LOCUS pTE5428_[pSB0156_1_revTU1_PT7-lacO-RBS35-TetR-T7term] 2652 bp DNA circular UNA 27-OCT-2024 DEFINITION Lvl1 MoClo vector, PT7-lacO driving tetR expression, reverse TU1, Amp/ColE1_v2-5C1SI-PT7-lacO-RBS0035-TetR-T7 terminator-3C1RSI-v2. ACCESSION urn.local.BarthelS.46q-idhjfyu VERSION urn.local.BarthelS.46q-idhjfyu KEYWORDS . SOURCE ORGANISM . FEATURES Location/Qualifiers terminator complement(14..110) /ApEinfo_fwdcolor="#FF6666" /ApEinfo_revcolor="#FF6666" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}" /label="AmpR Terminator" CDS complement(111..968) /ApEinfo_fwdcolor="#9F4240" /ApEinfo_revcolor="#9F4240" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}" /label="AmpR" primer_bind complement(116..138) /Sequence="GATAGGTGCCTCACTGATTAAGC" /Tm="58.9" /%GC="47.8" /Hairpin_Tm="34.4" /Self_Dimer_Tm="None" /created_by="BarthelS" /note="Derived using Geneious Prime 2023.1.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="oSB0022_lvl1_seq_rev" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; oSB0022_lvl1_seq_r ev" /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; oSB0022_lvl1_seq_r ev" /label="oSB0022_lvl1_seq_rev" promoter complement(969..1099) /ApEinfo_fwdcolor="#FF6666" /ApEinfo_revcolor="#FF6666" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}" /label="AmpR Promoter" rep_origin complement(1100..1863) /ApEinfo_fwdcolor="#7F7F7F" /ApEinfo_revcolor="#7F7F7F" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}" /label="ColE1" primer_bind 1383..1406 /created_by="Hoffmann-Becking" /Sequence="CCTCGCTCTGCTAATCCTGTTACC" /Tm="62.8" /%GC="54.2" /Hairpin_Tm="None" /Self_Dimer_Tm="None" /modified_by="BarthelS" /note="Derived using Geneious Prime 2024.0.7 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pTE5415 [pSB0112_0_04_phi29 DNAP wt]" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; oMHB011_qPCR_rev (pTE5415 [pSB0112_0_04_phi29 DNAP wt])" /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; oMHB011_qPCR_rev (pTE5422 [pSB0101_1_PT7-4bp-tetO_B0035-sfGFP_1xpepper_T7term])" /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; oMHB011_qPCR_rev (pTE5425 [pSB0146_0_04_phi29 DNAP (p2) [D246E catalytically inactive]])" /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; oMHB011_qPCR_rev (pTE5426 [pSB0143_PT7-tetO-B0035-phi29 DNAP-T7term])" /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; oMHB011_qPCR_rev (pTE5427 [pSB0161_1_TU2_PT7-tetO-RBS35-p2_dead-T7term])" /label="oMHB011_qPCR_rev" primer_bind complement(1522..1542) /created_by="Hoffmann-Becking" /Sequence="GGTGTAGGTCGTTCGCTCCAA" /Tm="62.4" /%GC="57.1" /Hairpin_Tm="None" /Self_Dimer_Tm="8.3" /modified_by="BarthelS" /note="Derived using Geneious Prime 2024.0.7 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pTE5415 [pSB0112_0_04_phi29 DNAP wt]" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; oMHB010_qPCR_fw (pTE5415 [pSB0112_0_04_phi29 DNAP wt])" /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; oMHB010_qPCR_fw (pTE5422 [pSB0101_1_PT7-4bp-tetO_B0035-sfGFP_1xpepper_T7term])" /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; oMHB010_qPCR_fw (pTE5425 [pSB0146_0_04_phi29 DNAP (p2) [D246E catalytically inactive]])" /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; oMHB010_qPCR_fw (pTE5426 [pSB0143_PT7-tetO-B0035-phi29 DNAP-T7term])" /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; oMHB010_qPCR_fw (pTE5427 [pSB0161_1_TU2_PT7-tetO-RBS35-p2_dead-T7term])" /label="oMHB010_qPCR_fw" primer_bind 1789..1806 /Sequence="CGGTTCCTGGCCTTTTGC" /Tm="59.3" /%GC="61.1" /Hairpin_Tm="40.2" /Self_Dimer_Tm="3.3" /created_by="BarthelS" /note="Derived using Geneious Prime 2023.1.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="oSB0021_lvl1_seq_fwd" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; oSB0021_lvl1_seq_f wd" /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; oSB0021_lvl1_seq_f wd" /label="oSB0021_lvl1_seq_fwd" promoter 1891..1908 /note="T7 RNA polymerase promoter" /regulatory_class="promoter" /note="Derived using Geneious Prime 2020.1.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="T7" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; T7" /label="T7" Operator 1914..1932 /kind="operator" /note="Lac operon operator" /note="Derived using Geneious Prime 2020.1.