LOCUS       Pet28a-AzaG             6643 bp ds-DNA     circular     18-JUL-2024
DEFINITION  .
ACCESSION   urn.local...kd-hz7s2zw
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..1489
                     /locus_tag="AzaG"
                     /ApEinfo_label="AzaG"
                     /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="azaG"
     CDS             1511..1528
                     /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      1595..1642
                     /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      1679..2134
                     /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(2227..3042)
                     /codon_start=1
                     /gene="<i>aph(3')-Ia</i>"
                     /note="confers resistance to kanamycin"
                     /product="aminoglycoside phosphotransferase"
                     /transl_table=1
                     /Transferred_Translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDN
                     VGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRT
                     PDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFR
                     LAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIF
                     DEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFH
                     LMLDEFF*"
                     /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; KanR (<a
                     href=""urn:local:.:br-hz7ptc3"">0-pET-28a</a>)"
                     /label="KanR"
     rep_origin      3164..3752
                     /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(4182..4373)
                     /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(5182..6264)
                     /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(6265..6342)
                     /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 ccatatggga accgcacctc agcaccattc
      181 gacatccgat gatccgacgc cccacctccg gatcgtcggc atcggcgccg gcccggccaa
      241 cctcagcctg gcggccctgc tgcacggcga cgccgcgcag ccgcaccggt tcttcgaccg
      301 caaggccgac ttcacctggc acgacgacca gctcatcacc ggcgcgaccc tccaggtgtc
      361 gatcttcaag gacctggtga gcctctccga cccgaccaac cggttctcgt tcctgtccta
      421 cctccacgag cacgggcgga cctaccactt cctgaacgcc aagttcgccg aggtgccgcg
      481 caccgagttc cgcaactacc tggcctgggc cgcgcggagc aacgagaaca tcgccttcgg
      541 cgaatcggtc gagcgcgtcg acttcgacgg ggtcttccac gtcaccaccg accggcgccg
      601 cgtcacggcc gacaacctcg ccgtcggcgt cggcaccgtc ccgtgggtcc ccgacttcgt
      661 ccggccccac cggggcccca cccagttcca cgtcagcgag cacctgagcc gctcgaaggc
      721 cctgggcggc aagcgcgtcg ccgtcgtcgg cggcggccag tccggcgccg aggcgttcct
      781 ggacctgctc tcgcggcccg aatccgaacg gccccagcgc gtcgtgtgga tctcccggcg
      841 ccgcaactac ttcccgatcg acgactcgac cttcaccaac gacttcttca tgcccgacta
      901 ctccgactac ttcttccggc tgcccgaggc caagcggtcc gagctcatct cggcccacct
      961 gctcgcctcg gacggcatct cggagtccac gctccgcgcg atctaccagg agctgtaccg
     1021 gctccagttc atcgagcgcg acgagcaccg gttcgggctg ctgcccaacc ggaccgtcac
     1081 cgccgccgac agggccggag gcggcgcgtg ggagctgacc ctgcaccaca acgacgagcc
     1141 cgaccagacc cagtgcctcg cggtcgacgc ggtcgtgtgg gccaccggct tccggcccgc
     1201 cccgaagccg ttcctggaac cgctgctcga ccgcatggcg accgagcacg gcgagttccg
     1261 cgtggacgag gacttcgcgg ccgagtggga cggcccgccg gaccggagca tcttcgtgca
     1321 gaacgcggtg cgcggccagc gcggcctgcc cgacgtcaac ctgagcctca acgcctggcg
     1381 ggcccggcgc atcgccgacc gcctgcgcgg gcgccccgcc gccgacccgg cgatgtcgtt
     1441 catcgagtgg tcggccaagc acgccaggga gacgcagtgg acgctgtaga agcttgcggc
     1501 cgcactcgag caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga
     1561 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     1621 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac
     1681 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1741 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1801 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1861 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     1921 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1981 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2041 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2101 gcgaatttta acaaaatatt aacgtttaca atttcaggtg gcacttttcg gggaaatgtg
     2161 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaat
     2221 taattcttag aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt
     2281 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
     2341 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     2401 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     2461 gacgactgaa tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac
     2521 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     2581 tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg
     2641 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     2701 aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca
     2761 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     2821 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     2881 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg
     2941 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     3001 tcgcggccta gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact
     3061 gtttatgtaa gcagacagtt ttattgttca tgaccaaaat cccttaacgt gagttttcgt
     3121 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3181 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3241 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3301 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3361 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3421 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3481 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3541 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3601 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3661 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3721 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3781 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3841 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3901 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     3961 cgcatctgtg cggtatttca caccgcatat atggtgcact ctcagtacaa tctgctctga
     4021 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     4081 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     4141 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     4201 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     4261 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     4321 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     4381 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     4441 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     4501 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
     4561 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
     4621 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
     4681 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac
     4741 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc
     4801 cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat gccggcgata
     4861 atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg
     4921 gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag
     4981 cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata
     5041 aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg
     5101 actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa
     5161 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5221 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt
     5281 ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg
     5341 agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat
     5401 ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga
     5461 gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat
     5521 ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt
     5581 ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg
     5641 attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa
     5701 tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag
     5761 tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc
     5821 aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc
     5881 cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc
     5941 tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg
     6001 atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga
     6061 ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg
     6121 aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg
     6181 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
     6241 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
     6301 tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc
     6361 ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg
     6421 ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc
     6481 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
     6541 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
     6601 cggccacgat gcgtccggcg tagaggatcg agatctcgat ccc
//
