LOCUS       sequence-227216-        6650 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 6650)
  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 6650)
  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-Tn5-155
FEATURES             Location/Qualifiers
     source          1..6650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      258..352
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             396..1055
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(462..481)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1120..1206
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2950..2968)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3036..3140
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3141..3209,3210..4001)
                     /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(3359..3378)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            4240..4349
                     /note="incP origin of transfer"
     primer_bind     4903..4919
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4903..4919
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             5036..5830
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     GQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDVGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             join(6192..6650,1..237)
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(6325..6343)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     6593..6612
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 gaggacggcg tgctgaaggg cgacattaag atggccctgc gcctgaagga cggcggccgc
       61 tacctggcgg acttcaagac cacctacaag gccaagaagc ccgtgcagat gcccggcgcc
      121 tacaacgtcg accgcaagtt ggacatcacc tcccacaacg aggactacac cgtggtggaa
      181 cagtacgaac gctccgaggg ccgccactcc accggcggca tggacgagct gtacaagtaa
      241 gtaattagct gagcttggac tcctgttgat agatccagta atgacctcag aactccatct
      301 ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagcta
      361 gcttggcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata
      421 ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg
      481 ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa
      541 agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg
      601 ctcatccgga atttcgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc
      661 acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat
      721 accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg
      781 aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc
      841 cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc
      901 ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga
      961 ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac
     1021 aacagtactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt tattggtgcc
     1081 cttaaacgcc tggggtaatg actctctagc ttgaggcatc aaataaaacg aaaggctcag
     1141 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     1201 acaaatccgc cgctaggagc ttgcggccgc caggatatcg acccaagtac cgccacctaa
     1261 caattcgttc aagccgagat cggcttcccg gccgacgcgt caattcgagc gcgtcaattc
     1321 gagggccgcg gcctaggcgg ccagatctga tcaagagaca gtctagctag aattaattcg
     1381 agtcgaccga tcccgtacac aagtagcgtc ctgaacggaa cctttcccgt tttccaggat
     1441 ctgatcttcc atgtgacctc ctaacatggt aacgttcatg ataacttctg ctcttcatcg
     1501 tgcggccgac tgggctaaat ctgtgttctc ttcggcggcg ctgggtgatc ctcgccgtac
     1561 tgcccgcttg gttaacgtcg ccgcccaatt ggcaaaatat tctggtaaat caataaccat
     1621 ctcatcagag ggtagtgaag ccatgcagga aggcgcttac cgatttatcc gcaatcccaa
     1681 cgtttctgcc gaggcgatca gaaaggctgg cgccatgcaa acagtcaagt tggctcagga
     1741 gtttcccgaa ctgctggcca ttgaggacac cacctctttg agttatcgcc accaggtcgc
     1801 cgaagagctt ggcaagctgg gctctattca ggataaatcc cgcggatggt gggttcactc
     1861 cgttctcttg ctcgaggcca ccacattccg caccgtagga ttactgcatc aggagtggtg
     1921 gatgcgcccg gatgaccctg ccgatgcgga tgaaaaggag agtggcaaat ggctggcagc
     1981 cgccgcaact agccggttac gcatgggcag catgatgagc aacgtgattg cggtctgtga
     2041 ccgcgaagcc gatattcatg cttatctgca ggacaaactg gcgcataacg agcgcttcgt
     2101 ggtgcgctcc aagcacccac gcaaggacgt agagtctggg ttgtatctgt acgaccatct
     2161 gaagaaccaa ccggagttgg gtggctatca gatcagcatt ccgcaaaagg gcgtggtgga
     2221 taaacgcggt aaacgtaaaa atcgaccagc ccgcaaggcg agcttgagcc tgcgcagtgg
     2281 gcgcatcacg ctaaaacagg ggaatatcac gctcaacgcg gtgctggccg aggagattaa
     2341 cccgcccaag ggtgagaccc cgttgaaatg gttgttgctg accagcgaac cggtcgagtc
