LOCUS pTE5427_[pSB0161_1_TU2_PT7-tetO-RBS35-p2_dead-T7term] 3745 bp DNA circular UNA 27-OCT-2024 DEFINITION Lvl1 MoClo vector, PT7-tetO-driving expression of p2_dead, RBS: B0035, TU2-N, (Amp/ColE1_v2-5C2SN-T7-tetO-RBS0035-phi29 DNAP (p2) 'dead'-T7 terminator-3C5OSN_v2). ACCESSION urn.local.BarthelS.46p-idhjfyu VERSION urn.local.BarthelS.46p-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])" /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])" /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..1907 /created_by="sbarthel" /X-Transferred_From="T7-lacO" /X-Transferred_Similarity="100.00%" /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0; T7 phage promoter (T7-lacO)" /modified_by="BarthelS" /note="Derived using Geneious Prime 2020.2.4 'Annotate from Database' based on nucleotide similarity" /Transferred_From="T7-tetO_2bp spacer" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; T7 phage promoter (T7-tetO_2bp spacer)" /label="T7 phage promoter" Operator 1912..1930 /created_by="BarthelS" /modified_by="BarthelS" /note="Derived using Geneious Prime 2020.2.4 'Annotate from Database' based on nucleotide similarity" /Transferred_From="T7-tetO_2bp spacer" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; tetO (T7-tetO_2bp spacer)" /label="tetO" RBS 1939..1952 /created_by="BarthelS" /label="B0035" CDS <1959..3677 /modified_by="BarthelS" /label="phi29 DNAP D249E (inactive)" Mutation 2694..2696 /created_by="BarthelS" /label="D249E --> inactive" terminator 3684..3730 //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 cgtactggag taatacgact cactataggg atccctatca 1921 gtgatagaga tactagagat taaagaggag aataatcaat gccgcgtaaa atgtacagct 1981 gcgattttga aacgacgacg aaagttgaag attgccgtgt ctgggcctat ggttatatga 2041 acatcgaaga ccattcagaa tataaaattg gcaactcgct ggatgaattt atggcgtggg 2101 tgctgaaagt tcaggccgac ctgtacttcc acaatctgaa atttgatggt gcgttcatta 2161 tcaactggct ggaacgtaat ggctttaaat ggagcgccga tggtctgccg aacacctata 2221 atacgattat ctctcgtatg ggccaatggt atatgattga tatctgcctg ggctacaaag 2281 gtaaacgcaa aattcatacc gtgatctatg acagcctgaa aaaactgccg tttccggtga 2341 agaaaattgc gaaagatttc aaactgaccg tcctgaaagg cgatattgac tatcacaaag 2401 aacgtccggt tggttacaaa atcacgccgg aagaatatgc gtacattaaa aacgatatcc 2461 agattatcgc agaagctctg ctgattcagt ttaaacaagg cctggatcgc atgaccgccg 2521 gcagtgactc cctgaaaggt ttcaaagata tcatcaccac gaaaaaattt aagaaagtgt 2581 tcccgaccct gagcctgggt ctggataaag aagttcgtta tgcataccgc ggcggtttta 2641 cgtggctgaa cgaccgtttc aaagaaaaag aaattggcga gggtatggtc tttgaagtga 2701 atagtctgta tccggctcag atgtactccc gcctgctgcc gtatggcgaa ccgatcgttt 2761 tcgagggtaa atatgtctgg gatgaagact acccgctgca tattcagcac atccgttgtg 2821 aatttgaact gaaagaaggc tatattccga ccattcaaat caaacgtagc cgcttctata 2881 agggtaacga atacctgaaa agctctggcg gtgaaatcgc agacctgtgg ctgagtaacg 2941 tcgatctgga actgatgaaa gaacattacg atctgtacaa cgttgaatac atctccggcc 3001 tgaaatttaa agccaccacg ggtctgttta aagatttcat tgacaaatgg acctacatca 3061 aaaccacgtc tgaaggtgca atcaaacagc tggctaaact gatgctgaac agcctgtatg 3121 gcaaatttgc atctaatccg gatgttaccg gtaaagtccc gtacctgaaa gaaaatggcg 3181 ctctgggttt tcgcctgggc gaagaagaaa ccaaagatcc ggtgtatacg ccgatgggtg 3241 ttttcattac cgcgtgggca cgttacacca ccatcaccgc cgcacaagcg tgctatgacc 3301 gcattatcta ctgtgatacc gactcaattc atctgaccgg cacggaaatc ccggatgtga 3361 ttaaagatat cgttgacccg aaaaaactgg gttattgggc acacgaatcg acctttaaac 3421 gtgctaaata cctgcgccag aaaacgtaca tccaagacat ctacatgaaa gaagtcgatg 3481 gcaaactggt ggaaggttca ccggatgact ataccgacat taaattttcg gtgaaatgcg 3541 ccggcatgac cgataaaatt aagaaagaag tgacgttcga aaatttcaaa gtgggtttca 3601 gtcgcaaaat gaaaccgaaa ccggtccaag ttccgggcgg cgttgtgctg gtcgatgaca 3661 ccttcacgat caaataagct taatagcata accccttggg gcctctaaac gggtcttgag 3721 gggttttttg cgctagacgg agacg //