LOCUS pTE5444_[PT7-lacO-g19lead-10xHisTag-phi29_DNAP_PovSak] 5524 bp DNA circular UNA 22-NOV-2024 DEFINITION Expression vector for N-terminal His-tagged phi29 DNA polymerase PovSak. ACCESSION urn.local.barthels.z-ihb0yg2 VERSION urn.local.barthels.z-ihb0yg2 KEYWORDS . SOURCE ORGANISM . FEATURES Location/Qualifiers promoter 31..48 /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 54..70 /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 104..110 /note="T7 phage, gene 10 translational enhancer sequence - ribosome-binding site" /regulatory_class="ribosome_binding_site" /note="Derived using Geneious Prime 2020.1.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="T7 transl en RBS" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; T7 transl en RBS" /modified_by="BarthelS" /label="RBS" CDS 118..1908 /created_by="BarthelS" /modified_by="BarthelS" /label="10xHis-phi29 DNAP 'PovSak'" misc_feature 124..153 /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 166..177 /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 202..204 /created_by="IslomjonK" /modified_by="BarthelS" /label="M8R" unsure 331..333 /created_by="IslomjonK" /modified_by="BarthelS" /label="V51A" unsure 469..471 /created_by="IslomjonK" /modified_by="BarthelS" /label="M97T" unsure 769..771 /created_by="IslomjonK" /modified_by="BarthelS" /label="G197D" unsure 841..843 /created_by="IslomjonK" /modified_by="BarthelS" /label="E221K" unsure 1843..1845 /created_by="BarthelS" /modified_by="BarthelS" /label="K555T" unsure 1888..1890 /created_by="BarthelS" /modified_by="BarthelS" /label="D570N" terminator 1950..2007 /created_by="Daniel" /label="Terminator" primer_bind complement(2011..2030) /Mismatches=0 /%GC="55.0" /Hairpin_Tm="47.0" /Self_Dimer_Tm="15.9" /Tm="60.0" /Sequence="GAAGCCTGCATAACGCGAAG" /created_by="primer3" /note="Derived using Geneious Prime 2023.1.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="oSB0020_lvl0_seq_rev" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; oSB0020_lvl0_seq_r ev" /label="oSB0020_lvl0_seq_rev" rep_origin complement(2201..2789) /created_by="Daniel" /modified_by="Daniel" /label="pMB1" primer_bind 2423..2443 /Sequence="GGTGTAGGTCGTTCGCTCCAA" /Tm="62.4" /%GC="57.1" /Hairpin_Tm="None" /Self_Dimer_Tm="8.3" /created_by="Hoffmann-Becking" /note="Derived using Geneious Prime 2023.1.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="pSB0112_0_04_phi29 DNAP (p2)" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; oMHB010_qPCR_fw_pMB1_rep_origin_E (pSB0112_0_0 4_phi29 DNAP (p2))" /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; oMHB010_qPCR_fw_pMB1_rep_origin_E (pSB0112_0_0 4_phi29 DNAP (p2))" /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; oMHB010_qPCR_fw_pMB1_rep_origin_E (pSB0112_0_0 4_phi29 DNAP (p2) (digested by EcoRI))" /label="oMHB010_qPCR_fw_pMB1_rep_origin_E" primer_bind complement(2559..2582) /Sequence="CCTCGCTCTGCTAATCCTGTTACC" /Tm="62.8" /%GC="54.2" /Hairpin_Tm="None" /Self_Dimer_Tm="None" /created_by="BarthelS" /note="Derived using Geneious Prime 2023.1.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="oMHB011_qPCR_rev_pMB1_E" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; oMHB011_qPCR_rev_pM B1_E" /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; oMHB011_qPCR_rev_pM B1_E" /label="oMHB011_qPCR_rev_pMB1_E" terminator 2972..3066 /created_by="Daniel" /label="Terminator" CDS complement(3087..3746) /created_by="Daniel" /label="cat" terminator 3930..3973 /created_by="Daniel" /label="Terminator" regulatory 4052..4073 /note="catabolite activator protein binding site" /regulatory_class="other" /note="Derived using Geneious Prime 2020.1.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="CAP BS" /Transferred_Similarity="95.45%" /Primary_Match="95.45%; Adjusted-Interval-0:0/0; CAP BS" /label="CAP BS" CDS complement(<4086..5168) /gene="lacI" /SECDrawAs="Gene" /SECStyleId=1 /modified_by="BarthelS" /label="lacI CDS" promoter complement(5169..5246) /note="lacIq promoter region" /regulatory_class="promoter" /note="Derived using Geneious Prime 2020.1.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="lacIq" /Transferred_Similarity="98.72%" /Primary_Match="98.72%; Adjusted-Interval-0:0/0; lacIq" /label="lacIq" primer_bind 5513..3 /created_by="BarthelS" /Sequence="ATGCGTCCGGCGTAG" /Tm="55.9" /%GC="66.7" /Hairpin_Tm="47.9" /Self_Dimer_Tm="17.4" /modified_by="BarthelS" /note="Derived using Geneious Prime 2023.1.2 'Annotate from Database' based on nucleotide similarity" /Transferred_From="oSB0118_pETup42bp" /Transferred_Similarity="100.00%" /Primary_Match="100.00%; Adjusted-Interval-0:0/0; oSB0118_pETup42bp< /a>" /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; oSB0118_pETup42bp< /a>" /label="oSB0118_pETup42bp" ORIGIN 1 tagaggatcg agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag 61 cggataacaa ttcccctcta gaaataattt tgtttaactt taagaaggag atataccatg 121 ggccatcatc atcatcatca tcatcatcat cacagcagcg gccatatcga aggtcgtcat 181 gggtctcgaa tgccgcgtaa acgttacagc tgcgattttg aaacgacgac gaaagttgaa 241 gattgccgtg tctgggccta tggttatatg aacatcgaag accattcaga atataaaatt 301 ggcaactcgc tggatgaatt tatggcgtgg gcgctgaaag ttcaggccga cctgtacttc 361 cacaatctga aatttgatgg tgcgttcatt atcaactggc tggaacgtaa tggctttaaa 421 tggagcgccg atggtctgcc gaacacctat aatacgatta tctctcgtac cggccaatgg 481 tatatgattg atatctgcct gggctacaaa ggtaaacgca aaattcatac cgtgatctat 