LOCUS       sequence-252222-        5242 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  modular integrative vector for the assembly of DNA fragments and 
            expression in Streptomyces species.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5242)
  AUTHORS   Aubry C, Pernodet JL, Lautru S
  TITLE     A set of modular and integrative vectors for synthetic biology in 
            Streptomyces.
  JOURNAL   Appl Environ Microbiol. 2019 Jun 7. pii: AEM.00485-19. doi: 
            10.1128/AEM.00485-19.
  PUBMED    31175189
REFERENCE   2  (bases 1 to 5242)
  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"; journalName: "Appl 
            Environ Microbiol. 2019 Jun 7. pii: AEM.00485-19. doi: 
            10.1128/AEM.00485-19."
COMMENT     Sequence Label: pOSV811
FEATURES             Location/Qualifiers
     source          1..5242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             142..153
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      845..916
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      932..959
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2527..2549)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2538..2554
                     /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)."
     promoter        complement(join(2562..2568,2569..2585,2586..2591))
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2632..2665
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     oriT            2866..2975
                     /note="incP origin of transfer"
     CDS             complement(3373..4176)
                     /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="MSRMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3484..3503)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4094..4113
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(4429..4446)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4563..5108)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(4643..4662)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5209..5242
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
ORIGIN
        1 atgcatgcgg ccgctgctag cgttttttga tctcaatcaa taaagtcgcc tatcttttca
       61 gcaacaaaac ttgattaaca tcaattttgg tatgaccaat gcaccattca tgttattctc
      121 aatagcgaag aactactaga gaaagaggag aaatactaga tgagtgtgat cgctaaacaa
      181 atgacctaca aggtttatat gtcaggcacg gtcaatggac actactttga ggtcgaaggc
      241 gatggaaaag gtaagcccta cgagggggag cagacggtaa agctcactgt caccaagggc
      301 ggacctctgc catttgcttg ggatatttta tcaccacagt gtcagtacgg aagcatacca
      361 ttcaccaagt accctgaaga catccctgac tatgtaaagc agtcattccc ggagggctat
      421 acatgggaga ggatcatgaa ctttgaagat ggtgcagtgt gtactgtcag caatgattcc
      481 agcatccaag gcaactgttt catctaccat gtcaagttct ctggtttgaa ctttcctccc
      541 aatggacctg tcatgcagaa gaagacacag ggctgggaac ccaacactga gcgtctcttt
      601 gcacgagatg gaatgctgct aggaaacaac tttatggctc tgaagttaga aggaggcggt
      661 cactatttgt gtgaatttaa aactacttac aaggcaaaga agcctgtgaa gatgccaggg
      721 tatcactatg ttgaccgcaa actggatgta accaatcaca acaaggatta cacttcggtt
      781 gagcagtgtg aaatttccat tgcacgcaaa cctgtggtcg cctaataata ctagagccag
      841 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
      901 gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt
      961 ttatatacta gtagcggccg cttaaggtca gtcatgcggg caacttagcc gagccctacg
     1021 acaccgcccg tgggaaggtg agtgtctaac tgcgtgacaa cgccagcgca cagcggcgga
     1081 caaccgcgag cacccatgga ctggcgccgc aggtgagaag cacactggcc caaggtcgag
     1141 cgcccaccca agttgcttcg ggacgaagag gtcgtgggtt caaatcccgc caccccgaca
     1201 gagaaacacc aggtgaggca gaccgtaacg ttacggtctg cctcacctgt tttctgtgcg
     1261 tgtctatctg cgtgactatc gcgccggacc ccgcttgaag atgccgtcca tgaccacagc
     1321 gccggtctgg atgacgggcc ggatctgctt ccggtagacc tcctcagtca cggccgtacc
     1381 ggagtgtccg acgagccggg agatctcctc cagcgggacg ccgcggtcgg acagcaggga
     1441 cacgaagctg tgcctcagct ccctcggtgt ccactcgtcg gcgttgatcc cgttggcatc
     1501 cttgagcgcc tggcggaagg cgcgccggac gttagtcgcg tcgagcggct tgccaacggc
     1561 cgacgagaag accaggccgt gttcctccca cttgtcaccg gcggcgagcc gttcccagcc
     1621 ctggtcctca aagtgctgcc agaggacctc cacgcaacgc gccggcaggg cgagcgttcg
     1681 ccgagacttc cgggttttcg tgtccccacc gcgccggacc gagcgccaga cggcgatgtg
