LOCUS       sequence-129702-        7002 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  inducible gene expression in Streptomyces, C-terminal His-tag 
            encoding sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7002)
  AUTHORS   Muth G
  TITLE     The pSG5-based thermosensitive vector family for genome editing and 
            gene expression in actinomycetes.
  JOURNAL   Appl Microbiol Biotechnol. 2018 Nov;102(21):9067-9080. doi: 
            10.1007/s00253-018-9334-5. Epub 2018 Sep 6.
  PUBMED    30191290
REFERENCE   2  (bases 1 to 7002)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1007/s00253-018-9334-5"; journalName: "Appl Microbiol 
            Biotechnol"; date: "2018-11"; volume: "102"; issue: "21"; pages: 
            "9067-9080"
COMMENT     Sequence Label: pGM1202
FEATURES             Location/Qualifiers
     source          1..7002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            468..577
                     /note="incP origin of transfer"
     CDS             893..1693
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                               "
                     /label=ApmR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDSFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELLAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     rep_origin      1995..2583
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2484..2503
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2737..2754
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2871..2892
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2907..2912,2913..2930,2931..2937)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2945..2961
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2950..2972
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2969..2985
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2969..2985
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      complement(3163..3197)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             3392..3409
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      complement(3601..3649)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             5365..6807
                     /codon_start=1
                     /gene="rep"
                     /product="replication initiator protein from the 
                     Streptomyces ghanaensis plasmid pSG5"
                     /label=pSG5 Rep
                     /translation="MWKPGEATWGNTCRCNNVHTCPWCMSRILAVRGSNVQLAADGLAD
                     AGYGLHLGTNTLRHFERMAFGTVRKGMRHGLVAVLHDGWKGAYGSSGRRWRTMRDDFGI
                     IGYERAFEDTFGWGSGWHLHWHTLWVTREVLGPDAQAAFRDALAGAWAAGVESAGGYTV
                     SETCDRPGCSCEGKGHGTDVRPLNGADAADGDAGKQARYLYKDGDKTKGGVAKIGLELA
                     GQNFKAGRGDDRMGPLDLGDAAAAELQRLRRPGPFVEKYREREFGVFQVRKHYRSQNLN
                     RLIKELGIQQDVRTEEEITDDTEGLVAIAVIPAYIWYRYIARVAGRRLDLIKVAETYGL
                     PGVRRLVESWGLVWGKDVLDPPAPEAPAAPGDLDADQMRFEVMSEEEAAFREARRKANE
                     ARTEELAASLDRVRQPKKEAIRPTISLRKRLKPKPVTVDVKTPPPGAASPVCRRCKGKL
                     APVLQPWGRLPGDCLRVDTAVA"
ORIGIN
        1 aggccgtcga cgacgacgcg tccgcctgcc tgcttttggc cggcgggccg ggccgcggtt
       61 gtccgccggg tcggaccggt cccttggtcg accgatgcct cggccacgac gatgctgggc
      121 ccgaagcggt acgcgaaccc gtgccgggct gcggcggatc gcggcagcag gagctccccg
      181 cccgccacgg ctggctggat ggtcgcctca cgcgcccgca ggggtgacgg cgtgggtgcg
      241 gcggtcagcc ccgtgatcgg gcgtcagacg gccgctgagg gaccgcgacc acccctccct
      301 tcccgcctgc cgtctttcac cgcccggccg gtccgcagac gccgtacggc caggccacgg
      361 aggattacgg gcagatggcg cccacccggc acccgggctg gggcaccctg gacagcacga
      421 aggtccgccc cactcaccgg ggcgggcctt cgcgttgagc tagagttccc gccagcctcg
      481 cagagcagga ttcccgttga gcaccgccag gtgcgaataa gggacagtga agaaggaaca
      541 cccgctcgcg ggtgggccta cttcacctat cctgcccggc tgacgccgtt ggatacacca
