LOCUS       sequence-69951-e        8094 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the gfp gene under control of the native xylose 
            inducible promoter of Bacillus megaterium.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8094)
  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 8094)
  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: pRBBm34
FEATURES             Location/Qualifiers
     source          1..8094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(211..213,214..927)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="ASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(241..269)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     864..884
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     rep_origin      complement(1109..1697)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1189..1208)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1868..2659,2660..2728))
                     /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     2491..2510
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2729..2833)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2901..2919
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(2997..4373)
                     /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(4573..5577)
                     /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"
     promoter        join(6002..6007,6008..6024,6025..6030)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 ttgaattaga tatttaaaag tatcatatct aatattataa ctaaattttc taaaaaaaac
       61 attgaaataa acatttattt tgtatatgat gagataaagt tagtttattg gataaacaaa
      121 ctaactcaat taagatagtt gatggataaa cttgttcact taaatcaaag ggggaaatgt
      181 acaatggtcc aaactagttc gaagatctgg gctagcaaag gagaagaact tttcactgga
      241 gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtcagt
      301 ggagagggtg aaggtgatgc tacatacgga aagcttaccc ttaaatttat ttgcactact
      361 ggaaaactac ctgttccatg gccaacactt gtcactactt tgacctatgg tgttcaatgc
      421 ttttcccgtt atccggatca tatgaaacgg catgactttt tcaagagtgc catgcccgaa
      481 ggttatgtac aggaacgcac tatatctttc aaagatgacg ggaactacaa gacgcgtgct
      541 gaagtcaagt ttgaaggtga tacccttgtt aatcgtatcg agttaaaagg tattgatttt
      601 aaagaagatg gaaacattct cggacacaaa ctcgagtaca actataactc acacaatgta
      661 tacatcacgg cagacaaaca aaagaatgga atcaaagcta acttcaaaat tcgccacaac
      721 attgaagatg gatccgttca actagcagac cattatcaac aaaatactcc aattggcgat
      781 ggccctgtcc ttttaccaga caaccattac ctgtcgacac aatctgccct ttcgaaagat
      841 cccaacgaaa agcgtgacca catggtcctt cttgagtttg taactgctgc tgggattaca
      901 catggcatgg atgagctcta caaataatga attcgcggcc gcatgccggc ggcacctcgc
      961 taacggattc accactccaa gaattggagc caatcaattc ttgcggagaa ctgtgaatgc
     1021 gcaaaccaac ccttggcaga acatatccat cgcgtccgcc atctccagca gccgcacgcg
     1081 gcgcatctcg ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1141 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1201 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1261 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1321 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1381 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1441 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1501 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     1561 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1621 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1681 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1741 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1801 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     1861 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     1921 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     1981 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     2041 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     2101 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     2161 tgcgcaacgt tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg
     2221 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     2281 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     2341 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     2401 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     2461 cgagttgctc ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa
     2521 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     2581 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     2641 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     2701 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     2761 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     2821 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     2881 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt caagaattcc
     2941 tgttataaaa aaaggatcaa ttttgaactc tctcccaaag ttgatccctt aacgatttag
     3001 aaatcccttt gagaatgttt atatacattc aaggtaacca gccaactaat gacaatgatt
     3061 cctgaaaaaa gtaataacaa attactatac agataagttg actgatcaac ttccataggt
     3121 aacaaccttt gatcaagtaa gggtatggat aataaaccac ctacaattgc aatacctgtt
     3181 ccctctgata aaaagctggt aaagttaagc aaactcattc cagcaccagc ttcctgctgt
     3241 ttcaagctac ttgaaacaat tgttgatata actgttttgg tgaacgaaag cccacctaaa
     3301 acaaatacga ttataattgt catgaaccat gatgttgttt ctaaaagaaa ggaagcagtt
     3361 aaaaagctaa cagaaagaaa tgtaactccg atgtttaaca cgtataaagg acctcttcta
     3421 tcaacaagta tcccaccaat gtagccgaaa ataatgacac tcattgttcc agggaaaata
     3481 attacacttc cgatttcggc agtacttagc tggtgaacat ctttcatcat ataaggaacc
     3541 atagagacaa accctgctac tgttccaaat ataattcccc cacaaagaac tccaatcata
     3601 aaaggtatat ttttccctaa tccgggatca acaaaaggat ctgttacttt cctgatatgt
     3661 tttacaaata tcaggaatga cagcacgcta acgataagaa aagaaatgct atatgatgtt
     3721 gtaaacaaca taaaaaatac aatgcctaca gacattagta taattccttt gatatcaaaa
     3781 tgacctttta tccttacttc tttctttaat aatttcataa gaaacggaac agtgataatt
     3841 gttatcatag gaatgagtag aagataggac caatgaatat aatgggctat cattccacca
     3901 atcgctggac cgactccttc tcccatggct actatcgatc caataagacc aaatgcttta
     3961 cccctatttt cctttggaat atagcgcgca actacaacca ttacgagtgc tggaaatgca
     4021 gctgcaccag ccccttgaat aaaacgagcc ataataagta aggaaaagaa agaatggcca
     4081 acaaacccaa ttaccgaccc gaaacaattt attataattc caaataggag taaccttttg
     4141 atgcctaatt gatcagatag ctttccatat acagctgttc caatggaaaa ggttaacata
     4201 aaggctgtgt tcacccagtt tgtactcgca ggtggtttat taaaatcatt tgcaatatca
     4261 ggtaatgaga cgttcaaaac catttcattt aatacgctaa aaaaagataa aatgcaaagc
     4321 caaattaaaa tttggttgtg tcgtaaattc gattgtgaat aggatgtatt cacatttcac
     4381 cctccaataa tgagggcaga cgtagtttat agggttaatg atacgcttcc ctcttttaat
     4441 tgaaccctgt tacattcatt acacttcata attaattcct cctaaacttg attaaaacat
     4501 tttaccacat ataaactaag ttttaaattc agtatttcat cacttataca acaatatggc
     4561 ccgtttgttg aactactctt taataaaata atttttccgt tcccaattcc acattgcaat
     4621 aatagaaaat ccatcttcat cggctttttc gtcatcatct gtatgaatca aatcgccttc
     4681 ttctgtgtca tcaaggttta attttttatg tatttctttt aacaaaccac cataggagat
     4741 taacctttta cggtgtaaac cttcctccaa atcagacaaa cgtttcaaat tcttttcttc
     4801 atcatcggtc ataaaatccg tatcctttac aggatatttt gcagtttcgt caattgccga
     4861 ttgtatatcc gatttatatt tatttttcgg tcgaatcatt tgaactttta catttggatc
     4921 atagtctaat ttcattgcct ttttccaaaa ttgaatccat tgtttttgat tcacgtagtt
     4981 ttctgtattc ttaaaataag ttggttccac acataccaat acatgcatgt gctgattata
     5041 agaattatct ttattattta ttgtcacttc cgttgcacgc ataaaaccaa caagattttt
