LOCUS       sequence-69945-e        6625 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shuttle vector E. coli/B.meg.; encoding the signal peptide for 
            protein secretion of the Bacillus megaterium protein Vpr under 
            control of the xylose inducible promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6625)
  AUTHORS   Stammen S, Muller BK, Korneli C, Biedendieck R, Gamer M, Franco-Lara
            E, Jahn D
  TITLE     High-yield intra- and extracellular protein production using 
            Bacillus megaterium.
  JOURNAL   Appl Environ Microbiol. 2010 Jun;76(12):4037-46. doi: 
            10.1128/AEM.00431-10. Epub 2010 Apr 30.
  PUBMED    20435764
REFERENCE   2  (bases 1 to 6625)
  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: 
            "10.1128/AEM.00431-10"; journalName: "Appl Environ Microbiol"; date:
            "2010-06"; volume: "76"; issue: "12"; pages: "4037-46"
COMMENT     Sequence Label: pSPVpr-hp
FEATURES             Location/Qualifiers
     source          1..6625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(77..868,869..937))
                     /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     700..719
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(938..1042)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1110..1128
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(1206..2582)
                     /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(2782..3786)
                     /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"
     CDS             5785..5802
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      complement(5943..6531)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6023..6042)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
       61 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
      121 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
      181 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
      241 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
      301 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
      361 ttaatagttt gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt
      421 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
      481 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
      541 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
      601 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
      661 tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca
      721 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
      781 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
      841 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
      901 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
      961 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     1021 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     1081 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc
     1141 aagaattcct gttataaaaa aaggatcaat tttgaactct ctcccaaagt tgatccctta
     1201 acgatttaga aatccctttg agaatgttta tatacattca aggtaaccag ccaactaatg
     1261 acaatgattc ctgaaaaaag taataacaaa ttactataca gataagttga ctgatcaact
     1321 tccataggta acaacctttg atcaagtaag ggtatggata ataaaccacc tacaattgca
     1381 atacctgttc cctctgataa aaagctggta aagttaagca aactcattcc agcaccagct
     1441 tcctgctgtt tcaagctact tgaaacaatt gttgatataa ctgttttggt gaacgaaagc
     1501 ccacctaaaa caaatacgat tataattgtc atgaaccatg atgttgtttc taaaagaaag
     1561 gaagcagtta aaaagctaac agaaagaaat gtaactccga tgtttaacac gtataaagga
     1621 cctcttctat caacaagtat cccaccaatg tagccgaaaa taatgacact cattgttcca
     1681 gggaaaataa ttacacttcc gatttcggca gtacttagct ggtgaacatc tttcatcata
     1741 taaggaacca tagagacaaa ccctgctact gttccaaata taattccccc acaaagaact
     1801 ccaatcataa aaggtatatt tttccctaat ccgggatcaa caaaaggatc tgttactttc
     1861 ctgatatgtt ttacaaatat caggaatgac agcacgctaa cgataagaaa agaaatgcta
     1921 tatgatgttg taaacaacat aaaaaataca atgcctacag acattagtat aattcctttg
     1981 atatcaaaat gaccttttat ccttacttct ttctttaata atttcataag aaacggaaca
     2041 gtgataattg ttatcatagg aatgagtaga agataggacc aatgaatata atgggctatc
     2101 attccaccaa tcgctggacc gactccttct cccatggcta ctatcgatcc aataagacca
     2161 aatgctttac ccctattttc ctttggaata tagcgcgcaa ctacaaccat tacgagtgct
     2221 ggaaatgcag ctgcaccagc cccttgaata aaacgagcca taataagtaa ggaaaagaaa
     2281 gaatggccaa caaacccaat taccgacccg aaacaattta ttataattcc aaataggagt
     2341 aaccttttga tgcctaattg atcagatagc tttccatata cagctgttcc aatggaaaag
     2401 gttaacataa aggctgtgtt cacccagttt gtactcgcag gtggtttatt aaaatcattt
     2461 gcaatatcag gtaatgagac gttcaaaacc atttcattta atacgctaaa aaaagataaa
     2521 atgcaaagcc aaattaaaat ttggttgtgt cgtaaattcg attgtgaata ggatgtattc
     2581 acatttcacc ctccaataat gagggcagac gtagtttata gggttaatga tacgcttccc
