LOCUS       sequence-69937-e        5216 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shuttle vector E. coli/B.meg., encoding a promoter for RNA 
            polymerase of the phage SP6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5216)
  AUTHORS   Stammen S, Schuller F, Dietrich S, Gamer M, Biedendieck R, Jahn D
  TITLE     Application of Escherichia coli phage K1E DNA-dependent RNA 
            polymerase for in vitro RNA synthesis and in vivo protein production
            in Bacillus megaterium.
  JOURNAL   Appl Microbiol Biotechnol. 2010 Sep;88(2):529-39. doi: 
            10.1007/s00253-010-2732-y. Epub 2010 Jul 2.
  PUBMED    20596705
REFERENCE   2  (bases 1 to 5216)
  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/s00253-010-2732-y"; journalName: "Appl Microbiol 
            Biotechnol"; date: "2010-09"; volume: "88"; issue: "2"; pages: 
            "529-39"
COMMENT     Sequence Label: pPSP6
FEATURES             Location/Qualifiers
     source          1..5216
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        807..825
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     807..824
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     rep_origin      complement(1136..1724)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1216..1235)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1895..2686,2687..2755))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2518..2537
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2756..2860)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2928..2946
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(3024..4400)
                     /codon_start=1
                     /gene="Bacillus subtilis tetL"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MNTSYSQSNLRHNQILIWLCILSFFSVLNEMVLNVSLPDIANDFN
                     KPPASTNWVNTAFMLTFSIGTAVYGKLSDQLGIKRLLLFGIIINCFGSVIGFVGHSFFS
                     LLIMARFIQGAGAAAFPALVMVVVARYIPKENRGKAFGLIGSIVAMGEGVGPAIGGMIA
                     HYIHWSYLLLIPMITIITVPFLMKLLKKEVRIKGHFDIKGIILMSVGIVFFMLFTTSYS
                     ISFLIVSVLSFLIFVKHIRKVTDPFVDPGLGKNIPFMIGVLCGGIIFGTVAGFVSMVPY
                     MMKDVHQLSTAEIGSVIIFPGTMSVIIFGYIGGILVDRRGPLYVLNIGVTFLSVSFLTA
                     SFLLETTSWFMTIIIVFVLGGLSFTKTVISTIVSSSLKQQEAGAGMSLLNFTSFLSEGT
                     GIAIVGGLLSIPLLDQRLLPMEVDQSTYLYSNLLLLFSGIIVISWLVTLNVYKHSQRDF
                     "
     CDS             complement(join(4600..5216,1..388))
                     /codon_start=1
                     /gene="repB"
                     /product="RepB replication protein"
                     /label=repB
                     /note="from Enterococcus faecalis plasmid pAM-alpha-1"
                     /db_xref="GI:22652809"
                     /protein_id="AAN03827.1"
                     /translation="MGVSFNIMCPNSSIYSDEKSRVLVDKTKSGKVRPWREKKIANVDY
                     FELLHILEFKKAERVKDCAEILEYKQNRETGERKLYRVWFCKSRLCPMCNWRRAMKHGI
                     QSQKVVAEVIKQKPTVRWLFLTLTVKNVYDGEELNKSLSDMAQGFRRMMQYKKINKNLV
                     GFMRATEVTINNKDNSYNQHMHVLVCVEPTYFKNTENYVNQKQWIQFWKKAMKLDYDPN
                     VKVQMIRPKNKYKSDIQSAIDETAKYPVKDTDFMTDDEEKNLKRLSDLEEGLHRKRLIS
                     YGGLLKEIHKKLNLDDTEEGDLIHTDDDEKADEDGFSIIAMWNWERKNYFIKE"
ORIGIN
        1 taactgttaa tgtgagaaac aaccaacgaa ctgttggctt ttgtttaata acttcagcaa
       61 caaccttttg tgactgaatg ccatgtttca ttgctctcct ccagttgcac attggacaaa
      121 gcctggattt acaaaaccac actcgataca actttctttc gcctgtttca cgattttgtt
      181 tatactctaa tatttcagca caatctttta ctctttcagc ctttttaaat tcaagaatat
      241 gcagaagttc aaagtaatca acattagcga ttttcttttc tctccatggt ctcacttttc
      301 cactttttgt cttgtccact aaaacccttg atttttcatc tgaataaatg ctactattag
      361 gacacataat attaaaagaa acccccatct atttagttat ttgtttggtc acttataact
      421 ttaacagatg gggtttttct gtgcaaccaa ttttaagggt tttcaatact ttaaaacaca
      481 tacataccaa cacttcaacg cacctttcag caactaaaat aaaaatgacg ttatttctat
      541 atgtatcaag ataagaaaga acaagttcaa aaccatcaaa aaaagacacc ttttcaggtg
      601 ctttttttat tttataaact cattccctga tctcgacttc gttctttttt tacctctcgg
      661 ttatgagtta gttcaaattc gttcttttta ggttctaaat cgtgtttttc ttggaattgt
      721 gctgttttat cctttacctt gtctacaaac cccttaaaaa cgtttttaaa ggcttttaag
      781 ccgtctgtac gttccttaag catacgattt aggtgacact atagaataga agtatagtgc
      841 cgttcttttg agcggcctat tactcaccag tcttcacggg gagggctgga tagtaaaagg
      901 aggtgaaatg tacaatggtc caaactagtt cgaagatctc cggagctccc gggatccggt
      961 accggccgca tgccggctaa acctcgcgaa cggattcacc ggtccaagaa ttggagctaa
     1021 ttaattcttg cggagaactg tgaatgcgca aaccaaccct tggcagaaca tatccatcgc
     1081 gtccgccatc tccagcagcc gcacgcggcg catctcgggc cgcgttgctg gcgtttttcc
