LOCUS       pTE5451_[PT7-tetO-B0035-10xHis-phi29_DNAP_PovSak-T7term]        3816 bp    DNA     circular UNA 22-NOV-2024
DEFINITION  Lvl1 MoClo vector, PT7-tetO driving production of 10xHis-phi29 DNAP
            PovSak, RBS: B0035, TU2-N (5C2SN-PT7-tetO-RBS0035-10xHis-phi29 DNAP
            PovSak-T7term-3C2SN-AmpR-ColE1).
ACCESSION   urn.local.barthels.51-ihb2ndy
VERSION     urn.local.barthels.51-ihb2ndy
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     terminator      complement(13..109)
                     /ApEinfo_fwdcolor="#FF6666"
                     /ApEinfo_revcolor="#FF6666"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}"
                     /label="AmpR Terminator"
     CDS             complement(110..967)
                     /ApEinfo_fwdcolor="#9F4240"
                     /ApEinfo_revcolor="#9F4240"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}"
                     /label="AmpR"
     primer_bind     complement(115..137)
                     /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(968..1098)
                     /ApEinfo_fwdcolor="#FF6666"
                     /ApEinfo_revcolor="#FF6666"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}"
                     /label="AmpR Promoter"
     rep_origin      complement(1099..1862)
                     /ApEinfo_fwdcolor="#7F7F7F"
                     /ApEinfo_revcolor="#7F7F7F"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}"
                     /label="ColE1"
     primer_bind     1788..1805
                     /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        1890..1907
                     /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          1907..1910
                     /created_by="BarthelS"
                     /modified_by="BarthelS"
                     /label="4 bp spacer"
     Operator        1911..1929
                     /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             1938..1951
                     /created_by="BarthelS"
                     /label="B0035"
     CDS             1958..3748
                     /created_by="BarthelS"
                     /modified_by="BarthelS"
                     /label="10xHis phi29 DNAP 'PovSak'"
     misc_feature    1964..1993
                     /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    2006..2017
                     /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          2042..2044
                     /created_by="IslomjonK"
                     /modified_by="BarthelS"
                     /label="M8R"
     unsure          2171..2173
                     /created_by="IslomjonK"
                     /modified_by="BarthelS"
                     /label="V51A"
     unsure          2309..2311
                     /created_by="IslomjonK"
                     /modified_by="BarthelS"
                     /label="M97T"
     unsure          2609..2611
                     /created_by="IslomjonK"
                     /modified_by="BarthelS"
                     /label="G197D"
     unsure          2681..2683
                     /created_by="IslomjonK"
                     /modified_by="BarthelS"
                     /label="E221K"
     unsure          3683..3685
                     /created_by="BarthelS"
                     /modified_by="BarthelS"
                     /label="K555T"
     unsure          3728..3730
                     /created_by="BarthelS"
                     /modified_by="BarthelS"
                     /label="D570N"
     terminator      3755..3801
                     //note="copied from pJBL704"
                     /created_by="sbarthel"
                     /label="T7 terminator"
ORIGIN      
        1 agctgcggcc gcgattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
       61 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
      121 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
      181 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
      241 taccgcggga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
      301 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
      361 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
      421 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
      481 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
      541 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
      601 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
      661 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
      721 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
      781 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
      841 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
      901 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
      961 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     1021 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     1081 cccgaaaagt gccacctgtc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     1141 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     1201 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     1261 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     1321 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     1381 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     1441 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     1501 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     1561 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     1621 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     1681 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     1741 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     1801 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     1861 gtgcggccgc aacacgtctc gtactggagt aatacgactc actataggga tccctatcag
     1921 tgatagagat actagagatt aaagaggaga ataatcaatg ggccatcatc atcatcatca
     1981 tcatcatcat cacagcagcg gccatatcga aggtcgtcat ggctctcgaa tgccgcgtaa
     2041 acgttacagc tgcgattttg aaacgacgac gaaagttgaa gattgccgtg tctgggccta
     2101 tggttatatg aacatcgaag accattcaga atataaaatt ggcaactcgc tggatgaatt
     2161 tatggcgtgg gcgctgaaag ttcaggccga cctgtacttc cacaatctga aatttgatgg
     2221 tgcgttcatt atcaactggc tggaacgtaa tggctttaaa tggagcgccg atggtctgcc
     2281 gaacacctat aatacgatta tctctcgtac cggccaatgg tatatgattg atatctgcct
     2341 gggctacaaa ggtaaacgca aaattcatac cgtgatctat gacagcctga aaaaactgcc
     2401 gtttccggtg aagaaaattg cgaaagattt caaactgacc gtcctgaaag gcgatattga
     2461 ctatcacaaa gaacgtccgg ttggttacaa aatcacgccg gaagaatatg cgtacattaa
     2521 aaacgatatc cagattatcg cagaagctct gctgattcag tttaaacaag gcctggatcg
     2581 catgaccgcc ggcagtgact ccctgaaaga tttcaaagat atcatcacca cgaaaaaatt
     2641 taagaaagtg ttcccgaccc tgagcctggg tctggataaa aaagttcgtt atgcataccg
     2701 cggcggtttt acgtggctga acgaccgttt caaagaaaaa gaaattggcg agggtatggt
     2761 ctttgatgtg aatagtctgt atccggctca gatgtactcc cgcctgctgc cgtatggcga
     2821 accgatcgtt ttcgagggta aatatgtctg ggatgaagac tacccgctgc atattcagca
     2881 catccgttgt gaatttgaac tgaaagaagg ctatattccg accattcaaa tcaaacgtag
     2941 ccgcttctat aagggtaacg aatacctgaa aagctctggc ggtgaaatcg cagacctgtg
     3001 gctgagtaac gtcgatctgg aactgatgaa agaacattac gatctgtaca acgttgaata
     3061 catctccggc ctgaaattta aagccaccac gggtctgttt aaagatttca ttgacaaatg
     3121 gacctacatc aaaaccacgt ctgaaggtgc aatcaaacag ctggctaaac tgatgctgaa
     3181 cagcctgtat ggcaaatttg catctaatcc ggatgttacc ggtaaagtcc cgtacctgaa
     3241 agaaaatggc gctctgggtt ttcgcctggg cgaagaagaa accaaagatc cggtgtatac
     3301 gccgatgggt gttttcatta ccgcgtgggc acgttacacc accatcaccg ccgcacaagc
     3361 gtgctatgac cgcattatct actgtgatac cgactcaatt catctgaccg gcacggaaat
     3421 cccggatgtg attaaagata tcgttgaccc gaaaaaactg ggttattggg cacacgaatc
     3481 gacctttaaa cgtgctaaat acctgcgcca gaaaacgtac atccaagaca tctacatgaa
     3541 agaagtcgat ggcaaactgg tggaaggttc accggatgac tataccgaca ttaaattttc
     3601 ggtgaaatgc gccggcatga ccgataaaat taagaaagaa gtgacgttcg aaaatttcaa
     3661 agtgggtttc agtcgcaaaa tgaccccgaa accggtccaa gttccgggcg gcgttgtgct
     3721 ggtcgataac accttcacga tcaaataagc ttaatagcat aaccccttgg ggcctctaaa
     3781 cgggtcttga ggggtttttt gcgctaatgg gagacg
//
