LOCUS       sequence-69947-e        6616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shuttle vector E. coli/B.meg.; carrying the codon optimized sequence
            of 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             726..743
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     rep_origin      complement(884..1472)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(964..983)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1643..2434,2435..2503))
                     /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     2266..2285
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2504..2608)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2676..2694
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(2772..4148)
                     /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(4348..5352)
                     /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 gcggaagaac tagacaagtc agaagtcttc tcgagaataa tatttccttc taagtcggtt
       61 agaattccgt taagatagtc gactcctata tcaataccaa tcgagtagcc tgcattctta
      121 ttaaaaacaa gcattacagg tcttctgccg cctctagatt gccctgcccc aatttcaaaa
      181 ataaaatctt tttcaagcag tgtatttact tgagaggaga cagtagactt gtttaatcct
      241 gtaatctcag agagagttgc cctggagaca ggggagttct tcaaaatttc atctaatatt
      301 aatttttgat tcattttttt tactaaagct tgatctgcaa tttgaataat aaccactcct
      361 ttgtttatcc accgaactaa gttggtgttt tttgaagctt gaattagata tttaaaagta
      421 tcatatctaa tattataact aaattttcta aaaaaaacat tgacataaac atttaattaa
      481 tatatgatga gataaagtta gtttattgga taaacaaact aactcaatta agatagttga
      541 tggataaact tgttcactta aatcaaggag gtgaatgtac aatgtatatt aaaaaatgta
      601 ttggttctat tttattctta ttattattct gttcttctgc tttaccagct aaagctgata
      661 ctagttcgaa gatctccgga gctcccggga tccggtaccg gccgcatgcc ggcctgagag
      721 gatcgcatca ccatcaccat cactaaccgg tccaagaatt ggagccaatc aattcttgcg
      781 gagaactgtg aatgcgcaaa ccaacccttg gcagaacata tccatcgcgt ccgccatctc
      841 cagcagccgc acgcggcgca tctcgggccg cgttgctggc gtttttccat aggctccgcc
      901 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
      961 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     1021 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     1081 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     1141 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     1201 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     1261 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     1321 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     1381 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     1441 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     1501 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     1561 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     1621 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     1681 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     1741 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     1801 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     1861 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     1921 cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct
     1981 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     2041 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     2101 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     2161 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     2221 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac
     2281 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     2341 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     2401 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     2461 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     2521 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     2581 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     2641 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     2701 gtcttcaaga attcctgtta taaaaaaagg atcaattttg aactctctcc caaagttgat
     2761 cccttaacga tttagaaatc cctttgagaa tgtttatata cattcaaggt aaccagccaa
     2821 ctaatgacaa tgattcctga aaaaagtaat aacaaattac tatacagata agttgactga
     2881 tcaacttcca taggtaacaa cctttgatca agtaagggta tggataataa accacctaca
     2941 attgcaatac ctgttccctc tgataaaaag ctggtaaagt taagcaaact cattccagca
     3001 ccagcttcct gctgtttcaa gctacttgaa acaattgttg atataactgt tttggtgaac
     3061 gaaagcccac ctaaaacaaa tacgattata attgtcatga accatgatgt tgtttctaaa
