LOCUS       sequence-257253-        7237 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent protein mKate(KPS12) variant for optimized expression in
            Bacillus mycoides..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7237)
  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 7237)
  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-mKate(KPS12)
FEATURES             Location/Qualifiers
     source          1..7237
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2201..2220)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     misc_feature    5087..5227
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5242..5259)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5413..6001)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5493..5512)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6172..6963,6964..7032))
                     /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     6795..6814
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7033..7137)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gaattcgagc tcggtacccg gggatcgcgt gaaagaaatt ttcggcgcag agcacgtaaa
       61 tgttcaacca cactctggtg cacaagcgaa catggcagta tacttcacga ttttagagca
      121 aggcgataca gtacttggta tgaatttatc tcatggtggt cacttaacac acggaagccc
      181 tgttaacttc agtggagtac aatataattt cgtagaatat ggcgtggatg ctgactctca
      241 ccgtattaat tacgatgatg tattagcaaa agcgaaagaa cataaaccaa aattaatcgt
      301 tgcaggtgca agtgcatacc ctcgtgttat cgatttcaag cgattccgtg agattgcaga
      361 tgaagtgggc gcttatttaa tggttgatat ggcacatatc gctggtttag tagctgctgg
      421 tttacatcca aatccagtac cacatgcaca tttcgttaca acgacaacac ataaaacgtt
      481 acgtggcccg cgtggtggta tgattttatg tgaagagcaa tttgcaaaac aaattgataa
      541 atcaatcttc cctggtattc aaggtggtcc acttatgcac gtaatcgctg caaaagctgt
      601 tgcgtttggt gaagcacttc aagatgattt caaaacatat gcacaaaata tcattaacaa
      661 tgcgaaccgc ttagctgaag gtcttcaaaa agaaggactt acacttgttt ctggcggaac
      721 agacaatcac ttaatcttga ttgatgttcg taacttagaa atcacaggta aagtagcaga
      781 gcacgtatta gatgaagttg gtattacagt gaacaaaaat acaattccat ttgaaacagc
      841 aagcccattt gtaacaagtg gtgtacgtat cggtacagca gctgtaacat ctcgtggttt
      901 cggtttagaa gaaatggatg aaattgcgtc acttattgct tatacattaa aaaatcatga
      961 aaatgaagct gcattagaag aagtacgtaa gcgtgtagaa gcgttaacta gcaaatttcc
     1021 aatgtatcca aatctataat agattgaaga agactgccga gacttaattg ttttggcggt
     1081 cttttttgtg gacatatatt atttttaaag tatgtataca aatgatgaat aaattttggc
     1141 gatataatga aggatacagc tcccataatt ggtaaagata ctagatagat tcatcgtaaa
     1201 atcatgattt tgccaaattt gcccttgaat attagtagcg ttttctttac aatcgtaaat
     1261 agtgtaaaaa agcgtgcaaa cgcatgaata tcatctaaag gagagattca catgggaaaa
     1321 ctgtatgtat ttgatcctct agaggaaaat taatgtcaga acttatcaag gaaaatatgc
     1381 acatgaaatt gtacatggaa ggaacagtaa ataatcacca ctttaaatgt acctcagaag
     1441 gagaaggaaa accatatgaa ggtactcaaa ccatgcgtat taaggccgtt gaaggtggac
     1501 cattgccttt tgcctttgat attcttgcta catcttttat gtacggatca aagactttta
     1561 tcaatcatac ccaaggtatc ccagatttct ttaaacagtc atttcctgaa ggatttacat
     1621 gggaacgtgt cacaacttat gaagatggtg gagtattgac agcaactcaa gatacatctt
     1681 tgcaagatgg ttgtcttatc tacaatgtaa agatccgtgg agttaatttt ccaagtaatg
     1741 gtcctgttat gcagaaaaag acccttggat gggaagcatc taccgaaaca ttatatcctg
     1801 ccgatggtgg attggaaggt cgtgctgata tggcattgaa acttgtcggt ggaggtcacc
     1861 ttatctgtaa tttgaagacc acataccgtt ctaaaaagcc agctaaaaat cttaagatgc
     1921 ctggtgttta ctacgtcgat cgtcgtttag aacgtatcaa agaagcatat aaggaaactt
     1981 atgttgaaca gcacgaagta gccgtcgcac ggtattgtga tttgcctagt aaattgggac
     2041 accgttaata ataaaagctt gagtaggcat gcaagcttga gtaggacaaa tccgccgagc
     2101 ttcgacgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac
     2161 caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc
     2221 tcaatgtacc tataaccaga ccgttcagaa caaagaatac aagaaaatat ttacaaaaaa
     2281 tcaatttaac aattccttaa aacatgcagg aattgacgat ttaaacaata ttagctttga
     2341 acaattctta tctcttttca atagctataa attatttaat aagtaagtta agggatgcat
     2401 aaactgcatc ccttaacttg tttttcgtgt gcctattttt tgtgaatcgc taagaaacca
     2461 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     2521 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     2581 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     2641 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     2701 caaaacatat ttcaacacaa tacaaatggg ttagttaaaa aagcaggcct tctaaaggtc
     2761 tgcttttttt atttgattat gtaattttta atgccaggat gccaataagc cataacctca
     2821 aatgcaccat ttgcaacctc gtcatcttct tcctcaatct tgaccagatc gccgtcctcc
     2881 gcatcaccaa gattcagctc tttatgtatc tccttcaaaa tgccaccgta tccaattaac
     2941 cttcgagctg ccaacgcatc atccaagtaa agcaccgtgt tcagattgtc ttcagtcacc