      601 aggaaagtct acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata
      661 taccgaaaaa atcgctataa tgaccccgaa gcagggttat gcagcggaaa atgcagctca
      721 cggtaactga tgccgtattt gcagtaccag cgtacggccc acagaatgat gtcacgctga
      781 aaatgccggc ctttgaatgg gttcatgtgc agctccataa gcaaaagggg atgataagtt
      841 tatcaccacc gactatttgc aacagtgccg ttgatcgtgc tatgatcgac tgatgtcatc
      901 agcggtggag tgcaatgtcg tgcaatacga atggcgaaaa gccgagctca tcggtcagct
      961 tctcaacctt ggggttaccc ccggcggtgt gctgctggtc cacagctcct tccgtagcgt
     1021 ccggcccctc gaagatgggc cacttggact gatcgaggcc ctgcgtgctg cgctgggtcc
     1081 gggagggacg ctcgtcatgc cctcgtggtc aggtctggac gacgagccgt tcgatcctgc
     1141 cacgtcgccc gttacaccgg accttggagt tgtctctgac tcattctggc gcctgccaaa
     1201 tgtaaagcgc agcgcccatc catttgcctt tgcggcagcg gggccacagg cagagcagat
     1261 catctctgat ccattgcccc tgccacctca ctcgcctgca agcccggtcg cccgtgtcca
     1321 tgaactcgat gggcaggtac ttctcctcgg cgtgggacac gatgccaaca cgacgctgca
     1381 tcttgccgag ttgttggcaa aggttcccta tggggtgccg agacactgca ccattcttca
     1441 ggatggcaag ttggtacgcg tcgattatct cgagaatgac cactgctgtg agcgctttgc
     1501 cttggcggac aggtggctca aggagaagag ccttcagaag gaaggtccag tcggtcatgc
     1561 ctttgctcgg ttgatccgct cccgcgacat tgtggcgaca gccctgggtc aactgggccg
     1621 agatccgttg atcttcctgc atccgccaga ggcgggatgc gaagaatgcg atgccgctcg
     1681 ccagtcgatt ggctgagctc ataagttcct attccgaagt tcctattcga aatgaccgac
     1741 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     1801 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     1861 atgctggagt tcttcgccca ccccaaaagg atctaggtga agatcctttt tgataatctc
     1921 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     1981 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2041 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2101 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     2161 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2221 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2281 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2341 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagcattg agaaagcgcc
     2401 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2461 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2521 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2581 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     2641 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     2701 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     2761 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     2821 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     2881 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     2941 ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat tacgaattgt
     3001 acgtaggcgt cagctacgat gttccgggga ctgctgatcc ggtcagcagg tggaagaggg
     3061 actggattcc aaagttctca atgctgcttg ctgttcttga atggggggtc gttgacgacg
     3121 acatggctcg attggcgcga caagttgctg cgattctcac caataaaaaa cgcccggcgg
     3181 caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg accggatcgg
     3241 ggatctgggc tgagggagcc gacggcacgc ggcggctcac ggcgtggcac gcggaacgtc
     3301 cgggcttgca cctcacgtca cgtgaggagg cagcgtggac ggcgtcagag aagggagcgg
     3361 acatatgaaa tctcccgccc tggccggatc ccatcatcat catcatcatt gactgcagcc
     3421 aagcttagat ccccgggtac cgagctcgaa ttccccagat ctaaagtttt gtcgtctttc
     3481 cagacgttag taaatgaatt ttctgtatga ggttttgcta aacaactttc aacagtttca
     3541 gcggagtgag aatagaaagg aacaactaaa ggaattgcga ataataattt tttcacgttg
     3601 aaaatctcca aaaaaaaagg ctccaaaagg agcctttaat tgtatcggtt tatcagcttg
     3661 ctttcgaggt gaatttctta aacagcttga taccgatagt tgcgccgaca atgacaacaa
     3721 ccatcgccca cgcataaccg atatattcgg tcgctgaggc ttgcagggag tcaaaggccg
     3781 cttttgcggg atctcgtcga aggcactaga aggcctatat gcggggatcg accgcgcggg
     3841 tcccggacgg ggaagagcgg ggagctttgc cagagagcga cgacttcccc ttgcgttggt
