LOCUS pTE5448_[PT7-lacO-g19lead-10xHisTag-phi29_DNAP_PovSakOk] 5525 bp DNA circular UNA 22-NOV-2024 DEFINITION Expression vector for N-terminal His-tagged phi29 DNA polymerase PovSakOk. ACCESSION urn.local.barthels.10-ihb0yg2 VERSION urn.local.barthels.10-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 /SECDrawAs="Label" /modified_by="BarthelS" /label="RBS" CDS 118..1908 /created_by="BarthelS" /modified_by="BarthelS" /label="10xHis-phi29 DNAP 'PovSakOk'" 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 1372..1374 /created_by="BarthelS" /modified_by="BarthelS" /label="D398N" unsure 1447..1449 /created_by="BarthelS" /modified_by="BarthelS" /label="D423N" 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(2202..2790) /created_by="Daniel" /modified_by="Daniel" /label="pMB1" terminator 2973..3067 /created_by="Daniel" /label="Terminator" CDS complement(3088..3747) /created_by="Daniel" /label="cat" terminator 3931..3974 /created_by="Daniel" /label="Terminator" regulatory 4053..4074 /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(<4087..5169) /gene="lacI" /SECDrawAs="Gene" /SECStyleId=1 /label="lacI CDS" promoter complement(5170..5247) /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 5514..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 gaacgttacc 1381 ggtaaagtcc cgtacctgaa agaaaatggc gctctgggtt ttcgcctggg cgaagaagaa 1441 accaaaaacc 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 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccacag gctccgcccc 2221 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 2281 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 2341 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc 2401 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 2461 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 2521 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 2581 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 2641 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 2701 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 2761 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 2821 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 2881 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 2941 gagtaaactt ggtctgacag ctcgaggctt ggattctcac caataaaaaa cgcccggcgg 3001 caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac 3061 aggagtccaa gcgagctcga tatcaaatta cgccccgccc tgccactcat cgcagtactg 3121 ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg 3181 aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac 3241 gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca 3301 gggattggct gaaacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt 3361 ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg 3421 gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg 3481 gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgaa attccggatg 3541 agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt 3601 ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg 3661 agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt 3721 ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa 3781 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac 3841 gtgcccgatc aactcgagtg ccacctgacg tctaagaaac cattattatc atgacattaa 3901 cctataaaaa taggcgtatc acgaggcaga atttcagata aaaaaaatcc ttagctttcg 3961 ctaaggatga tttctggaat tcgcggccgc ttctagagcg atgagtctgg gttgaaggct 4021 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg 4081 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 4141 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac 4201 cagtgagact ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa 4261 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg 4321 gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac 4381 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac 4441 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga 4501 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata 4561 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag 4621 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc 4681 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg 4741 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat 4801 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac 4861 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt 4921 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat 4981 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc 5041 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac 5101 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac 5161 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg 5221 aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct 5281 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg 5341 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca 5401 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt 5461 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc 5521 cggcg //