     2641 tcttttaatt gaaccctgtt acattcatta cacttcataa ttaattcctc ctaaacttga
     2701 ttaaaacatt ttaccacata taaactaagt tttaaattca gtatttcatc acttatacaa
     2761 caatatggcc cgtttgttga actactcttt aataaaataa tttttccgtt cccaattcca
     2821 cattgcaata atagaaaatc catcttcatc ggctttttcg tcatcatctg tatgaatcaa
     2881 atcgccttct tctgtgtcat caaggtttaa ttttttatgt atttctttta acaaaccacc
     2941 ataggagatt aaccttttac ggtgtaaacc ttcctccaaa tcagacaaac gtttcaaatt
     3001 cttttcttca tcatcggtca taaaatccgt atcctttaca ggatattttg cagtttcgtc
     3061 aattgccgat tgtatatccg atttatattt atttttcggt cgaatcattt gaacttttac
     3121 atttggatca tagtctaatt tcattgcctt tttccaaaat tgaatccatt gtttttgatt
     3181 cacgtagttt tctgtattct taaaataagt tggttccaca cataccaata catgcatgtg
     3241 ctgattataa gaattatctt tattatttat tgtcacttcc gttgcacgca taaaaccaac
     3301 aagattttta ttaatttttt tatattgcat cattcggcga aatccttgag ccatatctga
     3361 caaactctta tttaattctt cgccatcata aacattttta actgttaatg tgagaaacaa
     3421 ccaacgaact gttggctttt gtttaataac ttcagcaaca accttttgtg actgaatgcc
     3481 atgtttcatt gctctcctcc agttgcacat tggacaaagc ctggatttac aaaaccacac
     3541 tcgatacaac tttctttcgc ctgtttcacg attttgttta tactctaata tttcagcaca
     3601 atcttttact ctttcagcct ttttaaattc aagaatatgc agaagttcaa agtaatcaac
     3661 attagcgatt ttcttttctc tccatggtct cacttttcca ctttttgtct tgtccactaa
     3721 aacccttgat ttttcatctg aataaatgct actattagga cacataatat taaaagaaac
     3781 ccccatctat ttagttattt gtttggtcac ttataacttt aacagatggg gtttttctgt
     3841 gcaaccaatt ttaagggttt tcaatacttt aaaacacata cataccaaca cttcaacgca
     3901 cctttcagca actaaaataa aaatgacgtt atttctatat gtatcaagat aagaaagaac
     3961 aagttcaaaa ccatcaaaaa aagacacctt ttcaggtgct ttttttattt tataaactca
     4021 ttccctgatc tcgacttcgt tcttttttta cctctcggtt atgagttagt tcaaattcgt
     4081 tctttttagg ttctaaatcg tgtttttctt ggaattgtgc tgttttatcc tttaccttgt
     4141 ctacaaaccc cttaaaaacg tttttaaagg cttttaagcc gtctgtacgt tccttaagat
     4201 caacagtgat atagtttgct aacctttgcg ttcacttaac taacttatag gggtaacact
     4261 taaaaaagaa tcaataacga tagaaaccgc tcctaaagca ggtgcatttt ttcctaacga
     4321 agaaggcaat agttcacatt tattgtctaa atgagaatgg actctagaag aaacttcgtt
     4381 tttaatcgta tttaaaacaa tgggatgaga ttcaattata tgatttctca agataacagc
     4441 ttctatatca aatgtattaa ggatattggt taatccaatt ccgatataaa agccaaagtt
     4501 ttgaagtgca tttaacattt ctacatcatt tttatttgcg cgttccacaa tctcttttcg
     4561 agaaatattc ttttcttctt tagagagcga agccagtaac gctttttcag aagcatataa
     4621 ttcccaacag cctcgatttc cacagctgca tttgggtcca ttaaaatcta tcgtcatatg
     4681 acccatttcc ccagaaaaac cctgaacacc tttatacaat tcgttgttaa taacaagtcc
     4741 agttccaatt ccgatattaa tactgatgta aacgatgttt tcatagtttt ttgtcatacc
     4801 aaatactttt tcaccgtatg ctcctgcatt agcttcattt tcaacaaaaa ccggaacatt
     4861 aaactcactc tcaattaaaa actgcaaatc tttgatattc caatttaagt taggcatgaa
     4921 aataatttgc tgatgacgat ctacaaggcc tggaacacaa attcctattc cgactagacc
     4981 ataaggggac tcaggcatat gggttacaaa accatgaata agtgcaaata aaatctcttt
     5041 tacttcacta gcggaagaac tagacaagtc agaagtcttc tcgagaataa tatttccttc
     5101 taagtcggtt agaattccgt taagatagtc gactcctata tcaataccaa tcgagtagcc
     5161 tgcattctta ttaaaaacaa gcattacagg tcttctgccg cctctagatt gccctgcccc
     5221 aatttcaaaa ataaaatctt tttcaagcag tgtatttact tgagaggaga cagtagactt
     5281 gtttaatcct gtaatctcag agagagttgc cctggagaca ggggagttct tcaaaatttc
     5341 atctaatatt aatttttgat tcattttttt tactaaagct tgatctgcaa tttgaataat
     5401 aaccactcct ttgtttatcc accgaactaa gttggtgttt tttgaagctt gaattagata
     5461 tttaaaagta tcatatctaa tattataact aaattttcta aaaaaaacat tgacataaac
     5521 atttaattaa tatatgatga gataaagtta gtttattgga taaacaaact aactcaatta
     5581 agatagttga tggataaact tgttcactta aatcaaggag gtgaatgtac aatgaaaaaa
     5641 gtatcaattc ggtctgtatt aagtacagta gcattttcgg ttgctttatc ttcttttgca
     5701 atgggagcat ccgccaacac tagttcgaag atctccggag ctcccgggat ccggtaccgg
     5761 ccgcatgccg gcctgagagg atcgcatcac catcaccatc actaaccggt ccaagaattg
     5821 gagccaatca attcttgcgg agaactgtga atgcgcaaac caacccttgg cagaacatat
     5881 ccatcgcgtc cgccatctcc agcagccgca cgcggcgcat ctcgggccgc gttgctggcg
     5941 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6001 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6061 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6121 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6181 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6241 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6301 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6361 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     6421 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6481 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6541 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6601 gtcatgagat tatcaaaaag gatct
//
