LOCUS       sequence-69953-e        7166 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shuttle vector E. coli/B.meg..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7166)
  AUTHORS   Biedendieck R, Gamer M, Jaensch L, Meyer S, Rohde M, Deckwer WD, 
            Jahn D
  TITLE     A sucrose-inducible promoter system for the intra- and extracellular
            protein production in Bacillus megaterium.
  JOURNAL   J Biotechnol. 2007 Dec 1;132(4):426-30. Epub 2007 Jul 10.
  PUBMED    17692983
REFERENCE   2  (bases 1 to 7166)
  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"; journalName: "J 
            Biotechnol."; date: "2007-12-1"; volume: "132(4)"; pages: "426-30. 
            Epub 2007 Jul 10"
COMMENT     Sequence Label: pRBBm69
FEATURES             Location/Qualifiers
     source          1..7166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(2291..2879)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2371..2390)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3050..3841,3842..3910))
                     /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     3673..3692
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3911..4015)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4083..4101
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(4179..5555)
                     /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(5755..6759)
                     /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 ttaagctgat tccagccgtg aaggaaaagc aactggttta gactcttaat attcaaaaaa
       61 tgcattatac gctctctctt tttacaaaag ggagagcgta taatgcatta aaaaagataa
      121 aaatgagtag gtaaaaagaa ggaatatttt tttctgaaaa catactttag acctttatta
      181 ttatcatcct aatctttaga aaggaaaaag aatagaacgt tttcatgtcc ggctaataaa
      241 aaaaaggtac tgatttttca agagattgat aaaaacagaa tggtcggaaa gtataaaaaa
      301 cggaatagcg ttttcaatgc tggaaagata aagtgtttaa tcatgtagaa agtatcaaac
      361 gcaacgtctg gaaatcgtgg tttgaaatgt aaaaaagccg tacatttttg ttggaaaaac
      421 ggttacataa aaaaggctct tacacgtaaa ctttggtcct agttacagct caatgaaaaa
      481 gataaaaatg agaaatagct acatttggca tgtttgactt tcgtcattta tccaagtaat
      541 cttttggttg aacggaaccg gttctgtttt cggatttttg atagctaatt gaattaataa
      601 atttatattc tttactaaca caaaaggaga aaacatatga aaatgaaacg agttgctaag
      661 catacaactg ctgctacatt ggctgcagcc ctactagtag gcgggggata tcaaacattt
      721 gctaaaggaa acgacagcaa agactttaat aatagctacg gaatttcgca tattactcgc
      781 gataatatgg tgaaaattcc acagcagcaa aacagtgatc agtttaaagt tcctgctttt
      841 gatgaatcaa ctattaaaaa cattgcttct gctaaaggaa aaaatgcatc aggaaataca
      901 attgacttag atgtatggga cagctggcca ttacaaaatg ctgatggaac agtagcgact
      961 tatcatggat atcaaattgt gtttgcccta gcaggtgatc caaaagattc gaatgacacg
     1021 tctgtttatc ttttctataa aaaagctgga gacaaatcaa ttgacagctg gaaaaacgct
     1081 ggaagagtgt ttaaagacag cgataaattc gttccgaatg acccgcatct taaaaatcaa
     1141 acacaagaat ggtctggttc tggcacgtta acaaaagatg gtaaagttcg tttgttctac
     1201 acggattatt caggtaaaca atacggaaaa caaacgttaa cgacagctca agtgaacatg
     1261 tctcagccaa acgataatac gttaaaagtt gatggagtag aagattacaa gtcaattttt
     1321 gacggtgacg gaaagattta tcaaacggtt cagcagttta ttgatgaagg cggttatgac
     1381 acaggggata accatacgct aagagatcct cactacatag aagacaacgg acataagtac
     1441 cttgttttcg aagctaacac aggaacagaa gatggctatc aaggtgaaga ctctctatat
     1501 aacagagcat actacggagg aaacaatcct ttcttccaat ctgaaaagaa aaagcttctt
     1561 gaaggatcta ataaagaaaa agcttcttta gcaaacgggg ctttgggcat tattgagtta
     1621 aatgatgact atacgctcaa aaaagtaatg aagccactta ttacatcaaa cacagttaca
     1681 gacgaaattg aacgtgcaaa cattttcaaa aaagatggaa aatggtactt gttcaccgat
     1741 tctcgcggat ctaaaatgac gattgatgga atcggacaag atgatgtgta tatgctaggc
     1801 tatgtttcaa atacgttaac aggaaaatat aagcctttaa atgatacagg attagttctt
     1861 catatggatt tagatcctaa tgataaaaca tttacgtatt ctcactttgc tgtacctcaa
     1921 acaaaaggcg ataatgtcgt tattacaagc tacatgacaa atagaggctt ttatgaagac
     1981 aaccactcta catttgcacc aagcttctta gtgaacattg acggttcaaa aacatctgtt
     2041 gtaaaagatc gtgttcttga gcaaggtcag ttaacagtag acgaagactg agcgctttgc
     2101 aatagcggat ccggtaccgg ccgcatgccg gcggcacctc gctaacggat tcaccactcc
     2161 aagaattgga gccaatcaat tcttgcggag aactgtgaat gcgcaaacca acccttggca
     2221 gaacatatcc atcgcgtccg ccatctccag cagccgcacg cggcgcatct cgggccgcgt
