LOCUS       sequence-132831-        6872 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  inducible gene expression in Streptomyces.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6872)
  AUTHORS   Reuther J, Gekeler C, Tiffert Y, Wohlleben W, Muth G
  TITLE     Unique conjugation mechanism in mycelial streptomycetes: a 
            DNA-binding ATPase translocates unprocessed plasmid DNA at the 
            hyphal tip.
  JOURNAL   Mol Microbiol. 2006 Jul;61(2):436-46. doi: 
            10.1111/j.1365-2958.2006.05258.x. Epub 2006 Jun 15.
  PUBMED    16776656
REFERENCE   2  (bases 1 to 6872)
  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: 
            "10.1111/j.1365-2958.2006.05258.x"; journalName: "Mol Microbiol"; 
            date: "2006-07"; volume: "61"; issue: "2"; pages: "436-46"
COMMENT     Sequence Label: pGM190
FEATURES             Location/Qualifiers
     source          1..6872
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             815..1609
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(869..888)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1479..1498
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      1893..2481
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2382..2401
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2635..2652
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2769..2790
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2805..2810,2811..2828,2829..2835)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2843..2859
                     /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     2848..2870
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2867..2883
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2867..2883
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      complement(3062..3096)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     misc_feature    3267..3323
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      complement(3471..3519)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             5234..6676
                     /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 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 gcttcacgct gccgcaagca
      481 ctcagggcgc aagggctgct aaaggaagcg gaacacgtag aaagccagtc cgcagaaacg
      541 gtgctgaccc cggatgaatg tcagctactg ggctatctgg acaagggaaa acgcaagcgc
      601 aaagagaaag caggtagctt gcagtgggct tacatggcga tagctagact gggcggtttt
      661 atggacagca agcgaaccgg aattgccagc tggggcgccc tctggtaagg ttgggaagcc
      721 ctgcaaagta aactggatgg ctttcttgcc gccaaggatc tgatggcgca ggggatcaag
      781 atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
      841 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
      901 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
      961 caagaccgac ctgtccggtg ccctgaatga actccaagac gaggcagcgc ggctatcgtg
     1021 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     1081 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     1141 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     1201 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     1261 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     1321 actgttcgcc aggctcaagg cgcggatgcc cgacggcgag gatctcgtcg tgacccatgg
     1381 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     1441 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     1501 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     1561 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     1621 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     1681 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     1741 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaaaaggat ctaggtgaag
     1801 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     1861 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     1921 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     1981 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     2041 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2101 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2161 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2221 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2281 gagcattgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     2341 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     2401 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     2461 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     2521 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     2581 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     2641 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     2701 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     2761 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     2821 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     2881 gaccatgatt acgaattgta cgtaggcgtc agctacgatg ttccggggac tgctgatccg
