LOCUS       Pet28a-AzaC             6826 bp ds-DNA     circular     18-JUL-2024
DEFINITION  .
ACCESSION   urn.local...1x-hz7lu91
KEYWORDS    .
SOURCE      .
  ORGANISM  .
COMMENT     ApEinfo:methylated:1.
FEATURES             Location/Qualifiers
     promoter        8..26
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="Derived using Geneious Prime 2024.0.7 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="0-pET-28a"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; T7
                     promoter (<a
                     href=""urn:local:.:br-hz7ptc3"">0-pET-28a</a>)"
                     /label="T7 promoter"
     misc_binding    27..49
                     /locus_tag="LacO"
                     /ApEinfo_label="LacO"
                     /ApEinfo_fwdcolor="#6495ed"
                     /ApEinfo_revcolor="#6495ed"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="LacO"
     protein_bind    27..51
                     /bound_moiety="<i>lac</i> repressor encoded by
                     <i>lacI</i>"
                     /note="The <i>lac</i> repressor binds to the <i>lac</i>
                     operator to inhibit transcription in <i>E. coli</i>. This
                     inhibition can be relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /note="Derived using Geneious Prime 2024.0.7 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="0-pET-28a"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; lac
                     operator (<a
                     href=""urn:local:.:br-hz7ptc3"">0-pET-28a</a>)"
                     /label="lac operator"
     RBS             82..87
                     /note="ribosome binding site"
                     /note="Derived using Geneious Prime 2024.0.7 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="0-pET-28a"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; RBS (<a
                     href=""urn:local:.:br-hz7ptc3"">0-pET-28a</a>)"
                     /label="RBS"
     CDS             95..97
                     /product="start codon"
                     /transl_table=1
                     /Transferred_Translation="M"
                     /note="Derived using Geneious Prime 2024.0.7 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="0-pET-28a"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; ATG (<a
                     href=""urn:local:.:br-hz7ptc3"">0-pET-28a</a>)"
                     /label="ATG"
     CDS             107..124
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /Transferred_Translation="HHHHHH"
                     /note="Derived using Geneious Prime 2024.0.7 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="0-pET-28a"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; 6xHis
                     (<a href=""urn:local:.:br-hz7ptc3"">0-pET-28a</a>)"
                     /label="6xHis"
     CDS             134..151
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /transl_table=1
                     /Transferred_Translation="LVPRGS"
                     /note="Derived using Geneious Prime 2024.0.7 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="0-pET-28a"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; thrombin
                     site (<a href=""urn:local:.:br-hz7ptc3"">0-pET-28a</a>)"
                     /label="thrombin site"
     gene            158..1672
                     /locus_tag="AzaC"
                     /ApEinfo_label="AzaC"
                     /ApEinfo_fwdcolor="cyan"
                     /ApEinfo_revcolor="green"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /modified_by="Devon"
                     /label="azaC"
     CDS             1694..1711
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /Transferred_Translation="HHHHHH"
                     /note="Derived using Geneious Prime 2024.0.7 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="0-pET-28a"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; 6xHis
                     (<a href=""urn:local:.:br-hz7ptc3"">0-pET-28a</a>)"
                     /label="6xHis"
     terminator      1778..1825
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
                     /note="Derived using Geneious Prime 2024.0.7 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="0-pET-28a"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; T7
                     terminator (<a
                     href=""urn:local:.:br-hz7ptc3"">0-pET-28a</a>)"
                     /label="T7 terminator"
     rep_origin      1862..2317
                     /snapgene_direction="LEFT"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /note="Derived using Geneious Prime 2024.0.7 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="0-pET-28a"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; f1 ori
                     (<a href=""urn:local:.:br-hz7ptc3"">0-pET-28a</a>)"
                     /label="f1 ori"
     CDS             complement(2413..3225)
                     /locus_tag="KanR"
                     /ApEinfo_label="KanR"
                     /ApEinfo_fwdcolor="yellow"
                     /ApEinfo_revcolor="yellow"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="KanR"
     rep_origin      3347..3935
                     /snapgene_direction="LEFT"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /note="Derived using Geneious Prime 2024.0.7 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="0-pET-28a"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; ori (<a
                     href=""urn:local:.:br-hz7ptc3"">0-pET-28a</a>)"
                     /label="ori"
     CDS             complement(4365..4556)
                     /codon_start=1
                     /gene="<i>rop</i>"
                     /product="Rop protein"
                     /transl_table=1
                     /Transferred_Translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDAD
                     EQADICESLHDHADELYRSCLARFGDDGENL*"
                     /note="Derived using Geneious Prime 2024.0.7 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="0-pET-28a"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; rop (<a
                     href=""urn:local:.:br-hz7ptc3"">0-pET-28a</a>)"
                     /label="rop"
     CDS             complement(5365..6447)
                     /codon_start=1
                     /gene="<i>lacI</i>"
                     /note="The <i>lac</i> repressor binds to the <i>lac</i>
                     operator to inhibit transcription in <i>E. coli. </i>This
                     inhibition can be relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /product="<i>lac</i> repressor"
                     /transl_table=1
                     /Transferred_Translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSA
                     KTREKVEAAMAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQL
                     GASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFL
                     DVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLT
                     RNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRV
                     GADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVS
                     LVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
                     /note="Derived using Geneious Prime 2024.0.7 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="0-pET-28a"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; lacI (<a
                     href=""urn:local:.:br-hz7ptc3"">0-pET-28a</a>)"
                     /label="lacI"
     promoter        complement(6448..6525)
                     /gene="<i>lacI</i>"
                     /note="<br>"
                     /note="Derived using Geneious Prime 2024.0.7 'Annotate
                     from Database' based on nucleotide similarity"
                     /Transferred_From="0-pET-28a"
