LOCUS       sequence-227203-        6670 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  miniTn5 plasmid to deliver constitutively expressed fluorescent 
            protein genes in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6670)
  AUTHORS   Schlechter RO, Jun H, Bernach M, Oso S, Boyd E, Munoz-Lintz DA, 
            Dobson RCJ, Remus DM, Remus-Emsermann MNP
  TITLE     Chromatic Bacteria - A Broad Host-Range Plasmid and Chromosomal 
            Insertion Toolbox for Fluorescent Protein Expression in Bacteria.
  JOURNAL   Front Microbiol. 2018 Dec 12;9:3052. doi: 10.3389/fmicb.2018.03052. 
            eCollection 2018.
  PUBMED    30631309
REFERENCE   2  (bases 1 to 6670)
  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: "Front 
            Microbiol."; date: "2018-12-12"; volume: "9:3052. doi"; pages: " 
            10.3389/fmicb.2018.03052. eCollection 2018"
COMMENT     Sequence Label: pMRE-Tn5-157
FEATURES             Location/Qualifiers
     source          1..6670
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     547..568
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      630..724
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             768..1427
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(834..853)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1492..1578
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3322..3340)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3408..3512
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3513..3581,3582..4373)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3731..3750)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            4612..4721
                     /note="incP origin of transfer"
     primer_bind     5275..5291
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5275..5291
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             5408..6202
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     GQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDVGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             join(6564..6566,6567..6569,6570..6670,1..613)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(6609..6630)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 cgatgccacc aacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
       61 gccctggccc accctcgtga ccaccttcgg ctacggcgtg gcctgcttca gccgctaccc
      121 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
      181 gcgcaccatc tctttcaagg acgacggtac ctacaagacc cgcgccgagg tgaagttcga
      241 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
      301 catcctgggg cacaagctgg agtacaactt caacagccac tacgtctata tcacggccga
      361 caagcagaag aactgcatca aggctaactt caagatccgc cacaacgttg aggacggcag
      421 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
      481 gcccgacaac cactacctga gccatcagtc caagctgagc aaagacccca acgagaagcg
      541 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg attacacatg gcatggacga
      601 gctgtacaag taataattag ctgagcttgg actcctgttg atagatccag taatgacctc
      661 agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttattggtga
      721 gaatccaagc tagcttggcg agattttcag gagctaagga agctaaaatg gagaaaaaaa
      781 tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat
      841 ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt
      901 taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc
      961 gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa agacggtgag ctggtgatat
     1021 gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc
     1081 tctggagtga ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg
     1141 cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg
     1201 tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca
     1261 acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga
     1321 tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc
     1381 ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca
     1441 gttattggtg cccttaaacg cctggggtaa tgactctcta gcttgaggca tcaaataaaa
     1501 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     1561 ctcctgagta ggacaaatcc gccgctagga gcttgcggcc gccaggatat cgacccaagt
     1621 accgccacct aacaattcgt tcaagccgag atcggcttcc cggccgacgc gtcaattcga
     1681 gcgcgtcaat tcgagggccg cggcctaggc ggccagatct gatcaagaga cagtctagct
     1741 agaattaatt cgagtcgacc gatcccgtac acaagtagcg tcctgaacgg aacctttccc
     1801 gttttccagg atctgatctt ccatgtgacc tcctaacatg gtaacgttca tgataacttc
     1861 tgctcttcat cgtgcggccg actgggctaa atctgtgttc tcttcggcgg cgctgggtga
     1921 tcctcgccgt actgcccgct tggttaacgt cgccgcccaa ttggcaaaat attctggtaa
     1981 atcaataacc atctcatcag agggtagtga agccatgcag gaaggcgctt accgatttat
     2041 ccgcaatccc aacgtttctg ccgaggcgat cagaaaggct ggcgccatgc aaacagtcaa
     2101 gttggctcag gagtttcccg aactgctggc cattgaggac accacctctt tgagttatcg
     2161 ccaccaggtc gccgaagagc ttggcaagct gggctctatt caggataaat cccgcggatg
     2221 gtgggttcac tccgttctct tgctcgaggc caccacattc cgcaccgtag gattactgca
     2281 tcaggagtgg tggatgcgcc cggatgaccc tgccgatgcg gatgaaaagg agagtggcaa
     2341 atggctggca gccgccgcaa ctagccggtt acgcatgggc agcatgatga gcaacgtgat