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="lacO" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; lacO" /modified_by="BarthelS" /label="lacO" RBS 1941..1954 /created_by="BarthelS" /note="Derived using Geneious Prime 2024.0.7 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pTE5422 [pSB0101_1_PT7-4bp-tetO_B0035-sfGFP_1xpepper_T7term]" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; B0035 (pTE5422 [pSB0101_1_PT7-4bp-tetO_B0035-sfGFP_1xpepper_T7term])" /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; B0035 (pTE5426 [pSB0143_PT7-tetO-B0035-phi29 DNAP-T7term])" /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; B0035 (pTE5427 [pSB0161_1_TU2_PT7-tetO-RBS35-p2_dead-T7term])" /label="B0035" CDS <1961..2584 /gene="tetR from transposon Tn10" /note="TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline." /product="tetracycline repressor TetR" /protein_id="
" /transl_table=1 /Original_Translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLG VEQPTLYWHVKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSH RDGAKVHLGTRPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEH QVAKEERETPTTDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS" /modified_by="sbarthel" /label="TetR" terminator 2591..2637 //note="copied from pJBL704" /created_by="sbarthel" /label="T7 terminator" ORIGIN 1 agactgcggc cgcgattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 61 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 121 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 181 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 241 ataccgcggg acccacgctc accggctcca gatttatcag caataaacca gccagccgga 301 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 361 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 421 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc 481 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc 541 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca 601 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag 661 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg 721 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa 781 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa 841 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga 901 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga 961 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg 1021 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt 1081 ccccgaaaag tgccacctgt catgaccaaa atcccttaac gtgagttttc gttccactga 1141 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta 1201 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa 1261 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact 1321 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca 1381 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt 1441 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg 1501 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag 1561 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta 1621 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat 1681 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg 1741 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc 1801 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac 1861 cgtgcggccg caacacgtct cgagtaggag taatacgact cactataggg gaattgtgag 1921 cggataacaa tttactagag attaaagagg agaataatca atgtctagat tagataaaag 1981 taaagtgatt aacagcgcat tagagctgct taatgaggtc ggaatcgaag gtttaacaac 2041 ccgtaaactc gcccagaagc taggtgtaga gcagcctaca ttgtattggc atgtaaaaaa 2101 taagcgggct ttgctcgacg ccttagccat tgagatgtta gataggcacc atactcactt 2161 ttgcccttta gaaggggaaa gctggcaaga ttttttacgt aataacgcta aaagttttag 2221 atgtgcttta ctaagtcatc gcgatggagc aaaagtacat ttaggtacac ggcctacaga 2281 aaaacagtat gaaactctcg aaaatcaatt agccttttta tgccaacaag gtttttcact 2341 agagaatgca ttatatgcac tcagcgctgt ggggcatttt actttaggtt gcgtattgga 2401 agatcaagag catcaagtcg ctaaagaaga aagggaaaca cctactactg atagtatgcc 2461 gccattatta cgacaagcta tcgaattatt tgatcaccaa ggtgcagagc cagccttctt 2521 attcggcctt gaattgatca tatgcggatt agagaagcaa cttaagtgtg aaagtgggtc 2581 ttaagcttaa tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgcgc 2641 ttgttggaga cg //