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; <a
                     href=""urn:local:sbarthel:15u-e1a0lzy"">oSB0022_lvl1_seq_r
                     ev</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:sbarthel:15u-e1a0lzy"">oSB0022_lvl1_seq_r
                     ev</a>"
                     /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; <a
                     href=""urn:local:sbarthel:15n-e1a0lzy"">oSB0021_lvl1_seq_f
                     wd</a>"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:sbarthel:15n-e1a0lzy"">oSB0021_lvl1_seq_f
                     wd</a>"
                     /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 (<a
                     href=""urn:local:sbarthel:1g-d98vhsc"">T7-lacO</a>)"
                     /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 (<a
                     href=""urn:local:BarthelS:j-dbo0u8w"">T7-tetO_2bp
                     spacer</a>)"
                     /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 (<a
                     href=""urn:local:BarthelS:j-dbo0u8w"">T7-tetO_2bp
                     spacer</a>)"
                     /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; <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    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; <a
                     href=""urn:local:.:11t-c3uht2r"">factor XA site</a>"
                     /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
//
