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; <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
                     /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; <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          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; <a
                     href=""urn:local:sbarthel:15k-e1a0lzy"">oSB0020_lvl0_seq_r
                     ev</a>"
                     /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; <a
                     href=""urn:local:.:1ps-c3uhtcr"">CAP BS</a>"
                     /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; <a
                     href=""urn:local:.:1g4-c3uht86"">lacIq</a>"
                     /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; <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 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
//
