LOCUS       sequence-459908-        6971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  OriT containing mobilizable vector for thiostrepton induced gene 
            expression in Streptomyces.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6971)
  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 6971)
  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: pGM1190
FEATURES             Location/Qualifiers
     source          1..6971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            466..575
                     /note="incP origin of transfer"
     CDS             891..1691
                     /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      1993..2581
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2482..2501
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2735..2752
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2869..2890
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2905..2910,2911..2928,2929..2935)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2943..2959
                     /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     2948..2970
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2967..2983
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2967..2983
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      complement(3161..3195)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     misc_feature    3366..3422
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      complement(3570..3618)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             5333..6775
                     /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
                     APVLQPWGQHPGDCLRVDTAVA"
ORIGIN
        1 ggccgtcgac gacgacgcgt ccgcctgcct gcttttggcc ggcgggccgg gccgcggttg
       61 tccgccgggt cggaccggtc ccttggtcga ccgatgcctc ggccacgacg atgctgggcc
      121 cgaagcggta cgcgacccgt gccgggccgc agcggatcgc ggcagcagga gctccccgcc
      181 cgccacggct ggctggatgg tcgcctcacg cgcccgcagg ggtgacggcg tgggtgcggc
      241 ggtcagcccc gtgatcgggc gtcagacggc cgctgaggga ccgcgaccac ccctcccttc
      301 ccgcctgccg tctttcaccg cccggccggt ccgcagacgc cgtacggcca ggccacggag
      361 gattacgggc agatggcgcc cacccggcac ccgggctggg gcaccctgga cagcacgaag
      421 gtccgcccca ctcaccgggg cgggccttcg cgttgagcta gagttcccgc cagcctcgca
      481 gagcaggatt cccgttgagc accgccaggt gcgaataagg gacagtgaag aaggaacacc
      541 cgctcgcggg tgggcctact tcacctatcc tgcccggctg acgccgttgg atacaccaag
      601 gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata
      661 ccgaaaaaat cgctataatg accccgaagc agggttatgc agcggaaaat gcagctcacg
      721 gtaactgatg ccgtatttgc agtaccagcg tacggcccac agaatgatgt cacgctgaaa
      781 atgccggcct ttgaatgggt tcatgtgcag ctccataagc aaaaggggat gataagttta
      841 tcaccaccga ctatttgcaa cagtgccgtt gatcgtgcta tgatcgactg atgtcatcag
      901 cggtggagtg caatgtcgtg caatacgaat ggcgaaaagc cgagctcatc ggtcagcttc
      961 tcaaccttgg ggttaccccc ggcggtgtgc tgctggtcca cagctccttc cgtagcgtcc
     1021 ggcccctcga agatgggcca cttggactga tcgaggccct gcgtgctgcg ctgggtccgg
     1081 gagggacgct cgtcatgccc tcgtggtcag gtctggacga cgagccgttc gatcctgcca
     1141 cgtcgcccgt tacaccggac cttggagttg tctctgactc attctggcgc ctgccaaatg
     1201 taaagcgcag cgcccatcca tttgcctttg cggcagcggg gccacaggca gagcagatca
     1261 tctctgatcc attgcccctg ccacctcact cgcctgcaag cccggtcgcc cgtgtccatg
     1321 aactcgatgg gcaggtactt ctcctcggcg tgggacacga tgccaacacg acgctgcatc
