LOCUS       pTE5429_[pSB0163_1_revTU1_PT7-lacO-RBS35-EilR-T7term]        2652 bp    DNA     circular UNA 27-OCT-2024
DEFINITION  Lvl1 MoClo vector, PT7-lacO driving eilR expression, reverse TU1,
            Amp/ColE1_v2-5C1SI-PT7-lacO-RBS0035-EilR-T7 terminator-3C1RSI-v2.
ACCESSION   urn.local.BarthelS.46r-idhjfyw
VERSION     urn.local.BarthelS.46r-idhjfyw
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     1383..1406
                     /created_by="Hoffmann-Becking"
                     /Sequence="CCTCGCTCTGCTAATCCTGTTACC"
                     /Tm="62.8"
                     /%GC="54.2"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /modified_by="BarthelS"
                     /note="Derived using Geneious Prime 2024.0.7 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pTE5415 [pSB0112_0_04_phi29 DNAP wt]"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0;
                     oMHB011_qPCR_rev (<a
                     href=""urn:local:BarthelS:46l-idhjfyu"">pTE5415
                     [pSB0112_0_04_phi29 DNAP wt]</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oMHB011_qPCR_rev (<a
                     href=""urn:local:BarthelS:46m-idhjfyu"">pTE5422
                     [pSB0101_1_PT7-4bp-tetO_B0035-sfGFP_1xpepper_T7term]</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oMHB011_qPCR_rev (<a
                     href=""urn:local:BarthelS:46n-idhjfyu"">pTE5425
                     [pSB0146_0_04_phi29 DNAP (p2) [D246E catalytically
                     inactive]]</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oMHB011_qPCR_rev (<a
                     href=""urn:local:BarthelS:46o-idhjfyu"">pTE5426
                     [pSB0143_PT7-tetO-B0035-phi29 DNAP-T7term]</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oMHB011_qPCR_rev (<a
                     href=""urn:local:BarthelS:46p-idhjfyu"">pTE5427
                     [pSB0161_1_TU2_PT7-tetO-RBS35-p2_dead-T7term]</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oMHB011_qPCR_rev (<a
                     href=""urn:local:BarthelS:46q-idhjfyu"">pTE5428
                     [pSB0156_1_revTU1_PT7-lacO-RBS35-TetR-T7term]</a>)"
                     /label="oMHB011_qPCR_rev"
     primer_bind     complement(1522..1542)
                     /created_by="Hoffmann-Becking"
                     /Sequence="GGTGTAGGTCGTTCGCTCCAA"
                     /Tm="62.4"
                     /%GC="57.1"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="8.3"
                     /modified_by="BarthelS"
                     /note="Derived using Geneious Prime 2024.0.7 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pTE5415 [pSB0112_0_04_phi29 DNAP wt]"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0;
                     oMHB010_qPCR_fw (<a
                     href=""urn:local:BarthelS:46l-idhjfyu"">pTE5415
                     [pSB0112_0_04_phi29 DNAP wt]</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oMHB010_qPCR_fw (<a
                     href=""urn:local:BarthelS:46m-idhjfyu"">pTE5422
                     [pSB0101_1_PT7-4bp-tetO_B0035-sfGFP_1xpepper_T7term]</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oMHB010_qPCR_fw (<a
                     href=""urn:local:BarthelS:46n-idhjfyu"">pTE5425
                     [pSB0146_0_04_phi29 DNAP (p2) [D246E catalytically
                     inactive]]</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oMHB010_qPCR_fw (<a
                     href=""urn:local:BarthelS:46o-idhjfyu"">pTE5426
                     [pSB0143_PT7-tetO-B0035-phi29 DNAP-T7term]</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oMHB010_qPCR_fw (<a
                     href=""urn:local:BarthelS:46p-idhjfyu"">pTE5427
                     [pSB0161_1_TU2_PT7-tetO-RBS35-p2_dead-T7term]</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oMHB010_qPCR_fw (<a
                     href=""urn:local:BarthelS:46q-idhjfyu"">pTE5428
                     [pSB0156_1_revTU1_PT7-lacO-RBS35-TetR-T7term]</a>)"
                     /label="oMHB010_qPCR_fw"
     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
                     /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        1914..1932
                     /kind="operator"
                     /note="Lac operon operator"
                     /note="Derived using Geneious Prime 2020.1.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /X-Transferred_From="lacO"