     3901 gattgccggt cagggcagcc atccgccatc gtcgcgtagg gtgtcacacc ccaggaatcg
     3961 cgtcactgaa cacagcagcc ggtaggacga ccatgactga gttggacacc atcgcaaatc
     4021 cgtccgatcc cgcggtgcag cggatcatcg atgtcaccaa gccgtcgcga tccaacataa
     4081 agacaacgtt gatcgaggac gtcgagcccc tcatgcacag catcgcggcc ggggtggagt
     4141 tcatcgaggt ctacggcagc gacagcagtc cttttccatc tgagttgctg gatctgtgcg
     4201 ggcggcagaa cataccggtc cgcctcatcg actcctcgat cgtcaaccag ttgttcaagg
     4261 gggagcggaa ggccaagaca ttcggcatcg cccgcgtccc tcgcccggcc aggttcggcg
     4321 atatcgcgag ccggcgtggg gacgtcgtcg ttctcgacgg ggtgaagatc gtcgggaaca
     4381 tcggcgcgat agtacgcacg tcgctcgcgc tcggagcgtc ggggatcatc ctggtcgaca
     4441 gtgacatcac cagcatcgcg gaccggcgtc tccaaagggc cagccgaggt tacgtcttct
     4501 cccttcccgt cgttctctcc ggtcgcgagg aggccatcgc cttcattcgg gacagcggta
     4561 tgcagctgat gacgctcaag gcggatggcg acatttccgt gaaggaactc ggggacaatc
     4621 cggatcggct ggccttgctg ttcggcagcg aaaagggtgg gccttccgac ctgttcgagg
     4681 aggcgtcttc cgcctcggtt tccatcccca tgatgagcca gaccgagtct ctcaacgttt
     4741 ccgtttccct cggaatcgcg ctgcacgaga ggatcgacag gaatctcgcg gccaaccgat
     4801 aagcgcctct gttcctcgga cgctcggttg ctcgacctcg attcgtcagt gatgatcaac
     4861 aggacccaga aatggcacga gcccgggatt ggcctcccgg gctcgccgcc cccgaaccgg
     4921 cgcagcatgc cgagtgagga cgtttgctct gtgacgagcg tacgagagaa cagcgcgtcg
     4981 acaccaccgc gccccgccgg accgaagtgc cactgcggca ccccgatcgt ccacatcccc
     5041 gggaagcgcc ccaaaatcta ctgctcggac gcctgccgaa agcgcgcaaa gcgtctcatt
     5101 gcggaccagt cacgcagcgt agcaagcacg actacagcgg gtaatccgga cgcgggcttt
     5161 agtagtaagg gaaaatctcc gtccactgca ggtcagagcg ctgtaactgg aacgggttct
     5221 accccaaacg agggtgccgc gacagccggg cgggaggacc cgcgcgacga gcggttcgcc
     5281 cggcgggacc ggcaccagac ggtgtcgctg aacgaggcgt tcaagggctg cgggacccgg
     5341 ctcactgccg ggacggccga gctgatgtgg aagccgggcg aggcgacgtg gggcaacacg
     5401 tgccgctgca acaacgtcca cacctgcccg tggtgcatga gccgcatcct ggccgtacgg
     5461 ggctcgaacg tgcagctcgc cgccgacggt ctcgcggacg ccgggtacgg gctgcacctt
     5521 ggcacgaaca cgttgcgcca cttcgagcgc atggcgttcg gcaccgtccg caagggaatg
     5581 cggcacggcc tggtcgcggt cttgcacgac ggctggaagg gcgcgtacgg ctcgtccggc
     5641 cgccgctggc gcacgatgcg cgacgacttc gggatcatcg gctacgagcg ggccttcgag
     5701 gacaccttcg gctggggctc cggctggcac ctgcactggc acacgctctg ggtcacccgc
     5761 gaggtgctcg gcccggacgc ccaggccgcg ttccgcgacg cactcgccgg agcctgggcc
     5821 gccggggtcg aatccgcggg cggctacacc gtcagcgaga cgtgcgaccg gcccggttgc
     5881 tcctgtgagg gcaagggcca cggcaccgac gtgcgcccac tcaacggcgc ggacgccgct
     5941 gacggcgatg cgggcaagca ggcccgctac ctctacaagg acggcgacaa gaccaagggc
     6001 ggcgtcgcca agatcggcct ggagctggcc ggacagaact tcaaggccgg tcgcggcgac
     6061 gaccgcatgg gaccgctcga cctgggcgac gcggcggccg ccgagctgca gcggcttcgt
     6121 cggcccggcc cgttcgtcga gaagtaccgc gagcgcgagt tcggcgtctt ccaggtccgc
     6181 aagcactacc ggtcgcagaa cctgaaccgg ctgatcaagg aactcggcat ccagcaggac
     6241 gtccgcaccg aggaggagat caccgacgac accgagggcc tggtcgccat cgcggtcatc
     6301 ccggcctaca tctggtaccg atacatcgcc cgcgtcgccg gacgtcgcct cgacctcatc
     6361 aaggtcgccg agacgtacgg cctaccgggc gtccggcggc tcgtcgagtc ctggggcctg
     6421 gtgtggggca aggacgtcct ggacccgccc gccccggaag cgccggccgc gccgggcgac
     6481 cttgacgctg accagatgag gttcgaggtc atgtccgagg aggaagctgc tttccgcgag
     6541 gcgcgccgga aggccaacga ggcccgcacg gaggagctgg ccgcttcgct cgaccgagtg
     6601 cgtcagccga agaaggaggc gatccgcccg acgatcagcc ttcggaagcg cctgaagccg
     6661 aagccggtca ccgtcgacgt caagactccc ccgcccggcg ccgcgtcgcc cgtgtgccgc
     6721 cgctgcaagg gcaagctcgc cccggtactc cagccctggg gccggctccc cggagactgc
     6781 ctccgcgtcg acacagcggt cgcgtagggc cgctgaggga ccgccatgag accgccaccc
     6841 cggacatccg ggtggcggtc tcatggcggt cccggcgcga aagccgaggg tctgcctgcc
     6901 gtgaggtggc cggcggacct gggttactgc tggaatcccg cgcccagtcc gagcgtccgc
     6961 cggcgctttt cacctggctc cgggtgctcg tcgatgagca cc
//