     5101 attaattttt ttatattgca tcattcggcg aaatccttga gccatatctg acaaactctt
     5161 atttaattct tcgccatcat aaacattttt aactgttaat gtgagaaaca accaacgaac
     5221 tgttggcttt tgtttaataa cttcagcaac aaccttttgt gactgaatgc catgtttcat
     5281 tgctctcctc cagttgcaca ttggacaaag cctggattta caaaaccaca ctcgatacaa
     5341 ctttctttcg cctgtttcac gattttgttt atactctaat atttcagcac aatcttttac
     5401 tctttcagcc tttttaaatt caagaatatg cagaagttca aagtaatcaa cattagcgat
     5461 tttcttttct ctccatggtc tcacttttcc actttttgtc ttgtccacta aaacccttga
     5521 tttttcatct gaataaatgc tactattagg acacataata ttaaaagaaa cccccatcta
     5581 tttagttatt tgtttggtca cttataactt taacagatgg ggtttttctg tgcaaccaat
     5641 tttaagggtt ttcaatactt taaaacacat acataccaac acttcaacgc acctttcagc
     5701 aactaaaata aaaatgacgt tatttctata tgtatcaaga taagaaagaa caagttcaaa
     5761 accatcaaaa aaagacacct tttcaggtgc tttttttatt ttataaactc attccctgat
     5821 ctcgacttcg ttcttttttt acctctcggt tatgagttag ttcaaattcg ttctttttag
     5881 gttctaaatc gtgtttttct tggaattgtg ctgttttatc ctttaccttg tctacaaacc
     5941 ccttaaaaac gtttttaaag gcttttaagc cgtctgtacg ttccttaagg aattctcatg
     6001 tttgacagct tatcatcgat aagctttaat gcggtagttt atcacagtta aattgctaac
     6061 gcagtcaggc accgtgtatg aaatctaaca atgcgctcat cgtcatcctc ggcaccgtca
     6121 ccctggatgc tgtaggcata ggcttggtta tgccggtact gccgggcctc ttgcgggata
     6181 tcgtccattc cgacagcatc gccagtcact atggcgtgct gctagcgcta tatgcgttga
     6241 tgcaatttct atgcgcaccc gttctcggag cactgtccga ccgctttggc cgccgcccag
     6301 tcctgctcgc ttcgctactt ggagccacta tcgactacgc gatcatggcg accacacccg
     6361 tcctgtggat ctgacgcgtg taactgcgaa ggtaaaacgg gtgattgaaa ataaactaac
     6421 aaatgaagag gaacgaaaga gaagtctaga atttgttacg tttatatcgg atgaattact
     6481 gcaaaatgat acggagcctc gcacatttat tattcagaac tctcttttat cgctagagaa
     6541 aatccatact ctcgcacaga ccaaagaggt agaggaatat aaagaagtca taactactct
     6601 gacaagacat gattcataat ttgaaaagca ggtaaactaa cccaaaaggc aagactctcc
     6661 gtagtttaga aaagagcagg ttgctaataa catataaaca gccagttgcc gttatgatag
     6721 gtgactggct gcatgggatg aaaaggtgag ggtggagaca gacataacac tcttaataga
     6781 agagggtaat tctttctctt ttatagaaaa tcaattaatt gaaagtagct ccttcattct
     6841 taagatcaac gtgatatagg tttgctaacc tttgcgttca cttaactaac ttataggggt
     6901 aacacttaaa aaagaatcaa taacgataga aaccgctcct aaagcaggtg cattttttcc
     6961 taacgaagaa ggcaatagtt cacatttatt gtctaaatga gaatggactc tagaagaaac
     7021 ttcgttttta atcgtattta aaacaatggg atgagattca attatatgat ttctcaagat
     7081 aacagcttct atatcaaatg tattaaggat attggttaat ccaattccga tataaaagcc
     7141 aaagttttga agtgcattta acatttctac atcattttta tttgcgcgtt ccacaatctc
     7201 ttttcgagaa atattctttt cttctttaga gagcgaagcc agtaacgctt tttcagaagc
     7261 atataattcc caacagcctc gatttccaca gctgcatttg ggtccattaa aatctatcgt
     7321 catatgaccc atttccccag aaaaaccctg aacaccttta tacaattcgt tgttaataac
     7381 aagtccagtt ccaattccga tattaatact gatgtaaacg atgttttcat agttttttgt
     7441 cataccaaat actttttcac cgtatgctcc tgcattagct tcattttcaa caaaaaccgg
     7501 aacattaaac tcactctcaa ttaaaaactg caaatctttg atattccaat ttaagttagg
     7561 catgaaaata atttgctgat gacgatctac aaggcctgga acacaaattc ctattccgac
     7621 tagaccataa ggggactcag gcatatgggt tacaaaacca tgaataagtg caaataaaat
     7681 ctcttttact tcactagcgg aagaactaga caagtcagaa gtcttctcga gaataatatt
     7741 tccttctaag tcggttagaa ttccgttaag atagtcgact cctatatcaa taccaatcga
     7801 gtagcctgca ttcttattaa aaacaagcat tacaggtctt ctgccgcctc tagattgccc
     7861 tgccccaatt tcaaaaataa aatctttttc aagcagtgta tttacttgag aggagacagt
     7921 agacttgttt aatcctgtaa tctcagagag agttgccctg gagacagggg agttcttcaa
     7981 aatttcatct aatattaatt tttgattcat tttttttact aaagcttgat ctgcaatttg
     8041 aataataacc actcctttgt ttatccaccg aactaagttg gtgttttttg aagc
//
