LOCUS       sequence-261973-        6054 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent protein sfGFP(Sp); under control of mannose inducible 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6054)
  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 6054)
  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-sfGFP(Sp)-Pman
FEATURES             Location/Qualifiers
     source          1..6054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(148..150,151..864)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1018..1037)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     misc_feature    3904..4044
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4059..4076)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4230..4818)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4310..4329)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4989..5780,5781..5849))
                     /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     5612..5631
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5850..5954)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gaattcgcag ggattattcc ttgctttttt tgttataggg aaaaatgcct ttattaccgg
       61 aacctatggt aaaaaaagcg attttaatga gctgatttcg gtatacagtt gagacatcta
      121 gatgattaac taataaggag gacaaacatg tcaaaaggag aagaactttt tacaggtgta
      181 gtacctatct tggttgaatt ggatggtgat gttaacggtc acaaattttc tgtacgtggt
      241 gaaggtgaag gtgatgcaac taacggtaaa ttgacactta aattcatttg tacaactgga
      301 aaacttcctg ttccttggcc tactcttgtt acaacattga catatggagt acaatgtttt
      361 tcacgttatc ctgatcatat gaaacgtcac gattttttta aatctgctat gccagaaggt
      421 tatgtacaag aacgtacaat ttcatttaaa gatgacggaa catataaaac acgtgctgaa
      481 gtaaaattcg aaggtgacac tcttgttaat cgtatcgaat tgaaaggaat cgatttcaaa
      541 gaagatggta acattttggg acacaaactt gaatacaact tcaactctca taatgtttat
      601 atcacagctg acaaacaaaa aaacggtatt aaagctaatt ttaaaattcg tcacaatgtt
      661 gaagatggat ctgttcaatt ggctgatcat tatcaacaaa atacaccaat cggagacgga
      721 ccagtattgc ttccagataa ccactacctt tctactcaat cagttctttc aaaagatcct
      781 aacgaaaaac gtgaccatat ggtacttctt gaatttgtta cagcagcagg tatcactcac
      841 ggtatggacg aactttataa ataataatga gcactagtca aggtcggcaa ttctgcagta
      901 ctaggaatcc gccgagcttc gacgagattt tcaggagcta aggaagctaa aatggagaaa
      961 aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag
     1021 gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagaacaa agaatacaag
     1081 aaaatattta caaaaaatca atttaacaat tccttaaaac atgcaggaat tgacgattta
     1141 aacaatatta gctttgaaca attcttatct cttttcaata gctataaatt atttaataag
     1201 taagttaagg gatgcataaa ctgcatccct taacttgttt ttcgtgtgcc tattttttgt
     1261 gaatcgctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     1321 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     1381 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     1441 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     1501 actgagagtg caccatacaa aacatatttc aacacaatac aaatgggtta gttaaaaaag
     1561 caggccttct aaaggtctgc tttttttatt tgattatgta atttttaatg ccaggatgcc
     1621 aataagccat aacctcaaat gcaccatttg caacctcgtc atcttcttcc tcaatcttga
     1681 ccagatcgcc gtcctccgca tcaccaagat tcagctcttt atgtatctcc ttcaaaatgc
     1741 caccgtatcc aattaacctt cgagctgcca acgcatcatc caagtaaagc accgtgttca
     1801 gattgtcttc agtcacctta ttaccgcgca caacatccgt atccttaacc ggatatttag
     1861 agatttcgag aacagctttt tgctccatca ttgcgttccg cacatcgttt tcaatctgtt
     1921 cagcgtcaat cttagcttta cctttcactc gacgaatatc gacaattgga gtgtaatcca
     1981 atttcatcgc ctttttccaa aggctcgtcc actccgcctg cttaatatag tttttcccaa
     2041 aataattttt ccttactggt atcaacacat gaaaatgagg atgatatgta tcttcttcat
     2101 gatttttggt aatctctaaa gctctgaaaa atccaagaac cgaagttttt acttttttgt
     2161 actggaacag tttcctaaag ccttccatca tcgcagaaat ttgtggcttc agccgttctc
     2221 cctttacatt tcgaatcgtc agcgtgagaa aaatccatcc gcagccgtac tgtctattgg
     2281 cttcctctac gatcaactta ttgtgataag caatttttaa cgacctgcgc cacgcacaca
     2341 tcggacataa cctcacttta caaaaatggg cttgatacag ttttaacttg cccgtctccg
     2401 ggtctctctt aaacgaaaga tactctgcac aactaattag tttttcagcc tttttgccat
     2461 agtaaggtgc cccaatctta ctctctaacg cttcgtaatg ctccgccatg aggttcgtcc
     2521 gtctcttttt ccccttccaa tcccgctttt tacctgttgc ggttttatct tcgaggatgc
     2581 tataatcatt ttcagatgaa taaatcaaca aaaaaactcc ttctgagcta gttctctagc
     2641 attctattat tttgattcga caccttaata atagcagaag gagtttttac ctgtcaaaga
     2701 accatcaaac ccttgataca caaggctttg acctaatttt gaaaaatgat gttgtttcta
     2761 tatagtatca agataagaaa gaaaaggatt tttcgctacg ctcaaatcct ttaaaaaaac
     2821 acaaaagacc acatttttta atgtggtctt ttattcttca actaaagcac ccattagttc
     2881 aacaaacgaa aattggataa agtgggatat ttttaaaata tatatttatg ttacagtaat
     2941 attgactttt aaaaaaggat tgattctaat gaagaaagca gacaagtaag cctcctaaat
     3001 tcactttaga taaaaattta ggaggcatat caaatgaact ttaataaaat tgatttagac
     3061 aattggaaga gaaaagagat atttaatcat tatttgaacc aacaaacgac ttttagtata
     3121 accacagaaa ttgatattag tgttttatac cgaaacataa aacaagaagg atataaattt
     3181 taccctgcat ttattttctt agtgacaagg gtgataaact caaatacagc ttttagaact
     3241 ggttacaata gcgacggaga gttaggttat tgggataagt tagagccact ttatacaatt
     3301 tttgatggtg tatctaaaac attctctggt atttggactc ctgtaaagaa tgacttcaaa
     3361 gagttttatg atttatacct ttctgatgta gagaaatata atggttcggg gaaattgttt
     3421 cccaaaacac ctatacctga aaatgctttt tctctttcta ttattccatg gacttcattt
     3481 actgggttta acttaaatat caataataat agtaattacc ttctacccat tattacagca
     3541 ggaaaattca ttaataaagg taattcaata tatttaccgc tatctttaca ggtacatcat
     3601 tctgtttgtg atggttatca tgcaggattg tttatgaact ctattcagga attgtcagat
     3661 aggcctaatg actggctttt ataatatgag ataatgccga ctgtactttt tacagtcggt
     3721 tttctaatgt cactaacctg ccccgttagt cgccattcgc cactgcctcg cgcgtttcgg
     3781 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     3841 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     3901 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg
     3961 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     4021 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4081 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4141 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4201 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4261 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4321 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4381 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4441 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4501 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4561 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4621 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     4681 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4741 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4801 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4861 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4921 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4981 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5041 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5101 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5161 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5221 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5281 ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg
     5341 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5401 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5461 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5521 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5581 ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta
     5641 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5701 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5761 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5821 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5881 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5941 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     6001 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct tcaa
//
