LOCUS       pTE5430_[pSB0155_1_5C1SI-PT7-tetO-B0035-Azurite-T7term-3C1RSI]        2739 bp    DNA     circular UNA 27-OCT-2024
DEFINITION  Lvl1 MoClo vector, PT7-tetO driving Azurite, reverse TU1,
            Amp/ColE1-5C1SI-PT7-tetO-RBS0035-Azurite-T7 terminator-3C1RSI.
ACCESSION   urn.local.BarthelS.46s-idhjfyw
VERSION     urn.local.BarthelS.46s-idhjfyw
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     CDS             2..715
                     /modified_by="BarthelS"
                     /label="Azurite"
     terminator      722..768
                     //note="copied from pJBL704"
                     /created_by="sbarthel"
                     /label="T7 terminator"
     terminator      complement(796..892)
                     /ApEinfo_fwdcolor="#FF6666"
                     /ApEinfo_revcolor="#FF6666"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}"
                     /label="AmpR Terminator"
     CDS             complement(893..1750)
                     /ApEinfo_fwdcolor="#9F4240"
                     /ApEinfo_revcolor="#9F4240"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}"
                     /label="AmpR"
     primer_bind     complement(898..920)
                     /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(1751..1881)
                     /ApEinfo_fwdcolor="#FF6666"
                     /ApEinfo_revcolor="#FF6666"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}"
                     /label="AmpR Promoter"
     rep_origin      complement(1882..2645)
                     /ApEinfo_fwdcolor="#7F7F7F"
                     /ApEinfo_revcolor="#7F7F7F"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}"
                     /label="ColE1"
     primer_bind     2165..2188
                     /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>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oMHB011_qPCR_rev (<a
                     href=""urn:local:BarthelS:46r-idhjfyw"">pTE5429
                     [pSB0163_1_revTU1_PT7-lacO-RBS35-EilR-T7term]</a>)"
                     /label="oMHB011_qPCR_rev"
     primer_bind     complement(2304..2324)
                     /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>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     oMHB010_qPCR_fw (<a
                     href=""urn:local:BarthelS:46r-idhjfyw"">pTE5429
                     [pSB0163_1_revTU1_PT7-lacO-RBS35-EilR-T7term]</a>)"
                     /label="oMHB010_qPCR_fw"
     primer_bind     2571..2588
                     /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        2673..2690
                     /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          2690..2693
                     /created_by="BarthelS"
                     /modified_by="BarthelS"
                     /label="4 bp spacer"
     Operator        2694..2712
                     /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             2721..2734
                     /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>)"
                     /Alternative_Match="100.00%; Adjusted-Interval-0:0/0;
                     B0035 (<a href=""urn:local:BarthelS:46r-idhjfyw"">pTE5429
                     [pSB0163_1_revTU1_PT7-lacO-RBS35-EilR-T7term]</a>)"
                     /label="B0035"
ORIGIN      
        1 aatgtctaaa ggtgaagaac tgtttaccgg tgttgttcca attctggttg aactggatgg
       61 tgatgttaac ggtcacaaat tttctgtttc tggtgaaggc gaaggtgatg caacctacgg
      121 taaactgacc ctgaaattta tctgtaccac tggtaaactg ccagttccat ggccaactct
      181 ggttaccact ctgtctcacg gtgttcaatg tttttctcgt tacccagatc acatgaaaca
      241 gcacgatttt ttcaaaagcg caatgccaga aggttacgtt caagaacgta ccatcttctt
      301 caaagatgac ggtaactaca aaacccgtgc ggaagtgaaa tttgaaggtg ataccctggt
      361 taaccgtatc gaactgaaag gtatcgactt caaagaggat ggcaacattc tgggtcacaa
      421 actggaatac aactttaact ctcacaacat ctacatcatg gcggacaaac aaaaaaacgg
      481 catcaaagtg aacttcaaga ttcgccacaa catcgaagat ggttctgttc aactggcaga
      541 tcactaccaa caaaacaccc caattggtga tggtccagtt ctgctgccag ataaccacta
      601 cctgtctacc caatctgcac tgtctaaaga tccaaacgaa aaacgtgatc acatggtgct
      661 gctggaattt cgtaccgcag caggtattac ccacggtatg gatgaactgt acaaagctta
      721 atagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgcg cttgttggag
      781 acgagctgcg gccgcgatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
      841 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
      901 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
      961 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     1021 tgataccgcg ggacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     1081 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     1141 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     1201 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     1261 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     1321 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     1381 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     1441 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     1501 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     1561 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     1621 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     1681 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     1741 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     1801 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     1861 ttccccgaaa agtgccacct gtcatgacca aaatccctta acgtgagttt tcgttccact
     1921 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1981 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     2041 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     2101 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     2161 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     2221 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2281 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2341 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2401 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2461 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2521 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2581 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2641 accgtgcggc cgcaacacgt ctcgagtagg agtaatacga ctcactatag ggatccctat
     2701 cagtgataga gatactagag attaaagagg agaataatc
//
