LOCUS       sequence-69944-e        6616 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 YngK under 
            control of the xylose inducible promoter .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6616)
  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 6616)
  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: pSPYngK-hp
FEATURES             Location/Qualifiers
     source          1..6616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(109..900,901..969))
                     /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     732..751
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(970..1074)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1142..1160
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(1238..2614)
                     /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(2814..3818)
                     /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             5808..5825
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      complement(5966..6554)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6046..6065)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
       61 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
      121 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
      181 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
      241 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
      301 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
      361 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
      421 tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
      481 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
      541 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
      601 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
      661 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
      721 gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
      781 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
      841 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
      901 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
      961 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     1021 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     1081 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     1141 aaataggcgt atcacgaggc cctttcgtct tcaagaattc ctgttataaa aaaaggatca
     1201 attttgaact ctctcccaaa gttgatccct taacgattta gaaatccctt tgagaatgtt
     1261 tatatacatt caaggtaacc agccaactaa tgacaatgat tcctgaaaaa agtaataaca
     1321 aattactata cagataagtt gactgatcaa cttccatagg taacaacctt tgatcaagta
     1381 agggtatgga taataaacca cctacaattg caatacctgt tccctctgat aaaaagctgg
     1441 taaagttaag caaactcatt ccagcaccag cttcctgctg tttcaagcta cttgaaacaa
     1501 ttgttgatat aactgttttg gtgaacgaaa gcccacctaa aacaaatacg attataattg
     1561 tcatgaacca tgatgttgtt tctaaaagaa aggaagcagt taaaaagcta acagaaagaa
     1621 atgtaactcc gatgtttaac acgtataaag gacctcttct atcaacaagt atcccaccaa
     1681 tgtagccgaa aataatgaca ctcattgttc cagggaaaat aattacactt ccgatttcgg
     1741 cagtacttag ctggtgaaca tctttcatca tataaggaac catagagaca aaccctgcta
     1801 ctgttccaaa tataattccc ccacaaagaa ctccaatcat aaaaggtata tttttcccta
     1861 atccgggatc aacaaaagga tctgttactt tcctgatatg ttttacaaat atcaggaatg
     1921 acagcacgct aacgataaga aaagaaatgc tatatgatgt tgtaaacaac ataaaaaata
     1981 caatgcctac agacattagt ataattcctt tgatatcaaa atgacctttt atccttactt
     2041 ctttctttaa taatttcata agaaacggaa cagtgataat tgttatcata ggaatgagta
     2101 gaagatagga ccaatgaata taatgggcta tcattccacc aatcgctgga ccgactcctt
     2161 ctcccatggc tactatcgat ccaataagac caaatgcttt acccctattt tcctttggaa
     2221 tatagcgcgc aactacaacc attacgagtg ctggaaatgc agctgcacca gccccttgaa
     2281 taaaacgagc cataataagt aaggaaaaga aagaatggcc aacaaaccca attaccgacc
     2341 cgaaacaatt tattataatt ccaaatagga gtaacctttt gatgcctaat tgatcagata
     2401 gctttccata tacagctgtt ccaatggaaa aggttaacat aaaggctgtg ttcacccagt
     2461 ttgtactcgc aggtggttta ttaaaatcat ttgcaatatc aggtaatgag acgttcaaaa
     2521 ccatttcatt taatacgcta aaaaaagata aaatgcaaag ccaaattaaa atttggttgt
     2581 gtcgtaaatt cgattgtgaa taggatgtat tcacatttca ccctccaata atgagggcag
     2641 acgtagttta tagggttaat gatacgcttc cctcttttaa ttgaaccctg ttacattcat
     2701 tacacttcat aattaattcc tcctaaactt gattaaaaca ttttaccaca tataaactaa
     2761 gttttaaatt cagtatttca tcacttatac aacaatatgg cccgtttgtt gaactactct
     2821 ttaataaaat aatttttccg ttcccaattc cacattgcaa taatagaaaa tccatcttca