     1141 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1201 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1261 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1321 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     1381 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     1441 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     1501 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     1561 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     1621 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     1681 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     1741 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     1801 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     1861 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     1921 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     1981 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     2041 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     2101 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     2161 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct gcaggcatcg
     2221 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     2281 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     2341 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     2401 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     2461 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata
     2521 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     2581 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     2641 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     2701 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     2761 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     2821 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     2881 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     2941 cgaggccctt tcgtcttcaa gaattcctgt tataaaaaaa ggatcaattt tgaactctct
     3001 cccaaagttg atcccttaac gatttagaaa tccctttgag aatgtttata tacattcaag
     3061 gtaaccagcc aactaatgac aatgattcct gaaaaaagta ataacaaatt actatacaga
     3121 taagttgact gatcaacttc cataggtaac aacctttgat caagtaaggg tatggataat
     3181 aaaccaccta caattgcaat acctgttccc tctgataaaa agctggtaaa gttaagcaaa
     3241 ctcattccag caccagcttc ctgctgtttc aagctacttg aaacaattgt tgatataact
     3301 gttttggtga acgaaagccc acctaaaaca aatacgatta taattgtcat gaaccatgat
     3361 gttgtttcta aaagaaagga agcagttaaa aagctaacag aaagaaatgt aactccgatg
     3421 tttaacacgt ataaaggacc tcttctatca acaagtatcc caccaatgta gccgaaaata
     3481 atgacactca ttgttccagg gaaaataatt acacttccga tttcggcagt acttagctgg
     3541 tgaacatctt tcatcatata aggaaccata gagacaaacc ctgctactgt tccaaatata
     3601 attcccccac aaagaactcc aatcataaaa ggtatatttt tccctaatcc gggatcaaca
     3661 aaaggatctg ttactttcct gatatgtttt acaaatatca ggaatgacag cacgctaacg
     3721 ataagaaaag aaatgctata tgatgttgta aacaacataa aaaatacaat gcctacagac
     3781 attagtataa ttcctttgat atcaaaatga ccttttatcc ttacttcttt ctttaataat
     3841 ttcataagaa acggaacagt gataattgtt atcataggaa tgagtagaag ataggaccaa
     3901 tgaatataat gggctatcat tccaccaatc gctggaccga ctccttctcc catggctact
     3961 atcgatccaa taagaccaaa tgctttaccc ctattttcct ttggaatata gcgcgcaact
     4021 acaaccatta cgagtgctgg aaatgcagct gcaccagccc cttgaataaa acgagccata
     4081 ataagtaagg aaaagaaaga atggccaaca aacccaatta ccgacccgaa acaatttatt
     4141 ataattccaa ataggagtaa ccttttgatg cctaattgat cagatagctt tccatataca
     4201 gctgttccaa tggaaaaggt taacataaag gctgtgttca cccagtttgt actcgcaggt
     4261 ggtttattaa aatcatttgc aatatcaggt aatgagacgt tcaaaaccat ttcatttaat
     4321 acgctaaaaa aagataaaat gcaaagccaa attaaaattt ggttgtgtcg taaattcgat
     4381 tgtgaatagg atgtattcac atttcaccct ccaataatga gggcagacgt agtttatagg
     4441 gttaatgata cgcttccctc ttttaattga accctgttac attcattaca cttcataatt
     4501 aattcctcct aaacttgatt aaaacatttt accacatata aactaagttt taaattcagt
     4561 atttcatcac ttatacaaca atatggcccg tttgttgaac tactctttaa taaaataatt
     4621 tttccgttcc caattccaca ttgcaataat agaaaatcca tcttcatcgg ctttttcgtc
     4681 atcatctgta tgaatcaaat cgccttcttc tgtgtcatca aggtttaatt ttttatgtat
     4741 ttcttttaac aaaccaccat aggagattaa ccttttacgg tgtaaacctt cctccaaatc
     4801 agacaaacgt ttcaaattct tttcttcatc atcggtcata aaatccgtat cctttacagg
     4861 atattttgca gtttcgtcaa ttgccgattg tatatccgat ttatatttat ttttcggtcg
     4921 aatcatttga acttttacat ttggatcata gtctaatttc attgcctttt tccaaaattg
     4981 aatccattgt ttttgattca cgtagttttc tgtattctta aaataagttg gttccacaca
     5041 taccaataca tgcatgtgct gattataaga attatcttta ttatttattg tcacttccgt
     5101 tgcacgcata aaaccaacaa gatttttatt aattttttta tattgcatca ttcggcgaaa
     5161 tccttgagcc atatctgaca aactcttatt taattcttcg ccatcataaa catttt
//