     1741 cggaggctgc ggcggctcaa cgtccggact tcccttgagg aagacgtggt cccaggtcag
     1801 cgcccgcagc tcctcggtgc gcgcaccggt cagcagggcg acgacgatgt aggcgtgcat
     1861 cgacgtgccc tcggcagcat tcagcaccgc ctcggcctgg gcgaaggtga gcgccttgga
     1921 cggccggcca ggctggccct ggggcacaga gcacagctcc accacgttgc gcttcacctt
     1981 gtcacgcgcc atggcccgct tgaccgcccg gttcaggcag gagtggaccg cctgaaggct
     2041 gcgcgtgctc agagtctgag ccttggcggc cagccagcgg tcgacgtcct ctgcgctgag
     2101 gtcacgcagc ttccgggcac ccaaacccgg tatgacgtgc ttctggctta ggtgggtgca
     2161 gttctcgacg gtgcgctggt cacggccagc gagaccgtag gcaagccagt cgttcaccgc
     2221 gtcggcgacg gtgtaccccg tgggtgcgat cgcgagaccg tcttcgtggt cacgcagaac
     2281 ctctttgagc ttgttcttgg cctccgtctt ggtcttgcca ctcccccgct tgacgatccg
     2341 cttaccgctc ggatcgaagc cgaggttcgc cgtggcgatc cagcgctgtc tcttctcgtc
     2401 ccagtggagg ccgccgtcac cccggctacg tcgcttggcc atcagatggc ctctcccaac
     2461 tggtcctgaa tgaagtcgcg tacccgggga ttcccggtta ccgagctcga attccatggt
     2521 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acattatacg agccggatga
     2581 ttaattgtca acagctcatt tcagaatatt tgccagaacc gttcctgcag ggaagttcct
     2641 attctctaga aagtatagga acttcgtcca cgacgcccgt gattttgtag ccctggccga
     2701 cggccagcag gtaggccgac aggctcatgc cggccgccgc cgccttttcc tcaatcgctc
     2761 ttcgttcgtc tggaaggcag tacaccttga taggtgggct gcccttcctg gttggcttgg
     2821 tttcatcagc catccgcttg ccctcatctg ttacgccggc ggtagccggc cagcctcgca
     2881 gagcaggatt cccgttgagc accgccaggt gcgaataagg gacagtgaag aaggaacacc
     2941 cgctcgcggg tgggcctact tcacctatcc tgcccggctg acgccgttgg atacaccaag
     3001 gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata
     3061 ccgaaaaaat cgctataatg accccgaagc agggttatgc agcggaaaag ggtaccacgt
     3121 cgcggtgagt tcaggctttt tcatatctca ttgcccccgg acgagcgtct gctccgccat
     3181 tcgccgtccg ccgtgccaat cggatcagcc gtccaaatgc gggattttcg ttagtcggag
     3241 gccaaacggc attgagcgtc agcatatcat cagcgagctg aagaaagaca atccccgatc
     3301 cgctccacgt gttgccccag caatcagcgc gaccttgccc ctccaacgtc atctcgttct
     3361 ccgctcatga gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc
     3421 gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc
     3481 agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc
     3541 acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc
     3601 gccatgggtc acgacgagat cctcgccgtc gggcatgctc gccttgagcc tggcgaacag
     3661 ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc
     3721 ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt
     3781 agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc
     3841 aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc
     3901 ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag
     3961 ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt
     4021 gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc
     4081 gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc
     4141 tgcgtgcaat ccatcttgtt caatcatgcg agacattgca ctccaccgct gatgacatca
     4201 gtcgatcata gcacgatcaa cggcactgtt gcaaatagtc ggtggtgata aacttatcat
     4261 ccccttttgc tgatggagct gcacatgaac cggatcccct agcggagtgt atactggctt
     4321 actatgttgg cactgatgag ggtgtcagtg aagtgcttca tgtggcagga gaaaaaaggc
     4381 tgcaccggtg cgtcagcaga atatgtgata caggatatat tccgcttcct cgctcactga
     4441 ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga
     4501 tttcctggaa gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg
     4561 tttttccata ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg
     4621 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg
     4681 cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg
     4741 tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat
     4801 gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4861 caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg
     4921 agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact
     4981 gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa
     5041 aaccgccctg caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac
     5101 gatctcaaga agatcatctt attaatcaga taaaatattt ctagatttca gtgcaattta
     5161 tctcttcaaa tgtagcacct gaagtcagcc ccatacgata taagttgtga agttcctatt
     5221 ctctagaaag tataggaact tc
//
