LOCUS       Pet28a-AzaQ             5572 bp ds-DNA     circular     18-JUL-2024
DEFINITION  .
ACCESSION   urn.local...kl-hz7s4p6
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             79..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"
     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             90..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             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..418
                     /locus_tag="AzaACP"
                     /ApEinfo_label="AzaACP"
                     /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="azaQ"
     CDS             440..457
                     /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      524..571
                     /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      608..1063
                     /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(1156..1971)
                     /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      2093..2681
                     /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(3111..3302)
                     /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(4111..5193)
                     /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(5194..5271)
                     /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 ccatatgccc gctgacctcg acaccctggc
      181 ccgcgaacag ctcgcgcagg tcctgcgcga ccggcccgac ccggccaccc tcgacgccgg
      241 tgaggacctc gcggagacct acgggctcac ctcgctcaac aaggtcctgt tcctcacctc
      301 gctgtgccgg gccgccgacg tcggcctcga gcgcttcacc gaggacgacc tcgcccggat
      361 gcgcacgctc ggcgacgtcg tcgaggccat gcgtccccac gccgaccaga agggatgaaa
      421 gcttgcggcc gcactcgagc accaccacca ccaccactga gatccggctg ctaacaaagc
      481 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
      541 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc
      601 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
      661 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
      721 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
      781 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
      841 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
      901 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
      961 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     1021 aaatttaacg cgaattttaa caaaatatta acgtttacaa tttcaggtgg cacttttcgg
     1081 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     1141 ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     1201 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     1261 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     1321 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     1381 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
     1441 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     1501 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
     1561 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     1621 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     1681 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     1741 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     1801 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     1861 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     1921 ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct
     1981 tgtattactg tttatgtaag cagacagttt tattgttcat gaccaaaatc ccttaacgtg
     2041 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     2101 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     2161 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     2221 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     2281 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     2341 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     2401 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     2461 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     2521 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     2581 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     2641 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     2701 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     2761 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     2821 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     2881 ttctccttac gcatctgtgc ggtatttcac accgcatata tggtgcactc tcagtacaat
     2941 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     3001 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     3061 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     3121 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     3181 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     3241 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     3301 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     3361 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     3421 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     3481 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     3541 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     3601 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     3661 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     3721 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg
     3781 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
     3841 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
     3901 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
     3961 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     4021 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     4081 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4141 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4201 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     4261 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     4321 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     4381 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     4441 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     4501 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     4561 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     4621 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     4681 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     4741 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     4801 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     4861 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     4921 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     4981 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     5041 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     5101 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     5161 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     5221 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     5281 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     5341 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     5401 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     5461 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     5521 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc cc
//
