LOCUS       sequence-69946-e        6628 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 NprM under 
            control of the xylose inducible promoter .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6628)
  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 6628)
  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: pSPNprM-hp
FEATURES             Location/Qualifiers
     source          1..6628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(322..1113,1114..1182))
                     /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     945..964
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1183..1287)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1355..1373
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(1451..2827)
                     /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(3027..4031)
                     /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             6033..6050
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      complement(join(6191..6628,1..151))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6271..6290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
       61 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
      121 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
      181 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
      241 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
      301 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
      361 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
      421 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
      481 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
      541 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
      601 gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc
      661 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
      721 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
      781 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
      841 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
      901 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca
      961 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     1021 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     1081 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     1141 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     1201 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     1261 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     1321 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     1381 tcttcaagaa ttcctgttat aaaaaaagga tcaattttga actctctccc aaagttgatc
     1441 ccttaacgat ttagaaatcc ctttgagaat gtttatatac attcaaggta accagccaac
     1501 taatgacaat gattcctgaa aaaagtaata acaaattact atacagataa gttgactgat
     1561 caacttccat aggtaacaac ctttgatcaa gtaagggtat ggataataaa ccacctacaa
     1621 ttgcaatacc tgttccctct gataaaaagc tggtaaagtt aagcaaactc attccagcac
     1681 cagcttcctg ctgtttcaag ctacttgaaa caattgttga tataactgtt ttggtgaacg
     1741 aaagcccacc taaaacaaat acgattataa ttgtcatgaa ccatgatgtt gtttctaaaa
     1801 gaaaggaagc agttaaaaag ctaacagaaa gaaatgtaac tccgatgttt aacacgtata
     1861 aaggacctct tctatcaaca agtatcccac caatgtagcc gaaaataatg acactcattg
     1921 ttccagggaa aataattaca cttccgattt cggcagtact tagctggtga acatctttca
     1981 tcatataagg aaccatagag acaaaccctg ctactgttcc aaatataatt cccccacaaa
     2041 gaactccaat cataaaaggt atatttttcc ctaatccggg atcaacaaaa ggatctgtta
     2101 ctttcctgat atgttttaca aatatcagga atgacagcac gctaacgata agaaaagaaa
     2161 tgctatatga tgttgtaaac aacataaaaa atacaatgcc tacagacatt agtataattc
     2221 ctttgatatc aaaatgacct tttatcctta cttctttctt taataatttc ataagaaacg
     2281 gaacagtgat aattgttatc ataggaatga gtagaagata ggaccaatga atataatggg
     2341 ctatcattcc accaatcgct ggaccgactc cttctcccat ggctactatc gatccaataa
     2401 gaccaaatgc tttaccccta ttttcctttg gaatatagcg cgcaactaca accattacga
     2461 gtgctggaaa tgcagctgca ccagcccctt gaataaaacg agccataata agtaaggaaa
     2521 agaaagaatg gccaacaaac ccaattaccg acccgaaaca atttattata attccaaata
     2581 ggagtaacct tttgatgcct aattgatcag atagctttcc atatacagct gttccaatgg
     2641 aaaaggttaa cataaaggct gtgttcaccc agtttgtact cgcaggtggt ttattaaaat
     2701 catttgcaat atcaggtaat gagacgttca aaaccatttc atttaatacg ctaaaaaaag
     2761 ataaaatgca aagccaaatt aaaatttggt tgtgtcgtaa attcgattgt gaataggatg
     2821 tattcacatt tcaccctcca ataatgaggg cagacgtagt ttatagggtt aatgatacgc
     2881 ttccctcttt taattgaacc ctgttacatt cattacactt cataattaat tcctcctaaa