     2941 gtcagcaggt ggaagaggga ctggattcca aagttctcaa tgctgcttgc tgttcttgaa
     3001 tggggggtcg ttgacgacga catggctcga ttggcgcgac aagttgctgc gattctcacc
     3061 aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt
     3121 cattactgga ccggatcggg gatctgggct gagggagccg acggcacgcg gcggctcacg
     3181 gcgtggcacg cggaacgtcc gggcttgcac ctcacgtcac gtgaggaggc agcgtggacg
     3241 gcgtcagaga agggagcgga catatgaagc ttgcatgcct gcaggtcgac tctagaggat
     3301 ccccgggtac cgagctcgaa ttccccagat ctaaagtttt gtcgtctttc cagacgttag
     3361 taaatgaatt ttctgtatga ggttttgcta aacaactttc aacagtttca gcggagtgag
     3421 aatagaaagg aacaactaaa ggaattgcga ataataattt tttcacgttg aaaatctcca
     3481 aaaaaaaagg ctccaaaagg agcctttaat tgtatcggtt tatcagcttg ctttcgaggt
     3541 gaatttctta aacagcttga taccgatagt tgcgccgaca atgacaacaa ccatcgccca
     3601 cgcataaccg atatattcgg tcgctgaggc ttgcagggag tcaaaggccg cttttgcggg
     3661 atctcgtcga aggcactaga ggcctatatg cggggatcga ccgcgcgggt cccggacggg
     3721 gaagagcggg gagctttgcc agagagcgac gacttcccct tgcgttggtg attgccggtc
     3781 agggcagcca tccgccatcg tcgcgtaggg tgtcacaccc caggaatcgc gtcactgaac
     3841 acagcagccg gtaggacgac catgactgag ttggacacca tcgcaaatcc gtccgatccc
     3901 gcggtgcagc ggatcatcga tgtcaccaag ccgtcgcgat ccaacataaa gacaacgttg
     3961 atcgaggacg tcgagcccct catgcacagc atcgcggccg gggtggagtt catcgaggtc
     4021 tacggcagcg acagcagtcc ttttccatct gagttgctgg atctgtgcgg gcggcagaac
     4081 ataccggtcc gcctcatcga ctcctcgatc gtcaaccagt tgttcaaggg ggagcggaag
     4141 gccaagacat tcggcatcgc ccgcgtccct cgcccggcca ggttcggcga tatcgcgagc
     4201 cggcgtgggg acgtcgtcgt tctcgacggg gtgaagatcg tcgggaacat cggcgcgata
     4261 gtacgcacgt cgctcgcgct cggagcgtcg gggatcatcc tggtcgacag tgacatcacc
     4321 agcatcgcgg accggcgtct ccaaagggcc agccgaggtt acgtcttctc ccttcccgtc
     4381 gttctctccg gtcgcgagga ggccatcgcc ttcattcggg acagcggtat gcagctgatg
     4441 acgctcaagg cggatggcga catttccgtg aaggaactcg gggacaatcc ggatcggctg
     4501 gccttgctgt tcggcagcga aaagggtggg ccttccgacc tgttcgagga ggcgtcttcc
     4561 gcctcggttt ccatccccat gatgagccag accgagtctc tcaacgtttc cgtttccctc
     4621 ggaatcgcgc tgcacgagag gatcgacagg aatctcgcgg ccaaccgata agcgcctctg
     4681 ttcctcggac gctcggttgc tcgacctcga ttcgtcagtg atgatcaaca ggacccagaa
     4741 atggcacgag cccgggattg gcctcccggg ctcgccgccc ccgaaccggc gcagcatgcc
     4801 gagtgaggac gtttgctctg tgacgagcgt acgagagaac agcgcgtcga caccaccgcg
     4861 ccccgccgga ccgaagtgcc actgcggcac cccgatcgtc cacatccccg ggaagcgccc
     4921 caaaatctac tgctcggacg cctgccgaaa gcgcgcaaag cgtctcattg cggaccagtc
     4981 acgcagcgta gcaagcacga ctacagcggg taatccggac gcgggcttta gtagtaaggg
     5041 aaaatctccg tccactgcag gtcagagcgc tgtaactgga acgggttcta ccccaaacga
     5101 gggtgccgcg acagccgggc gggaggaccc gcgcgacgag cggttcgccc ggcgggaccg
     5161 gcaccagacg gtgtcgctga acgaggcgtt caagggctgc gggacccggc tcactgccgg
     5221 gacggccgag ctgatgtgga agccgggcga ggcgacgtgg ggcaacacgt gccgctgcaa
     5281 caacgtccac acctgcccgt ggtgcatgag ccgcatcctg gccgtacggg gctcgaacgt
     5341 gcagctcgcc gccgacggtc tcgcggacgc cgggtacggg ctgcaccttg gcacgaacac
     5401 gttgcgccac ttcgagcgca tggcgttcgg caccgtccgc aagggaatgc ggcacggcct
     5461 ggtcgcggtc ttgcacgacg gctggaaggg cgcgtacggc tcgtccggcc gccgctggcg
     5521 cacgatgcgc gacgacttcg ggatcatcgg ctacgagcgg gccttcgagg acaccttcgg
     5581 ctggggctcc ggctggcacc tgcactggca cacgctctgg gtcacccgcg aggtgctcgg
     5641 cccggacgcc caggccgcgt tccgcgacgc actcgccgga gcctgggccg ccggggtcga
     5701 atccgcgggc ggctacaccg tcagcgagac gtgcgaccgg cccggttgct cctgtgaggg
     5761 caagggccac ggcaccgacg tgcgcccact caacggcgcg gacgccgctg acggcgatgc
     5821 gggcaagcag gcccgctacc tctacaagga cggcgacaag accaagggcg gcgtcgccaa
     5881 gatcggcctg gagctggccg gacagaactt caaggccggt cgcggcgacg accgcatggg
     5941 accgctcgac ctgggcgacg cggcggccgc cgagctgcag cggcttcgtc ggcccggccc
     6001 gttcgtcgag aagtaccgcg agcgcgagtt cggcgtcttc caggtccgca agcactaccg
     6061 gtcgcagaac ctgaaccggc tgatcaagga actcggcatc cagcaggacg tccgcaccga
     6121 ggaggagatc accgacgaca ccgagggcct ggtcgccatc gcggtcatcc cggcctacat
     6181 ctggtaccga tacatcgccc gcgtcgccgg acgtcgcctc gacctcatca aggtcgccga
     6241 gacgtacggc ctaccgggcg tccggcggct cgtcgagtcc tggggcctgg tgtggggcaa
     6301 ggacgtcctg gacccgcccg ccccggaagc gccggccgcg ccgggcgacc ttgacgctga
     6361 ccagatgagg ttcgaggtca tgtccgagga ggaagctgct ttccgcgagg cgcgccggaa
     6421 ggccaacgag gcccgcacgg aggagctggc cgcttcgctc gaccgagtgc gtcagccgaa
     6481 gaaggaggcg atccgcccga cgatcagcct tcggaagcgc ctgaagccga agccggtcac
     6541 cgtcgacgtc aagactcccc cgcccggcgc cgcgtcgccc gtgtgccgcc gctgcaaggg
     6601 caagctcgcc ccggtactcc agccctgggg ccggctcccc ggagactgcc tccgcgtcga
     6661 cacagcggtc gcgtagggcc gctgagggac cgccatgaga ccgccacccc ggacatccgg
     6721 gtggcggtct catggcggtc ccggcgcgaa agccgagggt ctgcctgccg tgaggtggcc
     6781 ggcggacctg ggttactgct ggaatcccgc gcccagtccg agcgtccgcc ggcgcttttc
     6841 acctggctcc gggtgctcgt cgatgagcac ca
//