     1381 ttgccgagtt gttggcaaag gttccctatg gggtgccgag acactgcacc attcttcagg
     1441 atggcaagtt ggtacgcgtc gattatctcg agaatgacca ctgctgtgag cgctttgcct
     1501 tggcggacag gtggctcaag gagaagagcc ttcagaagga aggtccagtc ggtcatgcct
     1561 ttgctcggtt gatccgctcc cgcgacattg tggcgacagc cctgggtcaa ctgggccgag
     1621 atccgttgat cttcctgcat ccgccagagg cgggatgcga agaatgcgat gccgctcgcc
     1681 agtcgattgg ctgagctcat aagttcctat tccgaagttc ctattcgaaa tgaccgacca
     1741 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     1801 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     1861 gctggagttc ttcgcccacc ccaaaaggat ctaggtgaag atcctttttg ataatctcat
     1921 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     1981 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2041 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2101 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     2161 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2221 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2281 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2341 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2401 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2461 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2521 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     2581 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     2641 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     2701 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     2761 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     2821 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     2881 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     2941 aattgtgagc ggataacaat ttcacacagg aaacagctat gacatgatta cgaattgtac
     3001 gtaggcgtca gctacgatgt tccggggact gctgatccgg tcagcaggtg gaagagggac
     3061 tggattccaa agttctcaat gctgcttgct gttcttgaat ggggggtcgt tgacgacgac
     3121 atggctcgat tggcgcgaca agttgctgcg attctcacca ataaaaaacg cccggcggca
     3181 accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggac cggatcgggg
     3241 atctgggctg agggagccga cggcacgcgg cggctcacgg cgtggcacgc ggaacgtccg
     3301 ggcttgcacc tcacgtcacg tgaggaggca gcgtggacgg cgtcagagaa gggagcggac
     3361 atatgaagct tgcatgcctg caggtcgact ctagaggatc cccgggtacc gagctcgaat
     3421 tccccagatc taaagttttg tcgtctttcc agacgttagt aaatgaattt tctgtatgag
     3481 gttttgctaa acaactttca acagtttcag cggagtgaga atagaaagga acaactaaag
     3541 gaattgcgaa taataatttt ttcacgttga aaatctccaa aaaaaaaggc tccaaaagga
     3601 gcctttaatt gtatcggttt atcagcttgc tttcgaggtg aatttcttaa acagcttgat
     3661 accgatagtt gcgccgacaa tgacaacaac catcgcccac gcataaccga tatattcggt
     3721 cgctgaggct tgcagggagt caaaggccgc ttttgcggga tctcgtcgaa ggcactagag
     3781 gcctatatgc ggggatcgac cgcgcgggtc ccggacgggg aagagcgggg agctttgcca
     3841 gagagcgacg acttcccctt gcgttggtga ttgccggtca gggcagccat ccgccatcgt
     3901 cgcgtagggt gtcacacccc aggaatcgcg tcactgaaca cagcagccgg taggacgacc
     3961 atgactgagt tggacaccat cgcaaatccg tccgatcccg cggtgcagcg gatcatcgat
     4021 gtcaccaagc cgtcgcgatc caacataaag acaacgttga tcgaggacgt cgagcccctc
     4081 atgcacagca tcgcggccgg ggtggagttc atcgaggtct acggcagcga cagcagtcct
     4141 tttccatctg agttgctgga tctgtgcggg cggcagaaca taccggtccg cctcatcgac
     4201 tcctcgatcg tcaaccagtt gttcaagggg gagcggaagg ccaagacatt cggcatcgcc