                     /Transferred_Similarity="100.00%"
                     /Primary_Match="100.00%; Adjusted-Interval-0:0/0; lacI
                     promoter (<a
                     href=""urn:local:.:br-hz7ptc3"">0-pET-28a</a>)"
                     /label="lacI promoter"
ORIGIN      
        1 gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa
       61 ataattttgt ttaactttaa gaaggagata taccatgggc agcagccatc atcatcatca
      121 tcacagcagc ggcctggtgc cgcgcggcag ccatatgacc gcatcgatcc tcaaccgcat
      181 catcgccgaa cgtcccgccc acgacggcgt cctcaccgtc gccgggctcg gccaccgccg
      241 cgccatgccg ctcgccgaac tccacgaccg cgcccgagcc gtcgccgggg ccctgcgacg
      301 aggcggcatc ggcgagggcg accgcatcgg cgtcctcgcc gccaaccgca tcgagtggat
      361 cctgctcgat ctcgccgcgc tcatgctcaa ggcccagacc gccgggttcg aacccggcaa
      421 gttcacgccg gacgccgaac tggtcgaacg gtacgggctc gaccagctct acaccgacgc
      481 cccgatcccc gacggcgcgc cgcccgcagt gcgcccgatc gccgagatcg cgcggcacgc
      541 cgacgaggac ccgcccgaga tccatgcgcc cctttgggaa ccggaggact gcaccaccgt
      601 caagttcacc tccggctcca ccggagtccc caaaggactc gaagccacgg tcgccagcat
      661 cgactcctcg ctgcgcgacg tccagggcat gttcgaccac cgccccggcg accggctcct
      721 ggtgttcctg ccgctgtcgc tgctccagca gcgctactgg ctctactcgg cgctgcgctg
      781 gggccacgac ctcgtcgtca cgacctacca gtccgtgttc gccgtgatgc ccgccgtcga
      841 gccgaccgtc gtcatggccg tccccgggtt cttcgactcc gccaggcgcc acctcgaagg
      901 acggctcgcc gacggcgtcg acgccgccaa ggccgccgcc gagctgttcg gctcccgcat
      961 ccgctacctg tggaccggct cggcccccgc cgacccggcc atgctccgct tcttcaccga
     1021 ggccggcctg cccatcttcg aaggctacgg cctcaacgag acctgcatcg ccaccaagaa
     1081 ccaccccggc gcccaccgcg aaggcagcgt cggcaaggcc ctgcccggca aggagatcct
     1141 catcgacgcc gacggcgtcg tgtgcgtccg cagcgaccac cccgtcaaca ccgcctacac
     1201 cttcgcggcg ccaggcgaga gcgagcggat gttcaagccc ggcgggctcg tccgcaccgg
     1261 cgacctcggc cacctcgacg aggacggcta cctgtacatc cacggccgca gcgacgacgt
     1321 catcgtcctc gacaacggca agaaggtcgt cgtccgcccg atcgagaccc gcttccgcga
     1381 caccggcgcc gtcgccgaat gcgtcctata ctgccgcaac cagaccgggc tcgtcgccgt
     1441 cgtctccccg catcccgggc ccgtcgaccg cgccgccgtc gcggccgcgc tcgccgcggt
     1501 caacgccggc tccgaacccg acgagcggat ctcccgcgtc atcaccgccg acgagccctt
     1561 caccatcgac aacggactcc tcacctccca gttcaaaccc atccgggggc gcatcctcga
     1621 gcgctaccgc gaccgcatcg ccgacccgaa agaaggactc catgcccgct gaaagcttgc
     1681 ggccgcactc gagcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa
     1741 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     1801 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg
     1861 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     1921 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     1981 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2041 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2101 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2161 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2221 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2281 aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt tcggggaaat
     2341 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2401 aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
     2461 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     2521 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     2581 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     2641 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc
     2701 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     2761 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
     2821 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     2881 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa
     2941 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     3001 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     3061 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     3121 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     3181 taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt
     3241 actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa cgtgagtttt
     3301 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3361 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3421 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3481 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3541 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3601 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3661 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3721 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3781 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3841 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3901 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3961 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4021 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     4081 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     4141 ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta caatctgctc
     4201 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct
     4261 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     4321 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     4381 tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat
     4441 tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat
     4501 gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat
     4561 gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg
     4621 atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt
     4681 aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag
     4741 cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag
     4801 atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg
     4861 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
     4921 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
     4981 agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc catgccggcg
     5041 ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg
     5101 agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga
     5161 aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg
     5221 ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag
     5281 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
     5341 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5401 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
     5461 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
     5521 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
     5581 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
     5641 cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
     5701 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
     5761 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
     5821 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
     5881 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
     5941 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
     6001 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
     6061 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
     6121 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
     6181 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     6241 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     6301 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     6361 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     6421 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     6481 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct
     6541 ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca
     6601 ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     6661 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     6721 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
     6781 tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatccc
//