     3001 ttattaccgc gcacacaatc cgtatcctta accggatatt tagagatttc gcgaacagct
     3061 ttttgctcca tcattgcgtt ccgcacatcg ttttcaatct gttcagcgtc aatcttagct
     3121 ttacctttca ctcgacgaat atcgacaatt ggagtgtaat ccaatttcat cgcctttttc
     3181 caaaggctcg tccactccgc ctgcttaata tagtttttcc caaaataatt tttccttact
     3241 ggtatcaaca catgaaaatg aggatgatat gtatcttctt catgattttt ggtaatctct
     3301 aaagctctga aaaatccaag aaccgaagtt tttacttttt tgtactggaa cagtttccta
     3361 aagccttcca tcatcgcaga aatttgtggc ttcagccgtt ctccctttac atttcgaatc
     3421 gtcagcgtga gaaaaatcca tccgcagccg tactgtctat tggcttcctc tacgatcaac
     3481 ttattgtgat aagcaatttt taacgacctg cgccacgcac acatcggaca taacctcact
     3541 ttacaaaaat gggcttgata cagttttaac ttgcccgtct ccgggtctct cttaaacgaa
     3601 agatactctg cacaactaat tagtttttca gcctttttgc catagtaagg tgccccaatc
     3661 ttactctcta acgcttcgta atgctccgcc atgaggttcg tccgtctctt tttccccttc
     3721 caatcccgct ttttacctgt tgcggtttta tcttcgagga tgctataatc attttcagat
     3781 gaataaatca acaaaaaaac tccttctgag ctagttctct agcattctat tattttgatt
     3841 cgacacctta ataatagcag aaggagtttt tacctgtcaa agaaccatca aacccttgat
     3901 acacaaggct ttgacctaat tttgaaaaat gatgttgttt ctatatagta tcaagataag
     3961 aaagaaaagg atttttcgct acgctcaaat cctttaaaaa aacacaaaag accacatttt
     4021 ttaatgtggt ctttattctt caactaaagc acccattagt tcaacaaacg aaaattggat
     4081 aaagtgggat atttttaaaa tatatattta tgttacagta atattgactt ttaaaaaagg
     4141 attgattcta atgaagaaag cagacaagta agcctcctaa attcacttta gataaaaatt
     4201 taggaggcat atcaaatgaa ctttaataaa attgatttag acaattggaa gagaaaagag
     4261 atatttaatc attatttgaa ccaacaaacg acttttagta taaccacaga aattgatatt
     4321 agtgttttat accgaaacat aaaacaagaa ggatataaat tttaccctgc atttattttc
     4381 ttagtgacaa gggtgataaa ctcaaataca gcttttagaa ctggttacaa tagcgacgga
     4441 gagttaggtt attgggataa gttagagcca ctttatacaa tttttgatgg tgtatctaaa
     4501 acattctctg gtatttggac tcctgtaaag aatgacttca aagagtttta tgatttatac
     4561 ctttctgatg tagagaaata taatggttcg gggaaattgt ttcccaaaac acctatacct
     4621 gaaaatgctt tttctctttc tattattcca tggacttcat ttactgggtt taacttaaat
     4681 atcaataata atagtaatta ccttctaccc attattacag caggaaaatt cattaataaa
     4741 ggtaattcaa tatatttacc gctatcttta caggtacatc attctgtttg tgatggttat
     4801 catgcaggat tgtttatgaa ctctattcag gaattgtcag ataggcctaa tgactggctt
     4861 ttataatatg agataatgcc gactgtactt tttacagtcg gttttctaat gtcactaacc
     4921 tgccccgtta gtcgccattc gccagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc
     4981 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     5041 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc
     5101 agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt
     5161 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     5221 catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5281 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5341 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5401 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5461 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5521 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5581 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     5641 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5701 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5761 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5821 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     5881 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5941 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6001 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6061 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6121 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6181 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6241 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6301 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6361 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6421 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6481 cattgctgca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6541 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6601 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6661 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6721 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6781 ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6841 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6901 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6961 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7021 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7081 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7141 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     7201 taaaaatagg cgtatcacga ggccctttcg tcttcaa
//
