LOCUS       Pet28a-AzaM             6478 bp ds-DNA     circular     18-JUL-2024
DEFINITION  .
ACCESSION   urn.local...kh-hz7s47d
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..1324
                     /locus_tag="AzaM"
                     /ApEinfo_label="AzaM"
                     /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="azaM"
     CDS             1346..1363
                     /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      1430..1477
                     /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      1514..1969
                     /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(2065..2877)
                     /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      2999..3587
                     /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(4017..4208)
                     /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(5017..6099)
                     /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(6100..6177)
                     /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 cctgccaccg ctttcgaccc
      181 cgacgtgctg ggcctgccgc tgttcgagga ccggcaccgc gacctcgccg aacgcctcga
      241 ccgctggtgc tccggccgcg actgggccgg cccggccggc cccgccgacc ggtgccgcgc
      301 gctgctggcc gagatcggcg cggccggatg gctgcgccac ctcgacaccg accgccccgc
      361 aacggaactg cggtcgctgt gcgtccggcg ccaggtcctc gcctaccacg aggacctcgc
      421 cgactacgtc tactccattc aggagctgtg cgccgccgcg gtcacccgct acggcagcgg
      481 cgagcagcgc cgccgccacc tgccgggcct cgccgccggg accggcatcg gggcgctcgc
      541 gctctccgaa ccgggcgcgg gctccgacct cgccgccgtc gcgctcgagg ccgcgcccga
      601 cggcgacggc ttcgtcctca acggcgccaa gacctggatc gcccagggcg acatcgcgga
      661 cgtgctcgtc gtgctcgccc gcaccggcga cggccccggc ccgctcgggc tcaccgcgtt
      721 cctcgtcccc ggcgacgccc ccggcctcaa ggcctcgccg atcggcgcca tcgcgccccg
      781 gtcgtgggcc gagatcgcct ttaccgactg ccgcgtcggt ccggacgcgg tgctcggcga
      841 acgcggacgc ggctacgtgg tcgccctcga cgtgctcgag cgggcccgca tgaccgtcgc
      901 cgccgccgcg gtcggcttcg ccgcccgcgc cttccgcctc gcgctcgacc acgcccgcgg
      961 ccgcagggcc ttcggaagcc gcctggccga cctccagctc gtcaagtcct cgctcgcgga
     1021 catgcgcgtc aagctgtcgg cggcgtcgct gctgaccgcc cgcgccgcct gggccgccga
     1081 cagcggcgag gactacgccg aggcctcctc gaccgcgaag gtctacgcca ccgaggccgc
     1141 cggcgagatc gtcgacgcgg ccgtccagat cttcggcgcg gccgggctcg tcgccgacag
     1201 ccccgtggag cggctgtacc gccaggtccg ctccctgcgc atctacgaag gaccgtccga
     1261 ggtgctccag atgagcatcg ccgaatccct gtgaccgacg acgaccgacc gggaggcccc
     1321 atagaagctt gcggccgcac tcgagcacca ccaccaccac cactgagatc cggctgctaa
     1381 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     1441 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     1501 attggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     1561 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     1621 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     1681 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     1741 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     1801 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     1861 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     1921 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttc aggtggcact
     1981 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2041 tatccgctca tgaattaatt cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     2101 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     2161 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     2221 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     2281 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
     2341 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     2401 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
     2461 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     2521 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
     2581 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     2641 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     2701 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     2761 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     2821 catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcataac
     2881 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgacc aaaatccctt
     2941 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3001 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3061 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3121 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     3181 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3241 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3301 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3361 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3421 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3481 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3541 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3601 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3661 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     3721 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     3781 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatatggt gcactctcag
     3841 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     3901 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3961 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     4021 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     4081 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     4141 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     4201 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     4261 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     4321 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     4381 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     4441 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     4501 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     4561 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     4621 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc
     4681 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
     4741 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
     4801 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
     4861 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
     4921 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     4981 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5041 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5101 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     5161 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     5221 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     5281 gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     5341 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     5401 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     5461 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     5521 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     5581 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     5641 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     5701 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     5761 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     5821 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     5881 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     5941 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     6001 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     6061 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     6121 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     6181 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
     6241 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
     6301 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     6361 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     6421 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatccc
//