     2281 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2341 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2401 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2461 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2521 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2581 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2641 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2701 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     2761 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2821 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2881 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2941 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3001 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3061 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3121 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3181 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3241 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3301 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3361 ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3421 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3481 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3541 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3601 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3661 cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3721 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3781 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     3841 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     3901 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     3961 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4021 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     4081 aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt cctgttataa aaaaaggatc
     4141 aattttgaac tctctcccaa agttgatccc ttaacgattt agaaatccct ttgagaatgt
     4201 ttatatacat tcaaggtaac cagccaacta atgacaatga ttcctgaaaa aagtaataac
     4261 aaattactat acagataagt tgactgatca acttccatag gtaacaacct ttgatcaagt
     4321 aagggtatgg ataataaacc acctacaatt gcaatacctg ttccctctga taaaaagctg
     4381 gtaaagttaa gcaaactcat tccagcacca gcttcctgct gtttcaagct acttgaaaca
     4441 attgttgata taactgtttt ggtgaacgaa agcccaccta aaacaaatac gattataatt
     4501 gtcatgaacc atgatgttgt ttctaaaaga aaggaagcag ttaaaaagct aacagaaaga
     4561 aatgtaactc cgatgtttaa cacgtataaa ggacctcttc tatcaacaag tatcccacca
     4621 atgtagccga aaataatgac actcattgtt ccagggaaaa taattacact tccgatttcg
     4681 gcagtactta gctggtgaac atctttcatc atataaggaa ccatagagac aaaccctgct
     4741 actgttccaa atataattcc cccacaaaga actccaatca taaaaggtat atttttccct
     4801 aatccgggat caacaaaagg atctgttact ttcctgatat gttttacaaa tatcaggaat
     4861 gacagcacgc taacgataag aaaagaaatg ctatatgatg ttgtaaacaa cataaaaaat
     4921 acaatgccta cagacattag tataattcct ttgatatcaa aatgaccttt tatccttact
     4981 tctttcttta ataatttcat aagaaacgga acagtgataa ttgttatcat aggaatgagt
     5041 agaagatagg accaatgaat ataatgggct atcattccac caatcgctgg accgactcct
     5101 tctcccatgg ctactatcga tccaataaga ccaaatgctt tacccctatt ttcctttgga
     5161 atatagcgcg caactacaac cattacgagt gctggaaatg cagctgcacc agccccttga
     5221 ataaaacgag ccataataag taaggaaaag aaagaatggc caacaaaccc aattaccgac
     5281 ccgaaacaat ttattataat tccaaatagg agtaaccttt tgatgcctaa ttgatcagat
     5341 agctttccat atacagctgt tccaatggaa aaggttaaca taaaggctgt gttcacccag
     5401 tttgtactcg caggtggttt attaaaatca tttgcaatat caggtaatga gacgttcaaa
     5461 accatttcat ttaatacgct aaaaaaagat aaaatgcaaa gccaaattaa aatttggttg
     5521 tgtcgtaaat tcgattgtga ataggatgta ttcacatttc accctccaat aatgagggca
     5581 gacgtagttt atagggttaa tgatacgctt ccctctttta attgaaccct gttacattca
     5641 ttacacttca taattaattc ctcctaaact tgattaaaac attttaccac atataaacta
     5701 agttttaaat tcagtatttc atcacttata caacaatatg gcccgtttgt tgaactactc
     5761 tttaataaaa taatttttcc gttcccaatt ccacattgca ataatagaaa atccatcttc
     5821 atcggctttt tcgtcatcat ctgtatgaat caaatcgcct tcttctgtgt catcaaggtt
     5881 taatttttta tgtatttctt ttaacaaacc accataggag attaaccttt tacggtgtaa
     5941 accttcctcc aaatcagaca aacgtttcaa attcttttct tcatcatcgg tcataaaatc
     6001 cgtatccttt acaggatatt ttgcagtttc gtcaattgcc gattgtatat ccgatttata
     6061 tttatttttc ggtcgaatca tttgaacttt tacatttgga tcatagtcta atttcattgc
     6121 ctttttccaa aattgaatcc attgtttttg attcacgtag ttttctgtat tcttaaaata
     6181 agttggttcc acacatacca atacatgcat gtgctgatta taagaattat ctttattatt
     6241 tattgtcact tccgttgcac gcataaaacc aacaagattt ttattaattt ttttatattg
     6301 catcattcgg cgaaatcctt gagccatatc tgacaaactc ttatttaatt cttcgccatc
     6361 ataaacattt ttaactgtta atgtgagaaa caaccaacga actgttggct tttgtttaat
     6421 aacttcagca acaacctttt gtgactgaat gccatgtttc attgctctcc tccagttgca
     6481 cattggacaa agcctggatt tacaaaacca cactcgatac aactttcttt cgcctgtttc
     6541 acgattttgt ttatactcta atatttcagc acaatctttt actctttcag cctttttaaa
     6601 ttcaagaata tgcagaagtt caaagtaatc aacattagcg attttctttt ctctccatgg
     6661 tctcactttt ccactttttg tcttgtccac taaaaccctt gatttttcat ctgaataaat
     6721 gctactatta ggacacataa tattaaaaga aacccccatc tatttagtta tttgtttggt
     6781 cacttataac tttaacagat ggggtttttc tgtgcaacca attttaaggg ttttcaatac
     6841 tttaaaacac atacatacca acacttcaac gcacctttca gcaactaaaa taaaaatgac
     6901 gttatttcta tatgtatcaa gataagaaag aacaagttca aaaccatcaa aaaaagacac
     6961 cttttcaggt gcttttttta ttttataaac tcattccctg atctcgactt cgttcttttt
     7021 ttacctctcg gttatgagtt agttcaaatt cgttcttttt aggttctaaa tcgtgttttt
     7081 cttggaattg tgctgtttta tcctttacct tgtctacaaa ccccttaaaa acgtttttaa
     7141 aggcttttaa gccgtctgta cgttcc
//
