LOCUS pTE5452_[PT7-tetO-B0035-10xHis-phi29_DNAP_dead-T7_term] 3817 bp DNA circular UNA 22-NOV-2024 DEFINITION Lvl1 MoClo vector, PT7-tetO driving production of 10xHis-phi29 DNAP dead, RBS: B0035, TU2-N (5C2SN-PT7-tetO-RBS0035-10xHis-phi29 DNAP PovSak-T7term-3C2SN-AmpR-ColE1). ACCESSION urn.local.barthels.52-ihb2ndz VERSION urn.local.barthels.52-ihb2ndz 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 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 /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" unsure 1908..1911 /created_by="BarthelS" /modified_by="BarthelS" /label="4 bp spacer" 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..3749 /created_by="BarthelS" /modified_by="BarthelS" /label="10xphi29 DNAP 'dead'" misc_feature 1965..1994 /kind="tag" /note="6X polyhistidine tag used for protein purification" /Color="#FFFFB9" /note="Derived using Geneious Prime 2020.1.2 'Annotate from Database' based on protein similarity" /Transferred_From="polyhistidine" /Transferred_Translation_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; polyhistidine" /Alternative_Match="100.00%; Adjusted-Interval-0:3/3; polyhistidine" /label="polyhistidine" misc_feature 2007..2018 /kind="recognition site" /note="factor Xa recognition and cleavage site" /note="Derived using Geneious Prime 2020.1.2 'Annotate from Database' based on protein similarity" /Transferred_From="factor XA site" /Transferred_Translation_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; factor XA site" /label="factor XA site" unsure 2766..2768 /created_by="BarthelS" /modified_by="BarthelS" /label="D249E" terminator 3756..3802 //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 gggccatcat catcatcatc 1981 atcatcatca tcacagcagc ggccatatcg aaggtcgtca tggctctcga atgccgcgta 2041 aaatgtacag ctgcgatttt gaaacgacga cgaaagttga agattgccgt gtctgggcct 2101 atggttatat gaacatcgaa gaccattcag aatataaaat tggcaactcg ctggatgaat 2161 ttatggcgtg ggtgctgaaa gttcaggccg acctgtactt ccacaatctg aaatttgatg 2221 gtgcgttcat tatcaactgg ctggaacgta atggctttaa atggagcgcc gatggtctgc 2281 cgaacaccta taatacgatt atctctcgta tgggccaatg gtatatgatt gatatctgcc 2341 tgggctacaa aggtaaacgc aaaattcata ccgtgatcta tgacagcctg aaaaaactgc 2401 cgtttccggt gaagaaaatt gcgaaagatt tcaaactgac cgtcctgaaa ggcgatattg 2461 actatcacaa agaacgtccg gttggttaca aaatcacgcc ggaagaatat gcgtacatta 2521 aaaacgatat ccagattatc gcagaagctc tgctgattca gtttaaacaa ggcctggatc 2581 gcatgaccgc cggcagtgac tccctgaaag gtttcaaaga tatcatcacc acgaaaaaat 2641 ttaagaaagt gttcccgacc ctgagcctgg gtctggataa agaagttcgt tatgcatacc 2701 gcggcggttt tacgtggctg aacgaccgtt tcaaagaaaa agaaattggc gagggtatgg 2761 tctttgaagt gaatagtctg tatccggctc agatgtactc ccgcctgctg ccgtatggcg 2821 aaccgatcgt tttcgagggt aaatatgtct gggatgaaga ctacccgctg catattcagc 2881 acatccgttg tgaatttgaa ctgaaagaag gctatattcc gaccattcaa atcaaacgta 2941 gccgcttcta taagggtaac gaatacctga aaagctctgg cggtgaaatc gcagacctgt 3001 ggctgagtaa cgtcgatctg gaactgatga aagaacatta cgatctgtac aacgttgaat 3061 acatctccgg cctgaaattt aaagccacca cgggtctgtt taaagatttc attgacaaat 3121 ggacctacat caaaaccacg tctgaaggtg caatcaaaca gctggctaaa ctgatgctga 3181 acagcctgta tggcaaattt gcatctaatc cggatgttac cggtaaagtc ccgtacctga 3241 aagaaaatgg cgctctgggt tttcgcctgg gcgaagaaga aaccaaagat ccggtgtata 3301 cgccgatggg tgttttcatt accgcgtggg cacgttacac caccatcacc gccgcacaag 3361 cgtgctatga ccgcattatc tactgtgata ccgactcaat tcatctgacc ggcacggaaa 3421 tcccggatgt gattaaagat atcgttgacc cgaaaaaact gggttattgg gcacacgaat 3481 cgacctttaa acgtgctaaa tacctgcgcc agaaaacgta catccaagac atctacatga 3541 aagaagtcga tggcaaactg gtggaaggtt caccggatga ctataccgac attaaatttt 3601 cggtgaaatg cgccggcatg accgataaaa ttaagaaaga agtgacgttc gaaaatttca 3661 aagtgggttt cagtcgcaaa atgaaaccga aaccggtcca agttccgggc ggcgttgtgc 3721 tggtcgatga caccttcacg atcaaataag cttaatagca taaccccttg gggcctctaa 3781 acgggtcttg aggggttttt tgcgctagac ggagacg //