     3121 agaaaggaag cagttaaaaa gctaacagaa agaaatgtaa ctccgatgtt taacacgtat
     3181 aaaggacctc ttctatcaac aagtatccca ccaatgtagc cgaaaataat gacactcatt
     3241 gttccaggga aaataattac acttccgatt tcggcagtac ttagctggtg aacatctttc
     3301 atcatataag gaaccataga gacaaaccct gctactgttc caaatataat tcccccacaa
     3361 agaactccaa tcataaaagg tatatttttc cctaatccgg gatcaacaaa aggatctgtt
     3421 actttcctga tatgttttac aaatatcagg aatgacagca cgctaacgat aagaaaagaa
     3481 atgctatatg atgttgtaaa caacataaaa aatacaatgc ctacagacat tagtataatt
     3541 cctttgatat caaaatgacc ttttatcctt acttctttct ttaataattt cataagaaac
     3601 ggaacagtga taattgttat cataggaatg agtagaagat aggaccaatg aatataatgg
     3661 gctatcattc caccaatcgc tggaccgact ccttctccca tggctactat cgatccaata
     3721 agaccaaatg ctttacccct attttccttt ggaatatagc gcgcaactac aaccattacg
     3781 agtgctggaa atgcagctgc accagcccct tgaataaaac gagccataat aagtaaggaa
     3841 aagaaagaat ggccaacaaa cccaattacc gacccgaaac aatttattat aattccaaat
     3901 aggagtaacc ttttgatgcc taattgatca gatagctttc catatacagc tgttccaatg
     3961 gaaaaggtta acataaaggc tgtgttcacc cagtttgtac tcgcaggtgg tttattaaaa
     4021 tcatttgcaa tatcaggtaa tgagacgttc aaaaccattt catttaatac gctaaaaaaa
     4081 gataaaatgc aaagccaaat taaaatttgg ttgtgtcgta aattcgattg tgaataggat
     4141 gtattcacat ttcaccctcc aataatgagg gcagacgtag tttatagggt taatgatacg
     4201 cttccctctt ttaattgaac cctgttacat tcattacact tcataattaa ttcctcctaa
     4261 acttgattaa aacattttac cacatataaa ctaagtttta aattcagtat ttcatcactt
     4321 atacaacaat atggcccgtt tgttgaacta ctctttaata aaataatttt tccgttccca
     4381 attccacatt gcaataatag aaaatccatc ttcatcggct ttttcgtcat catctgtatg
     4441 aatcaaatcg ccttcttctg tgtcatcaag gtttaatttt ttatgtattt cttttaacaa
     4501 accaccatag gagattaacc ttttacggtg taaaccttcc tccaaatcag acaaacgttt
     4561 caaattcttt tcttcatcat cggtcataaa atccgtatcc tttacaggat attttgcagt
     4621 ttcgtcaatt gccgattgta tatccgattt atatttattt ttcggtcgaa tcatttgaac
     4681 ttttacattt ggatcatagt ctaatttcat tgcctttttc caaaattgaa tccattgttt
     4741 ttgattcacg tagttttctg tattcttaaa ataagttggt tccacacata ccaatacatg
     4801 catgtgctga ttataagaat tatctttatt atttattgtc acttccgttg cacgcataaa
     4861 accaacaaga tttttattaa tttttttata ttgcatcatt cggcgaaatc cttgagccat
     4921 atctgacaaa ctcttattta attcttcgcc atcataaaca tttttaactg ttaatgtgag
     4981 aaacaaccaa cgaactgttg gcttttgttt aataacttca gcaacaacct tttgtgactg
     5041 aatgccatgt ttcattgctc tcctccagtt gcacattgga caaagcctgg atttacaaaa
     5101 ccacactcga tacaactttc tttcgcctgt ttcacgattt tgtttatact ctaatatttc
     5161 agcacaatct tttactcttt cagccttttt aaattcaaga atatgcagaa gttcaaagta
     5221 atcaacatta gcgattttct tttctctcca tggtctcact tttccacttt ttgtcttgtc
     5281 cactaaaacc cttgattttt catctgaata aatgctacta ttaggacaca taatattaaa
     5341 agaaaccccc atctatttag ttatttgttt ggtcacttat aactttaaca gatggggttt
     5401 ttctgtgcaa ccaattttaa gggttttcaa tactttaaaa cacatacata ccaacacttc
     5461 aacgcacctt tcagcaacta aaataaaaat gacgttattt ctatatgtat caagataaga
     5521 aagaacaagt tcaaaaccat caaaaaaaga caccttttca ggtgcttttt ttattttata
     5581 aactcattcc ctgatctcga cttcgttctt tttttacctc tcggttatga gttagttcaa
     5641 attcgttctt tttaggttct aaatcgtgtt tttcttggaa ttgtgctgtt ttatccttta
     5701 ccttgtctac aaacccctta aaaacgtttt taaaggcttt taagccgtct gtacgttcct
     5761 taagatcaac agtgatatag tttgctaacc tttgcgttca cttaactaac ttataggggt
     5821 aacacttaaa aaagaatcaa taacgataga aaccgctcct aaagcaggtg cattttttcc
     5881 taacgaagaa ggcaatagtt cacatttatt gtctaaatga gaatggactc tagaagaaac
     5941 ttcgttttta atcgtattta aaacaatggg atgagattca attatatgat ttctcaagat
     6001 aacagcttct atatcaaatg tattaaggat attggttaat ccaattccga tataaaagcc
     6061 aaagttttga agtgcattta acatttctac atcattttta tttgcgcgtt ccacaatctc
     6121 ttttcgagaa atattctttt cttctttaga gagcgaagcc agtaacgctt tttcagaagc
     6181 atataattcc caacagcctc gatttccaca gctgcatttg ggtccattaa aatctatcgt
     6241 catatgaccc atttccccag aaaaaccctg aacaccttta tacaattcgt tgttaataac
     6301 aagtccagtt ccaattccga tattaatact gatgtaaacg atgttttcat agttttttgt
     6361 cataccaaat actttttcac cgtatgctcc tgcattagct tcattttcaa caaaaaccgg
     6421 aacattaaac tcactctcaa ttaaaaactg caaatctttg atattccaat ttaagttagg
     6481 catgaaaata atttgctgat gacgatctac aaggcctgga acacaaattc ctattccgac
     6541 tagaccataa ggggactcag gcatatgggt tacaaaacca tgaataagtg caaataaaat
     6601 ctcttttact tcacta
//