     2941 cttgattaaa acattttacc acatataaac taagttttaa attcagtatt tcatcactta
     3001 tacaacaata tggcccgttt gttgaactac tctttaataa aataattttt ccgttcccaa
     3061 ttccacattg caataataga aaatccatct tcatcggctt tttcgtcatc atctgtatga
     3121 atcaaatcgc cttcttctgt gtcatcaagg tttaattttt tatgtatttc ttttaacaaa
     3181 ccaccatagg agattaacct tttacggtgt aaaccttcct ccaaatcaga caaacgtttc
     3241 aaattctttt cttcatcatc ggtcataaaa tccgtatcct ttacaggata ttttgcagtt
     3301 tcgtcaattg ccgattgtat atccgattta tatttatttt tcggtcgaat catttgaact
     3361 tttacatttg gatcatagtc taatttcatt gcctttttcc aaaattgaat ccattgtttt
     3421 tgattcacgt agttttctgt attcttaaaa taagttggtt ccacacatac caatacatgc
     3481 atgtgctgat tataagaatt atctttatta tttattgtca cttccgttgc acgcataaaa
     3541 ccaacaagat ttttattaat ttttttatat tgcatcattc ggcgaaatcc ttgagccata
     3601 tctgacaaac tcttatttaa ttcttcgcca tcataaacat ttttaactgt taatgtgaga
     3661 aacaaccaac gaactgttgg cttttgttta ataacttcag caacaacctt ttgtgactga
     3721 atgccatgtt tcattgctct cctccagttg cacattggac aaagcctgga tttacaaaac
     3781 cacactcgat acaactttct ttcgcctgtt tcacgatttt gtttatactc taatatttca
     3841 gcacaatctt ttactctttc agccttttta aattcaagaa tatgcagaag ttcaaagtaa
     3901 tcaacattag cgattttctt ttctctccat ggtctcactt ttccactttt tgtcttgtcc
     3961 actaaaaccc ttgatttttc atctgaataa atgctactat taggacacat aatattaaaa
     4021 gaaaccccca tctatttagt tatttgtttg gtcacttata actttaacag atggggtttt
     4081 tctgtgcaac caattttaag ggttttcaat actttaaaac acatacatac caacacttca
     4141 acgcaccttt cagcaactaa aataaaaatg acgttatttc tatatgtatc aagataagaa
     4201 agaacaagtt caaaaccatc aaaaaaagac accttttcag gtgctttttt tattttataa
     4261 actcattccc tgatctcgac ttcgttcttt ttttacctct cggttatgag ttagttcaaa
     4321 ttcgttcttt ttaggttcta aatcgtgttt ttcttggaat tgtgctgttt tatcctttac
     4381 cttgtctaca aaccccttaa aaacgttttt aaaggctttt aagccgtctg tacgttcctt
     4441 aagatcaaca gtgatatagt ttgctaacct ttgcgttcac ttaactaact tataggggta
     4501 acacttaaaa aagaatcaat aacgatagaa accgctccta aagcaggtgc attttttcct
     4561 aacgaagaag gcaatagttc acatttattg tctaaatgag aatggactct agaagaaact
     4621 tcgtttttaa tcgtatttaa aacaatggga tgagattcaa ttatatgatt tctcaagata
     4681 acagcttcta tatcaaatgt attaaggata ttggttaatc caattccgat ataaaagcca
     4741 aagttttgaa gtgcatttaa catttctaca tcatttttat ttgcgcgttc cacaatctct
     4801 tttcgagaaa tattcttttc ttctttagag agcgaagcca gtaacgcttt ttcagaagca
     4861 tataattccc aacagcctcg atttccacag ctgcatttgg gtccattaaa atctatcgtc
     4921 atatgaccca tttccccaga aaaaccctga acacctttat acaattcgtt gttaataaca
     4981 agtccagttc caattccgat attaatactg atgtaaacga tgttttcata gttttttgtc
     5041 ataccaaata ctttttcacc gtatgctcct gcattagctt cattttcaac aaaaaccgga
     5101 acattaaact cactctcaat taaaaactgc aaatctttga tattccaatt taagttaggc
     5161 atgaaaataa tttgctgatg acgatctaca aggcctggaa cacaaattcc tattccgact
     5221 agaccataag gggactcagg catatgggtt acaaaaccat gaataagtgc aaataaaatc
     5281 tcttttactt cactagcgga agaactagac aagtcagaag tcttctcgag aataatattt
     5341 ccttctaagt cggttagaat tccgttaaga tagtcgactc ctatatcaat accaatcgag
     5401 tagcctgcat tcttattaaa aacaagcatt acaggtcttc tgccgcctct agattgccct
     5461 gccccaattt caaaaataaa atctttttca agcagtgtat ttacttgaga ggagacagta
     5521 gacttgttta atcctgtaat ctcagagaga gttgccctgg agacagggga gttcttcaaa
     5581 atttcatcta atattaattt ttgattcatt ttttttacta aagcttgatc tgcaatttga
     5641 ataataacca ctcctttgtt tatccaccga actaagttgg tgttttttga agcttgaatt
     5701 agatatttaa aagtatcata tctaatatta taactaaatt ttctaaaaaa aacattgaca
     5761 taaacattta attaatatat gatgagataa agttagttta ttggataaac aaactaactc
     5821 aattaagata gttgatggat aaacttgttc acttaaatca aggaggtgaa tgtacaatga
     5881 aaaagaaaaa acaggcttta aaggtattat tatcagttgg tatcctttct tcatcatttg
     5941 cttttgcaca tacgagcagt gccgcgacta gttcgaagat ctccggagct cccgggatcc
     6001 ggtaccggcc gcatgccggc ctgagaggat cgcatcacca tcaccatcac taaccggtcc
     6061 aagaattgga gccaatcaat tcttgcggag aactgtgaat gcgcaaacca acccttggca
     6121 gaacatatcc atcgcgtccg ccatctccag cagccgcacg cggcgcatct cgggccgcgt
     6181 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6241 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6301 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6361 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6421 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6481 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6541 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6601 agtggtggcc taactacggc tacactag
//
