LOCUS SEGA029 4459 bp ds-DNA linear UNA 17-JUN-2021 DEFINITION LMBP 12865. ACCESSION . VERSION . KEYWORDS . SOURCE natural DNA sequence ORGANISM unspecified REFERENCE 1 (bases 1 to 4459) AUTHORS Morten Norholm Group TITLE Direct Submission JOURNAL Exported Jun 17, 2021 from SnapGene 4.2.11 COMMENT SEGA landing pad: Ptrc-low-PROTi-GFP-T-tetA-S2P11-S3P22-rrnC FEATURES Location/Qualifiers source 1..4459 /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..1898 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="MENLYFQ" misc_feature 1899..1934 /label=PROTi-Linker CDS 1935..2645 /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 2649..2697 /label=L3S3P22 Terminator (synthetic) promoter 2708..2736 /label=Promoter 3BCD2 CDS 2776..3966 /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 3947..3996 /label=HR3 terminator 3977..4017 /label=rrnC misc_feature 4018..4067 /label=homology CDS 4212..4459 /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 acgggggatg gagaacctgt acttccagtt tcgtagcaaa ggagaagaac 1921 ttgtcactgg agttagcaaa ggagaagaac ttttcactgg agttgtgcca attcttgttg 1981 aattagatgg tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg 2041 ctacatacgg aaaactcacc cttaaattta tttgcactac tggaaaacta cctgttccat 2101 ggccaacact tgtcactact ctgacctatg gcgttcaatg cttttcccgt tatccggatc 2161 acatgaaacg gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaacgca 2221 ctatatcttt caaagatgac gggaactaca agacgcgtgc tgaagtcaag tttgaaggtg 2281 atacccttgt taatcgtatc gagttaaagg gtattgattt taaagaagat ggaaacattc 2341 tcggacacaa actagagtac aactataact cacacaatgt atacatcacg gcagacaaac 2401 aaaagaatgg aatcaaagct aacttcaaaa ttcgccacaa cattgaagat ggttccgttc 2461 aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag 2521 acaaccatta cctgtcgaca caatctgccc tttcgaaaga tcccaacgaa aagcgtgacc 2581 acatggtcct tcttgagttt gtaactgctg ctgggattac acatggcatg gatgagctct 2641 acaaatgacc aattattgaa ggccgctaac gcggcctttt tttgtttctg gtctcccaaa 2701 aatttatttg cttattaatc atccggctcg tataatgtgt ggagggccca agttcactta 2761 aaaaggagaa tagttatgaa atcgaacaat gcgctcatcg tcatcctcgg caccgtcacc 2821 ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt gcgggatatc 2881 gtccattccg acagcatcgc cagtcactat ggcgtgctgc tagcgctata tgcgttgatg 2941 caatttctat gcgcacccgt tctcggagca ctgtccgacc gctttggccg ccgcccagtc 3001 ctgctcgctt cgctacttgg agccactatc gactacgcga tcatggcgac cacacccgtc 3061 ctgtggattc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt 3121 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc 3181 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc 3241 gccatctccc tgcacgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta 3301 ctgggctgct tcctaatgca ggagtcgcat aagggagagc gccgtccgat gcccttgaga 3361 gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgaccattgt ggccgcactt 3421 atgactgtct tctttatcat gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt 3481 ttcggcgagg accgctttcg ctggagcgcg acgatgagcg gcctgtcgct tgcggtattc 3541 ggaatcttgc acgccctcgc tcaagccttc gtcactggtc ccgccaccaa acgtttcggc 3601 gagaagcagg ccattatcgc cggcatggcg gccgacgcgc tgggctacgt cttgctggcg 3661 ttcgcgacgc gaggctggat ggccttcccc attatgattc ttctcgcttc cggcggcatc 3721 gggatgcccg cgttgcaggc catgctgtcc aggcaggtag atgacgacca tcagggacag 3781 cttcaaggat cgctcgcggc tcttaccagc ctaacttcga tcactggacc gctgatcgtc 3841 acggcgattt atgccgcctc ggcgagcaca tggaacgggt tggcatggat tgtaggcgcc 3901 gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg ggccacctcg 3961 acctgagaca attgtccaga taaaaaaaat ccttagcttt cgctaaggat gatttctaaa 4021 atttttctgt catctcttcg ttattaatgt ttgtaattga ctgaatatca acgcttattt 4081 aaatcagact gaagacttta tctctctgtc ataaaactgt catattcctt acatataact 4141 gtcacctgtt tgtcctattt tgcttctcgt agccaacaaa caatgcttta tgaatcctcc 4201 caggagacat tatgaaagtt atgcgtacca ccgtcgcaac tgttgtcgcc gcgaccttat 4261 cgatgagtgc tttctctgtg tttgcagaag caagcctgac aggtgcaggt gcaaccttcc 4321 ctgcgccggt gtatgccaaa tgggctgaca cttaccagaa agaaaccggt aataaagtta 4381 actaccaggg tatcggttct tccggtggcg taaaacagat tatcgctaat accgttgatt 4441 ttggtgcctc tgacgcgcc //