     2401 tgcggtctgt gaccgcgaag ccgatattca tgcttatctg caggacaaac tggcgcataa
     2461 cgagcgcttc gtggtgcgct ccaagcaccc acgcaaggac gtagagtctg ggttgtatct
     2521 gtacgaccat ctgaagaacc aaccggagtt gggtggctat cagatcagca ttccgcaaaa
     2581 gggcgtggtg gataaacgcg gtaaacgtaa aaatcgacca gcccgcaagg cgagcttgag
     2641 cctgcgcagt gggcgcatca cgctaaaaca ggggaatatc acgctcaacg cggtgctggc
     2701 cgaggagatt aacccgccca agggtgagac cccgttgaaa tggttgttgc tgaccagcga
     2761 accggtcgag tcgctagccc aagccttgcg cgtcatcgac atttataccc atcgctggcg
     2821 gatcgaggag ttccataagg catggaaaac cggagcagga gccgagaggc aacgcatgga
     2881 ggagccggat aatctggagc ggatggtctc gatcctctcg tttgttgcgg tcaggctgtt
     2941 acagctcaga gaaagcttca cgctgccgca agcactcagg gcgcaagggc tgctaaagga
     3001 agcggaacac gtagaaagcc agtccgcaga aacggtgctg accccggatg aatgtcagct
     3061 actgggctat ctggacaagg gaaaacgcaa gcgcaaagag aaagcaggta gcttgcagtg
     3121 ggcttacatg gcgatagcta gactgggcgg ttttatggac agcaagcgaa ccggaattgc
     3181 cagctggggc gccctctggg aaggttggga agccctgcaa agtaaactgg atggctttct
     3241 tgccgccaag gatctgatgg cgcaggggat caagatctga tcggtcgatc gaattaattc
     3301 ccaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     3361 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     3421 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     3481 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3541 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3601 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3661 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     3721 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     3781 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3841 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3901 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     3961 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     4021 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     4081 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     4141 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     4201 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     4261 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     4321 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     4381 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     4441 tctaggtgaa gatccgggga attggggatc aattccgttt tccgctgcat aaccctgctt
     4501 cggggtcatt atagcgattt tttcggtata tccatccttt ttcgcacgat atacaggatt
     4561 ttgccaaagg gttcgtgtag actttccttg gtgtatccaa cggcgtcagc cgggcaggat
     4621 aggtgaagta ggcccacccg cgagcgggtg ttccttcttc actgtccctt attcgcacct
     4681 ggcggtgctc aacgggaatc ctgctctgcg aggctggccg gctaccgccg gcgtaacaga
     4741 tgagggcaag cggatggctg atgaaaccaa gccaaccagg aagggcagcc cacctatcac
     4801 ggaattgatc cggccacgat gcgtccggcg tagaggatct gaagatcagc agttcaacct
     4861 gttgatagta cgtactaagc tctcatgttt cacgtactaa gctctcatgt ttaacgtact
     4921 aagctctcat gtttaacgaa ctaaaccctc atggctaacg tactaagctc tcatgtttca
     4981 cgtactaagc tctcatgttt gaacaataaa attaatataa atcagcaact taaatagcct
     5041 ctaaggtttt aagttttata agaaaaaaaa gaatatataa ggcttttaaa gcttttaagg
     5101 tttaacggtt gtggacaaca agccagggat gtaacgcact gagaagccct tagagcctct
     5161 caaagcaatt ttgagtgaca caggaacact taacggctga catgggaatt aattctgact
     5221 cttatacaca agtgcggccc tcgaattgac gcgctccacc gcggtggcgg ccgctctaga
     5281 actagtggat cccccgggct gcaggaattc gatatccata aactgccagg aattggggat
     5341 cggattcgag ctaagaattc ccttggggta tctttaaata ctgtagaaaa gaggaaggaa
     5401 ataataaatg gctaaaatga gaatatcacc ggaattgaaa aaactgatcg aaaaataccg
     5461 ctgcgtaaaa gatacggaag gaatgtctcc tgctaaggta tataagctgg tgggagaaaa
     5521 tgaaaaccta tatttaaaaa tgacggacag ccggtataaa gggaccacct atgatgtgga
     5581 acgggaaaag gacatgatgc tatggctgga aggaaagctg cctgttccaa aggtcctgca
     5641 ctttgaacgg catgatggct ggagcaatct gctcatgagt gaggccgatg gcgtcctttg
     5701 ctcggaagag tatgaagatg gacaaagccc tgaaaagatt atcgagctgt atgcggagtg
     5761 catcaggctc tttcactcca tcgacatatc ggattgtccc tatacgaata gcttagacag
     5821 ccgcttagcc gaattggatt acttactgaa taacgatctg gccgatgtgg attgcgaaaa
     5881 ctgggaagaa gacactccat ttaaagatcc gcgcgagctg tatgattttt taaagacgga
     5941 aaagcccgaa gaggaacttg tcttttccca cggcgacctg ggagacagca acatctttgt
     6001 gaaagatggc aaagtaagtg gctttattga tcttgggaga agcggcaggg cggacaagtg
     6061 gtatgacatt gccttctgcg tccggtcgat cagggaggat gtcggggaag aacagtatgt
     6121 cgagctattt tttgacttac tggggatcaa gcctgattgg gagaaaataa aatattatat
     6181 tttactggat gaattgtttt agagctcgca cgctgccgca agcactcagg gcgcaagggc
     6241 tgctaaagga agcggaacac gtagaaagcc agtccgcaga aacggtgctg accccggatg
     6301 aatgtcagct actgggctat ctggacaagg gaaaacgcaa gcgcaaagag aaagcaggta
     6361 gcttgcagtg ggcttacatg gcgatagcta gactgggcgg ttttatggac agcaagcgaa
     6421 ccggaattgc cagctggggc gccctctggt aaggttggga agccctgcaa agtaaactgg
     6481 atggctttct tgccgccaag gatctgatgg cgcaggggat caagatctga tcaagagaca
     6541 ggatactagt ggaggaagaa aaaatggtga gcaagggcga ggagctgttc accggggtgg
     6601 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtccgcggcg
     6661 agggcgaggg
//