541 gacagcctga aaaaactgcc gtttccggtg aagaaaattg cgaaagattt caaactgacc 601 gtcctgaaag gcgatattga ctatcacaaa gaacgtccgg ttggttacaa aatcacgccg 661 gaagaatatg cgtacattaa aaacgatatc cagattatcg cagaagctct gctgattcag 721 tttaaacaag gcctggatcg catgaccgcc ggcagtgact ccctgaaaga tttcaaagat 781 atcatcacca cgaaaaaatt taagaaagtg ttcccgaccc tgagcctggg tctggataaa 841 aaagttcgtt atgcataccg cggcggtttt acgtggctga acgaccgttt caaagaaaaa 901 gaaattggcg agggtatggt ctttgatgtg aatagtctgt atccggctca gatgtactcc 961 cgcctgctgc cgtatggcga accgatcgtt ttcgagggta aatatgtctg ggatgaagac 1021 tacccgctgc atattcagca catccgttgt gaatttgaac tgaaagaagg ctatattccg 1081 accattcaaa tcaaacgtag ccgcttctat aagggtaacg aatacctgaa aagctctggc 1141 ggtgaaatcg cagacctgtg gctgagtaac gtcgatctgg aactgatgaa agaacattac 1201 gatctgtaca acgttgaata catctccggc ctgaaattta aagccaccac gggtctgttt 1261 aaagatttca ttgacaaatg gacctacatc aaaaccacgt ctgaaggtgc aatcaaacag 1321 ctggctaaac tgatgctgaa cagcctgtat ggcaaatttg catctaatcc ggatgttacc 1381 ggtaaagtcc cgtacctgaa agaaaatggc gctctgggtt ttcgcctggg cgaagaagaa 1441 accaaagatc cggtgtatac gccgatgggt gttttcatta ccgcgtgggc acgttacacc 1501 accatcaccg ccgcacaagc gtgctatgac cgcattatct actgtgatac cgactcaatt 1561 catctgaccg gcacggaaat cccggatgtg attaaagata tcgttgaccc gaaaaaactg 1621 ggttattggg cacacgaatc gacctttaaa cgtgctaaat acctgcgcca gaaaacgtac 1681 atccaagaca tctacatgaa agaagtcgat ggcaaactgg tggaaggttc accggatgac 1741 tataccgaca ttaaattttc ggtgaaatgc gccggcatga ccgataaaat taagaaagaa 1801 gtgacgttcg aaaatttcaa agtgggtttc agtcgcaaaa tgaccccgaa accggtccaa 1861 gttccgggcg gcgttgtgct ggtcgataac accttcacga tcaaataagc tttgagacca 1921 ctcatcgcga ctagtagcgg ccgctgcagt ccggcaaaaa agggcaaggt gtcaccaccc 1981 tgcccttttt ctttaaaacc gaaaagatta cttcgcgtta tgcaggcttc ctcgctcact 2041 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 2101 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 2161 caaaagccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccacagg ctccgccccc 2221 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 2281 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 2341 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 2401 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 2461 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 2521 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 2581 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 2641 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 2701 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 2761 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 2821 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 2881 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 2941 agtaaacttg gtctgacagc tcgaggcttg gattctcacc aataaaaaac gcccggcggc 3001 aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca 3061 ggagtccaag cgagctcgat atcaaattac gccccgccct gccactcatc gcagtactgt 3121 tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga 3181 atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg 3241 ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag 3301 ggattggctg aaacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt 3361 tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg 3421 tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg 3481 tgaacactat cccatatcac cagctcaccg tctttcattg ccatacgaaa ttccggatga 3541 gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc 3601 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga 3661 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg 3721 gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac 3781 tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg 3841 tgcccgatca actcgagtgc cacctgacgt ctaagaaacc attattatca tgacattaac 3901 ctataaaaat aggcgtatca cgaggcagaa tttcagataa aaaaaatcct tagctttcgc 3961 taaggatgat ttctggaatt cgcggccgct tctagagcga tgagtctggg ttgaaggctc 4021 tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt 4081 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 4141 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc 4201 agtgagactg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag 4261 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg 4321 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg 4381 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc 4441 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac 4501 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat 4561 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc 4621 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca 4681 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga 4741 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg 4801 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg 4861 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta 4921 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc 4981 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc 5041 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc 5101 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact 5161 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga 5221 aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg 5281 cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg 5341 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac 5401 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc 5461 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc 5521 ggcg //