LOCUS       sequence-226389-        5811 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 5811)
  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 5811)
  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-Tn5-131
FEATURES             Location/Qualifiers
     source          1..5811
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(420..422,423..1136)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(462..483)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(723..742)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1070..1091
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1150..1244
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1288..1947
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1354..1373)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2012..2098
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3842..3860)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3928..4032
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4033..4101,4102..4893)
                     /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(4251..4270)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            5132..5241
                     /note="incP origin of transfer"
     primer_bind     5795..5811
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5795..5811
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 ccccgggctg caggaattcg atatccataa actgccagga attggggatc ggattcgagc
       61 tcgcacgctg ccgcaagcac tcagggcgca agggctgcta aaggaagcgg aacacgtaga
      121 aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga
      181 caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat
      241 agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct
      301 ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg ccaaggatct
      361 gatggcgcag gggatcaaga tctgatcaag agacaggata ctagtggagg aagaaaaaaa
      421 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
      481 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
      541 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
      601 tgaccaccct gtcctggggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc
      661 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
      721 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
      781 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
      841 tggagtacaa ctactttagc gacaacgtct atatcaccgc cgacaagcag aagaacggca
      901 tcaaggccaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc
      961 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1021 tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1081 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaattag
     1141 ctgagcttgg actcctgttg atagatccag taatgacctc agaactccat ctggatttgt
     1201 tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaagc tagcttggcg
     1261 agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata taccaccgtt
     1321 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt
     1381 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat
     1441 aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa tgctcatccg
     1501 gaatttcgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt
     1561 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac
     1621 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg
     1681 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg
     1741 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc
     1801 accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt
     1861 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac
     1921 tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca gttattggtg cccttaaacg
     1981 cctggggtaa tgactctcta gcttgaggca tcaaataaaa cgaaaggctc agtcgaaaga
     2041 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     2101 gccgctagga gcttgcggcc gccaggatat cgacccaagt accgccacct aacaattcgt
     2161 tcaagccgag atcggcttcc cggccgacgc gtcaattcga gcgcgtcaat tcgagggccg
     2221 cggcctaggc ggccagatct gatcaagaga cagtctagct agaattaatt cgagtcgacc
     2281 gatcccgtac acaagtagcg tcctgaacgg aacctttccc gttttccagg atctgatctt
     2341 ccatgtgacc tcctaacatg gtaacgttca tgataacttc tgctcttcat cgtgcggccg
     2401 actgggctaa atctgtgttc tcttcggcgg cgctgggtga tcctcgccgt actgcccgct
     2461 tggttaacgt cgccgcccaa ttggcaaaat attctggtaa atcaataacc atctcatcag
     2521 agggtagtga agccatgcag gaaggcgctt accgatttat ccgcaatccc aacgtttctg
     2581 ccgaggcgat cagaaaggct ggcgccatgc aaacagtcaa gttggctcag gagtttcccg
     2641 aactgctggc cattgaggac accacctctt tgagttatcg ccaccaggtc gccgaagagc
     2701 ttggcaagct gggctctatt caggataaat cccgcggatg gtgggttcac tccgttctct
     2761 tgctcgaggc caccacattc cgcaccgtag gattactgca tcaggagtgg tggatgcgcc
     2821 cggatgaccc tgccgatgcg gatgaaaagg agagtggcaa atggctggca gccgccgcaa
     2881 ctagccggtt acgcatgggc agcatgatga gcaacgtgat tgcggtctgt gaccgcgaag
     2941 ccgatattca tgcttatctg caggacaaac tggcgcataa cgagcgcttc gtggtgcgct
     3001 ccaagcaccc acgcaaggac gtagagtctg ggttgtatct gtacgaccat ctgaagaacc
     3061 aaccggagtt gggtggctat cagatcagca ttccgcaaaa gggcgtggtg gataaacgcg
     3121 gtaaacgtaa aaatcgacca gcccgcaagg cgagcttgag cctgcgcagt gggcgcatca
     3181 cgctaaaaca ggggaatatc acgctcaacg cggtgctggc cgaggagatt aacccgccca
     3241 agggtgagac cccgttgaaa tggttgttgc tgaccagcga accggtcgag tcgctagccc
     3301 aagccttgcg cgtcatcgac atttataccc atcgctggcg gatcgaggag ttccataagg
     3361 catggaaaac cggagcagga gccgagaggc aacgcatgga ggagccggat aatctggagc
     3421 ggatggtctc gatcctctcg tttgttgcgg tcaggctgtt acagctcaga gaaagcttca
     3481 cgctgccgca agcactcagg gcgcaagggc tgctaaagga agcggaacac gtagaaagcc
     3541 agtccgcaga aacggtgctg accccggatg aatgtcagct actgggctat ctggacaagg
     3601 gaaaacgcaa gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg gcgatagcta
     3661 gactgggcgg ttttatggac agcaagcgaa ccggaattgc cagctggggc gccctctggg
     3721 aaggttggga agccctgcaa agtaaactgg atggctttct tgccgccaag gatctgatgg
     3781 cgcaggggat caagatctga tcggtcgatc gaattaattc ccaattcttg aagacgaaag
     3841 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     3901 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3961 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4021 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4081 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4141 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4201 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4261 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4321 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     4381 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     4441 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     4501 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     4561 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     4621 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     4681 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     4741 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     4801 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     4861 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     4921 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccgggga
     4981 attggggatc aattccgttt tccgctgcat aaccctgctt cggggtcatt atagcgattt
     5041 tttcggtata tccatccttt ttcgcacgat atacaggatt ttgccaaagg gttcgtgtag
     5101 actttccttg gtgtatccaa cggcgtcagc cgggcaggat aggtgaagta ggcccacccg
     5161 cgagcgggtg ttccttcttc actgtccctt attcgcacct ggcggtgctc aacgggaatc
     5221 ctgctctgcg aggctggccg gctaccgccg gcgtaacaga tgagggcaag cggatggctg
     5281 atgaaaccaa gccaaccagg aagggcagcc cacctatcac ggaattgatc cggccacgat
     5341 gcgtccggcg tagaggatct gaagatcagc agttcaacct gttgatagta cgtactaagc
     5401 tctcatgttt cacgtactaa gctctcatgt ttaacgtact aagctctcat gtttaacgaa
     5461 ctaaaccctc atggctaacg tactaagctc tcatgtttca cgtactaagc tctcatgttt
     5521 gaacaataaa attaatataa atcagcaact taaatagcct ctaaggtttt aagttttata
     5581 agaaaaaaaa gaatatataa ggcttttaaa gcttttaagg tttaacggtt gtggacaaca
     5641 agccagggat gtaacgcact gagaagccct tagagcctct caaagcaatt ttgagtgaca
     5701 caggaacact taacggctga catgggaatt aattctgact cttatacaca agtgcggccc
     5761 tcgaattgac gcgctccacc gcggtggcgg ccgctctaga actagtggat c
//