     2881 tcggcttttt cgtcatcatc tgtatgaatc aaatcgcctt cttctgtgtc atcaaggttt
     2941 aattttttat gtatttcttt taacaaacca ccataggaga ttaacctttt acggtgtaaa
     3001 ccttcctcca aatcagacaa acgtttcaaa ttcttttctt catcatcggt cataaaatcc
     3061 gtatccttta caggatattt tgcagtttcg tcaattgccg attgtatatc cgatttatat
     3121 ttatttttcg gtcgaatcat ttgaactttt acatttggat catagtctaa tttcattgcc
     3181 tttttccaaa attgaatcca ttgtttttga ttcacgtagt tttctgtatt cttaaaataa
     3241 gttggttcca cacataccaa tacatgcatg tgctgattat aagaattatc tttattattt
     3301 attgtcactt ccgttgcacg cataaaacca acaagatttt tattaatttt tttatattgc
     3361 atcattcggc gaaatccttg agccatatct gacaaactct tatttaattc ttcgccatca
     3421 taaacatttt taactgttaa tgtgagaaac aaccaacgaa ctgttggctt ttgtttaata
     3481 acttcagcaa caaccttttg tgactgaatg ccatgtttca ttgctctcct ccagttgcac
     3541 attggacaaa gcctggattt acaaaaccac actcgataca actttctttc gcctgtttca
     3601 cgattttgtt tatactctaa tatttcagca caatctttta ctctttcagc ctttttaaat
     3661 tcaagaatat gcagaagttc aaagtaatca acattagcga ttttcttttc tctccatggt
     3721 ctcacttttc cactttttgt cttgtccact aaaacccttg atttttcatc tgaataaatg
     3781 ctactattag gacacataat attaaaagaa acccccatct atttagttat ttgtttggtc
     3841 acttataact ttaacagatg gggtttttct gtgcaaccaa ttttaagggt tttcaatact
     3901 ttaaaacaca tacataccaa cacttcaacg cacctttcag caactaaaat aaaaatgacg
     3961 ttatttctat atgtatcaag ataagaaaga acaagttcaa aaccatcaaa aaaagacacc
     4021 ttttcaggtg ctttttttat tttataaact cattccctga tctcgacttc gttctttttt
     4081 tacctctcgg ttatgagtta gttcaaattc gttcttttta ggttctaaat cgtgtttttc
     4141 ttggaattgt gctgttttat cctttacctt gtctacaaac cccttaaaaa cgtttttaaa
     4201 ggcttttaag ccgtctgtac gttccttaag atcaacagtg atatagtttg ctaacctttg
     4261 cgttcactta actaacttat aggggtaaca cttaaaaaag aatcaataac gatagaaacc
     4321 gctcctaaag caggtgcatt ttttcctaac gaagaaggca atagttcaca tttattgtct
     4381 aaatgagaat ggactctaga agaaacttcg tttttaatcg tatttaaaac aatgggatga
     4441 gattcaatta tatgatttct caagataaca gcttctatat caaatgtatt aaggatattg
     4501 gttaatccaa ttccgatata aaagccaaag ttttgaagtg catttaacat ttctacatca
     4561 tttttatttg cgcgttccac aatctctttt cgagaaatat tcttttcttc tttagagagc
     4621 gaagccagta acgctttttc agaagcatat aattcccaac agcctcgatt tccacagctg
     4681 catttgggtc cattaaaatc tatcgtcata tgacccattt ccccagaaaa accctgaaca
     4741 cctttataca attcgttgtt aataacaagt ccagttccaa ttccgatatt aatactgatg
     4801 taaacgatgt tttcatagtt ttttgtcata ccaaatactt tttcaccgta tgctcctgca
     4861 ttagcttcat tttcaacaaa aaccggaaca ttaaactcac tctcaattaa aaactgcaaa
     4921 tctttgatat tccaatttaa gttaggcatg aaaataattt gctgatgacg atctacaagg
     4981 cctggaacac aaattcctat tccgactaga ccataagggg actcaggcat atgggttaca
     5041 aaaccatgaa taagtgcaaa taaaatctct tttacttcac tagcggaaga actagacaag
     5101 tcagaagtct tctcgagaat aatatttcct tctaagtcgg ttagaattcc gttaagatag
     5161 tcgactccta tatcaatacc aatcgagtag cctgcattct tattaaaaac aagcattaca
     5221 ggtcttctgc cgcctctaga ttgccctgcc ccaatttcaa aaataaaatc tttttcaagc
     5281 agtgtattta cttgagagga gacagtagac ttgtttaatc ctgtaatctc agagagagtt
     5341 gccctggaga caggggagtt cttcaaaatt tcatctaata ttaatttttg attcattttt
     5401 tttactaaag cttgatctgc aatttgaata ataaccactc ctttgtttat ccaccgaact
     5461 aagttggtgt tttttgaagc ttgaattaga tatttaaaag tatcatatct aatattataa
     5521 ctaaattttc taaaaaaaac attgacataa acatttaatt aatatatgat gagataaagt
     5581 tagtttattg gataaacaaa ctaactcaat taagatagtt gatggataaa cttgttcact
     5641 taaatcaagg aggtgaatgt acaatgtata ttaaaaaatg tataggaagc atcttgtttt
     5701 tactgctgtt ttgttcgagc gcgttacctg ctaaagcaga tactagttcg aagatctccg
     5761 gagctcccgg gatccggtac cggccgcatg ccggcctgag aggatcgcat caccatcacc
     5821 atcactaacc ggtccaagaa ttggagccaa tcaattcttg cggagaactg tgaatgcgca
     5881 aaccaaccct tggcagaaca tatccatcgc gtccgccatc tccagcagcc gcacgcggcg
     5941 catctcgggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6001 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6061 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6121 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6181 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6241 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6301 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6361 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     6421 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6481 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6541 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6601 aaaactcacg ttaagg
//