     4261 cgcgtccctc gcccggccag gttcggcgat atcgcgagcc ggcgtgggga cgtcgtcgtt
     4321 ctcgacgggg tgaagatcgt cgggaacatc ggcgcgatag tacgcacgtc gctcgcgctc
     4381 ggagcgtcgg ggatcatcct ggtcgacagt gacatcacca gcatcgcgga ccggcgtctc
     4441 caaagggcca gccgaggtta cgtcttctcc cttcccgtcg ttctctccgg tcgcgaggag
     4501 gccatcgcct tcattcggga cagcggtatg cagctgatga cgctcaaggc ggatggcgac
     4561 atttccgtga aggaactcgg ggacaatccg gatcggctgg ccttgctgtt cggcagcgaa
     4621 aagggtgggc cttccgacct gttcgaggag gcgtcttccg cctcggtttc catccccatg
     4681 atgagccaga ccgagtctct caacgtttcc gtttccctcg gaatcgcgct gcacgagagg
     4741 atcgacagga atctcgcggc caaccgataa gcgcctctgt tcctcggacg ctcggttgct
     4801 cgacctcgat tcgtcagtga tgatcaacag gacccagaaa tggcacgagc ccgggattgg
     4861 cctcccgggc tcgccgcccc cgaaccggcg cagcatgccg agtgaggacg tttgctctgt
     4921 gacgagcgta cgagagaaca gcgcgtcgac accaccgcgc cccgccggac cgaagtgcca
     4981 ctgcggcacc ccgatcgtcc acatccccgg gaagcgcccc aaaatctact gctcggacgc
     5041 ctgccgaaag cgcgcaaagc gtctcattgc ggaccagtca cgcagcgtag caagcacgac
     5101 tacagcgggt aatccggacg cgggctttag tagtaaggga aaatctccgt ccactgcagg
     5161 tcagagcgct gtaactggaa cgggttctac cccaaacgag ggtgccgcga cagccgggcg
     5221 ggaggacccg cgcgacgagc ggttcgcccg gcgggaccgg caccagacgg tgtcgctgaa
     5281 cgaggcgttc aagggctgcg ggacccggct cactgccggg acggccgagc tgatgtggaa
     5341 gccgggcgag gcgacgtggg gcaacacgtg ccgctgcaac aacgtccaca cctgcccgtg
     5401 gtgcatgagc cgcatcctgg ccgtacgggg ctcgaacgtg cagctcgccg ccgacggtct
     5461 cgcggacgcc gggtacgggc tgcaccttgg cacgaacacg ttgcgccact tcgagcgcat
     5521 ggcgttcggc accgtccgca agggaatgcg gcacggcctg gtcgcggtct tgcacgacgg
     5581 ctggaagggc gcgtacggct cgtccggccg ccgctggcgc acgatgcgcg acgacttcgg
     5641 gatcatcggc tacgagcggg ccttcgagga caccttcggc tggggctccg gctggcacct
     5701 gcactggcac acgctctggg tcacccgcga ggtgctcggc ccggacgccc aggccgcgtt
     5761 ccgcgacgca ctcgccggag cctgggccgc cggggtcgaa tccgcgggcg gctacaccgt
     5821 cagcgagacg tgcgaccggc ccggttgctc ctgtgagggc aagggccacg gcaccgacgt
     5881 gcgcccactc aacggcgcgg acgccgctga cggcgatgcg ggcaagcagg cccgctacct
     5941 ctacaaggac ggcgacaaga ccaagggcgg cgtcgccaag atcggcctgg agctggccgg
     6001 acagaacttc aaggccggtc gcggcgacga ccgcatggga ccgctcgacc tgggcgacgc
     6061 ggcggccgcc gagctgcagc ggcttcgtcg gcccggcccg ttcgtcgaga agtaccgcga
     6121 gcgcgagttc ggcgtcttcc aggtccgcaa gcactaccgg tcgcagaacc tgaaccggct
     6181 gatcaaggaa ctcggcatcc agcaggacgt ccgcaccgag gaggagatca ccgacgacac
     6241 cgagggcctg gtcgccatcg cggtcatccc ggcctacatc tggtaccgat acatcgcccg
     6301 cgtcgccgga cgtcgcctcg acctcatcaa ggtcgccgag acgtacggcc taccgggcgt
     6361 ccggcggctc gtcgagtcct ggggcctggt gtggggcaag gacgtcctgg acccgcccgc
     6421 cccggaagcg ccggccgcgc cgggcgacct tgacgctgac cagatgaggt tcgaggtcat
     6481 gtccgaggag gaagctgctt tccgcgaggc gcgccggaag gccaacgagg cccgcacgga
     6541 ggagctggcc gcttcgctcg accgagtgcg tcagccgaag aaggaggcga tccgcccgac
     6601 gatcagcctt cggaagcgcc tgaagccgaa gccggtcacc gtcgacgtca agactccccc
     6661 gcccggcgcc gcgtcgcccg tgtgccgccg ctgcaagggc aagctcgccc cggtactcca
     6721 gccctggggc cagcaccccg gagactgcct ccgcgtcgac acagcggtcg cgtagggccg
     6781 ctgagggacc gccatgagac cgccaccccg gacatccggg tggcggtctc atggcggtcc
     6841 cggcgcgaaa gccgagggtc tgcctgccgt gaggtggccg gcggacctgg gttactgctg
     6901 gaatcccgcg cccagtccga gcgtccgccg gcgcttttca cctggctccg ggtgctcgtc
     6961 gatgagcacc a
//
