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; <a
                     href=""urn:local:.:1k9-c3uhtau"">T7</a>"
                     /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; <a
                     href=""urn:local:.:11e-c3uht2n"">lacO</a>"
                     /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; <a
                     href=""urn:local:.:1lr-c3uhtb6"">T7 transl en RBS</a>"
                     /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; <a
                     href=""urn:local:.:18i-c3uht5v"">polyhistidine</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:3/3; <a
                     href=""urn:local:.:18i-c3uht5v"">polyhistidine</a>"
                     /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; <a
                     href=""urn:local:.:11t-c3uht2r"">factor XA site</a>"
                     /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; <a
                     href=""urn:local:sbarthel:15k-e1a0lzy"">oSB0020_lvl0_seq_r
                     ev</a>"
                     /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 (<a
                     href=""urn:local:Hoffmann-Becking:1j-hda12jg"">pSB0112_0_0
                     4_phi29 DNAP (p2)</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oMHB010_qPCR_fw_pMB1_rep_origin_E (<a
                     href=""urn:local:Hoffmann-Becking:11-gaolmfx"">pSB0112_0_0
                     4_phi29 DNAP (p2)</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oMHB010_qPCR_fw_pMB1_rep_origin_E (<a
                     href=""urn:local:Hoffmann-Becking:12-gaolmfx"">pSB0112_0_0
                     4_phi29 DNAP (p2) (digested by EcoRI)</a>)"
                     /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; <a
                     href=""urn:local:barthels:3d-h29t7d9"">oMHB011_qPCR_rev_pM
                     B1_E</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:barthels:3d-h29t7d9"">oMHB011_qPCR_rev_pM
                     B1_E</a>"
                     /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; <a
                     href=""urn:local:.:1ps-c3uhtcr"">CAP BS</a>"
                     /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; <a
                     href=""urn:local:.:1g4-c3uht86"">lacIq</a>"
                     /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; <a
                     href=""urn:local:sbarthel:15x-e1a0lzy"">oSB0118_pETup42bp<
                     /a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:sbarthel:15x-e1a0lzy"">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
//
