 taactaccag ggtatcggtt cttccggtgg cgtaaaacag attatcgcta ataccgttga 4681 ttttggtgcc tctgacgcgc c //