                     /X-Transferred_Similarity="100.00%"
                     /X-Primary_Match="100.00%; Adjusted-Interval-0:0/0; <a
                     href=""urn:local:.:11e-c3uht2n"">lacO</a>"
                     /modified_by="BarthelS"
                     /note="Derived using Geneious Prime 2024.0.7 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pTE5428
                     [pSB0156_1_revTU1_PT7-lacO-RBS35-TetR-T7term]"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; lacO (<a
                     href=""urn:local:BarthelS:46q-idhjfyu"">pTE5428
                     [pSB0156_1_revTU1_PT7-lacO-RBS35-TetR-T7term]</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; lacO
                     (<a href=""urn:local:BarthelS:46t-idhjfyx"">pTE5431
                     [pSB0160_revTU1:PT7-TetR_Spacer002_TU2:PT7-tetO-p2_wt]</a>
                     )"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; lacO
                     (<a href=""urn:local:BarthelS:46u-idhjfyx"">pTE5432
                     [pSB0162_revTU1:PT7-TetR_Spacer002_TU2:PT7-tetO-p2_wt
                     dead]</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; lacO
                     (<a href=""urn:local:BarthelS:46x-idhjfyx"">pTE5435
                     [pSB0167_revTU1:PT7-TetR_Spacer002_TU2:PT7-tetO-B0032_p2_w
                     t]</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; lacO
                     (<a href=""urn:local:BarthelS:46y-idhjfyx"">pTE5436
                     [pSB0168_revTU1:PT7-TetR_Spacer002_TU2:PT7-tetO-B0034-p2_w
                     t]</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0; lacO
                     (<a href=""urn:local:BarthelS:46z-idhjfyy"">pTE5437
                     [pSB0169_revTU1:PT7-TetR_Spacer002_TU2:PT7-tetO-B0064-p2_w
                     t]</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/-2;
                     lacO (<a href=""urn:local:BarthelS:46n-idhjfyu"">pTE5425
                     [pSB0146_0_04_phi29 DNAP (p2) [D246E catalytically
                     inactive]]</a>)"
                     /label="lacO"
     RBS             1941..1954
                     /created_by="BarthelS"
                     /note="Derived using Geneious Prime 2024.0.7 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="pTE5422
                     [pSB0101_1_PT7-4bp-tetO_B0035-sfGFP_1xpepper_T7term]"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; B0035
                     (<a href=""urn:local:BarthelS:46m-idhjfyu"">pTE5422
                     [pSB0101_1_PT7-4bp-tetO_B0035-sfGFP_1xpepper_T7term]</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     B0035 (<a href=""urn:local:BarthelS:46o-idhjfyu"">pTE5426
                     [pSB0143_PT7-tetO-B0035-phi29 DNAP-T7term]</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     B0035 (<a href=""urn:local:BarthelS:46p-idhjfyu"">pTE5427
                     [pSB0161_1_TU2_PT7-tetO-RBS35-p2_dead-T7term]</a>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     B0035 (<a href=""urn:local:BarthelS:46q-idhjfyu"">pTE5428
                     [pSB0156_1_revTU1_PT7-lacO-RBS35-TetR-T7term]</a>)"
                     /label="B0035"
     CDS             <1961..2584
                     /created_by="BarthelS"
                     /label="eilR-TEV-8xHis"
     terminator      2591..2637
                     //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 cgagtaggag taatacgact cactataggg gaattgtgag
     1921 cggataacaa tttactagag attaaagagg agaataatca atgggctatc tgaatcgcga
     1981 agaacgtcgg gaaaccatca tgcaggccgc catgcgcgtc gcgctggatc aagggtttac
     2041 cggtatgacg gtccgcaaca tcgccaccgc cgcgggcgtc gccgcaggcc aggtccacca
     2101 ccatttcacc tcctccggcg agctaaaatc ccaggcgttt atccgcgtga tccgcgagat
     2161 gatggatctg caacgccttt cgcgcaccgc cggctggcgt gaacagctgt tttccgccct
     2221 cggcagcgag gacggacggc tggagcccta tatccgcctg tggcgccagg cgcagctgct
     2281 ggccgacagc gatccggaaa tcaaaagcgc gtacctgctg accatgaacc tgtggcatga
     2341 cgaagcggtc aggatcattc gcgcgggcca tgccgctggc gaatttaccc tgcgcgatag
     2401 cgccgaaaat atcgcctggc ggctgatttc gctggtctgt ggactggacg ggatttacgt
     2461 gctgggaatg ccggaggtag acgatgccgc attcacccgc cacctgcaac acgttattca
     2521 gcttgagtta ttttcggaga atttgtactt tcaaggacat caccatcacc atcaccatca
     2581 ctgagcttaa tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgcgc
     2641 ttgttggaga cg
//
