LOCUS       sequence-261727-        6048 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent protein sfGFP(SPS6) variant for optimized expression in 
            Bacillus mycoides; under control of mannose inducible promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6048)
  AUTHORS   Yi Y, Frenzel E, Spoelder J, Elzenga JTM, van Elsas JD, Kuipers OP
  TITLE     Optimized fluorescent proteins for the rhizosphere-associated 
            bacterium Bacillus mycoides with endophytic and biocontrol agent 
            potential.
  JOURNAL   Environ Microbiol Rep. 2018 Feb;10(1):57-74. doi: 
            10.1111/1758-2229.12607. Epub 2018 Jan 5.
  PUBMED    29195004
REFERENCE   2  (bases 1 to 6048)
  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: 
            "10.1111/1758-2229.12607"; journalName: "Environ Microbiol Rep"; 
            date: "2018-02"; volume: "10"; issue: "1"; pages: "57-74"
COMMENT     Sequence Label: pAD-SPS6-Pman
FEATURES             Location/Qualifiers
     source          1..6048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1012..1031)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     misc_feature    3898..4038
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4053..4070)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4224..4812)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4304..4323)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4983..5774,5775..5843))
                     /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     5606..5625
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5844..5948)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gaattcgcag ggattattcc ttgctttttt tgttataggg aaaaatgcct ttattaccgg
       61 aacctatggt aaaaaaagcg attttaatga gctgatttcg gtatacagtt gagacatcta
      121 gaactaataa ggaggacaaa catgtcaaaa ggagaagaac tttttacagg tgtagtacct
      181 atcttggttg aattggatgg tgatgttaac ggtcacaaat tttctgtacg tggtgaaggt
      241 gaaggtgatg caactgacgg taaattgaca cttaaattca tttgtacaac tggaaaactt
      301 cctgttcctt ggcctactct tgttacaaca ttgacatatg gagtacaatg tttttcacgt
      361 tatcctgatc atatgaaacg tcacgatttt tttaaatctg ctatgccaga aggttatgta
      421 caagaacgta caatttcatt taaagatgac ggaacatata aaacacgtgc tgaagtaaaa
      481 ttcgaaggtg acactcttgt taatcgtatc gaattgaaag gaatcgattt caaagaagat
      541 ggtaacattt tgggacacaa acttgaatac aacttcaact ctcataatgt ttatatcaca
      601 gctgacaaac aaaaaaacgg tattaaagct aattttaaaa ttcgtcacaa tgttgaagat
      661 ggatctgttc aattggccga tcattatcaa caaaatacac caatcggaga cggaccagta
      721 ttgcttccag ataaccacta cctttctact caatcagttc tttcaaaaga tcctaacgaa
      781 aaacgtgacc atatggtact tcttgaattt gttacagcag caggtatcac tcacggtatg
      841 gacgaacttt ataaataata atgagcacta gtcaagggca tgctaggacg agtaggacaa
      901 atccgccgag cttcgacgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
      961 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
     1021 cagtcagttg ctcaatgtac ctataaccag accgttcaga acaaagaata caagaaaata
     1081 tttacaaaaa atcaatttaa caattcctta aaacatgcag gaattgacga tttaaacaat
     1141 attagctttg aacaattctt atctcttttc aatagctata aattatttaa taagtaagtt
     1201 aagggatgca taaactgcat cccttaactt gtttttcgtg tgcctatttt ttgtgaatcg
     1261 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     1321 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     1381 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     1441 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     1501 agtgcaccat acaaaacata tttcaacaca atacaaatgg gttagttaaa aaagcaggcc
     1561 ttctaaaggt ctgctttttt tatttgatta tgtaattttt aatgccagga tgccaataag
     1621 ccataacctc aaatgcacca tttgcaacct cgtcatcttc ttcctcaatc ttgaccagat
     1681 cgccgtcctc cgcatcacca agattcagct ctttatgtat ctccttcaaa atgccaccgt
     1741 atccaattaa ccttcgagct gccaacgcat catccaagta aagcaccgtg ttcagattgt
     1801 cttcagtcac cttattaccg cgcacaacat ccgtatcctt aaccggatat ttagagattt
     1861 cgagaacagc tttttgctcc atcattgcgt tccgcacatc gttttcaatc tgttcagcgt
     1921 caatcttagc tttacctttc actcgacgaa tatcgacaat tggagtgtaa tccaatttca
     1981 tcgccttttt ccaaaggctc gtccactccg cctgcttaat atagtttttc ccaaaataat
     2041 ttttccttac tggtatcaac acatgaaaat gaggatgata tgtatcttct tcatgatttt
     2101 tggtaatctc taaagctctg aaaaatccaa gaaccgaagt ttttactttt ttgtactgga
     2161 acagtttcct aaagccttcc atcatcgcag aaatttgtgg cttcagccgt tctcccttta
     2221 catttcgaat cgtcagcgtg agaaaaatcc atccgcagcc gtactgtcta ttggcttcct
     2281 ctacgatcaa cttattgtga taagcaattt ttaacgacct gcgccacgca cacatcggac
     2341 ataacctcac tttacaaaaa tgggcttgat acagttttaa cttgcccgtc tccgggtctc
     2401 tcttaaacga aagatactct gcacaactaa ttagtttttc agcctttttg ccatagtaag
     2461 gtgccccaat cttactctct aacgcttcgt aatgctccgc catgaggttc gtccgtctct
     2521 ttttcccctt ccaatcccgc tttttacctg ttgcggtttt atcttcgagg atgctataat
     2581 cattttcaga tgaataaatc aacaaaaaaa ctccttctga gctagttctc tagcattcta
     2641 ttattttgat tcgacacctt aataatagca gaaggagttt ttacctgtca aagaaccatc
     2701 aaacccttga tacacaaggc tttgacctaa ttttgaaaaa tgatgttgtt tctatatagt
     2761 atcaagataa gaaagaaaag gatttttcgc tacgctcaaa tcctttaaaa aaacacaaaa
     2821 gaccacattt tttaatgtgg tcttttattc ttcaactaaa gcacccatta gttcaacaaa
     2881 cgaaaattgg ataaagtggg atatttttaa aatatatatt tatgttacag taatattgac
     2941 ttttaaaaaa ggattgattc taatgaagaa agcagacaag taagcctcct aaattcactt
     3001 tagataaaaa tttaggaggc atatcaaatg aactttaata aaattgattt agacaattgg
     3061 aagagaaaag agatatttaa tcattatttg aaccaacaaa cgacttttag tataaccaca
     3121 gaaattgata ttagtgtttt ataccgaaac ataaaacaag aaggatataa attttaccct
     3181 gcatttattt tcttagtgac aagggtgata aactcaaata cagcttttag aactggttac
     3241 aatagcgacg gagagttagg ttattgggat aagttagagc cactttatac aatttttgat
     3301 ggtgtatcta aaacattctc tggtatttgg actcctgtaa agaatgactt caaagagttt
     3361 tatgatttat acctttctga tgtagagaaa tataatggtt cggggaaatt gtttcccaaa
     3421 acacctatac ctgaaaatgc tttttctctt tctattattc catggacttc atttactggg
     3481 tttaacttaa atatcaataa taatagtaat taccttctac ccattattac agcaggaaaa
     3541 ttcattaata aaggtaattc aatatattta ccgctatctt tacaggtaca tcattctgtt
     3601 tgtgatggtt atcatgcagg attgtttatg aactctattc aggaattgtc agataggcct
     3661 aatgactggc ttttataata tgagataatg ccgactgtac tttttacagt cggttttcta
     3721 atgtcactaa cctgccccgt tagtcgccat tcgccactgc ctcgcgcgtt tcggtgatga
     3781 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     3841 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
     3901 agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca
     3961 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     4021 agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4081 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4141 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4201 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4261 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4321 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4381 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4441 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4501 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4561 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4621 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     4681 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4741 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4801 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4861 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4921 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4981 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5041 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5101 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5161 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5221 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5281 aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5341 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5401 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5461 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5521 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5581 tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg
     5641 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5701 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5761 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5821 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5881 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5941 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     6001 acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaa
//