     2401 gctagcccaa gccttgcgcg tcatcgacat ttatacccat cgctggcgga tcgaggagtt
     2461 ccataaggca tggaaaaccg gagcaggagc cgagaggcaa cgcatggagg agccggataa
     2521 tctggagcgg atggtctcga tcctctcgtt tgttgcggtc aggctgttac agctcagaga
     2581 aagcttcacg ctgccgcaag cactcagggc gcaagggctg ctaaaggaag cggaacacgt
     2641 agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct
     2701 ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc
     2761 gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc
     2821 cctctgggaa ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga
     2881 tctgatggcg caggggatca agatctgatc ggtcgatcga attaattccc aattcttgaa
     2941 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     3001 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3061 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3121 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     3181 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3241 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3301 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     3361 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     3421 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     3481 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3541 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3601 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3661 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     3721 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     3781 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3841 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3901 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3961 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     4021 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4081 tccggggaat tggggatcaa ttccgttttc cgctgcataa ccctgcttcg gggtcattat
     4141 agcgattttt tcggtatatc catccttttt cgcacgatat acaggatttt gccaaagggt
     4201 tcgtgtagac tttccttggt gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg
     4261 cccacccgcg agcgggtgtt ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa
     4321 cgggaatcct gctctgcgag gctggccggc taccgccggc gtaacagatg agggcaagcg
     4381 gatggctgat gaaaccaagc caaccaggaa gggcagccca cctatcacgg aattgatccg
     4441 gccacgatgc gtccggcgta gaggatctga agatcagcag ttcaacctgt tgatagtacg
     4501 tactaagctc tcatgtttca cgtactaagc tctcatgttt aacgtactaa gctctcatgt
     4561 ttaacgaact aaaccctcat ggctaacgta ctaagctctc atgtttcacg tactaagctc
     4621 tcatgtttga acaataaaat taatataaat cagcaactta aatagcctct aaggttttaa
     4681 gttttataag aaaaaaaaga atatataagg cttttaaagc ttttaaggtt taacggttgt
     4741 ggacaacaag ccagggatgt aacgcactga gaagccctta gagcctctca aagcaatttt
     4801 gagtgacaca ggaacactta acggctgaca tgggaattaa ttctgactct tatacacaag
     4861 tgcggccctc gaattgacgc gctccaccgc ggtggcggcc gctctagaac tagtggatcc
     4921 cccgggctgc aggaattcga tatccataaa ctgccaggaa ttggggatcg gattcgagct
     4981 aagaattccc ttggggtatc tttaaatact gtagaaaaga ggaaggaaat aataaatggc
     5041 taaaatgaga atatcaccgg aattgaaaaa actgatcgaa aaataccgct gcgtaaaaga
     5101 tacggaagga atgtctcctg ctaaggtata taagctggtg ggagaaaatg aaaacctata
     5161 tttaaaaatg acggacagcc ggtataaagg gaccacctat gatgtggaac gggaaaagga
     5221 catgatgcta tggctggaag gaaagctgcc tgttccaaag gtcctgcact ttgaacggca
     5281 tgatggctgg agcaatctgc tcatgagtga ggccgatggc gtcctttgct cggaagagta
     5341 tgaagatgga caaagccctg aaaagattat cgagctgtat gcggagtgca tcaggctctt
     5401 tcactccatc gacatatcgg attgtcccta tacgaatagc ttagacagcc gcttagccga
     5461 attggattac ttactgaata acgatctggc cgatgtggat tgcgaaaact gggaagaaga
     5521 cactccattt aaagatccgc gcgagctgta tgatttttta aagacggaaa agcccgaaga
     5581 ggaacttgtc ttttcccacg gcgacctggg agacagcaac atctttgtga aagatggcaa
     5641 agtaagtggc tttattgatc ttgggagaag cggcagggcg gacaagtggt atgacattgc
     5701 cttctgcgtc cggtcgatca gggaggatgt cggggaagaa cagtatgtcg agctattttt
     5761 tgacttactg gggatcaagc ctgattggga gaaaataaaa tattatattt tactggatga
     5821 attgttttag agctcgcacg ctgccgcaag cactcagggc gcaagggctg ctaaaggaag
     5881 cggaacacgt agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac
     5941 tgggctatct ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg
     6001 cttacatggc gatagctaga ctgggcggtc ttatggacag caagcgaacc ggaattgcca
     6061 gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg
     6121 ccgccaagga tctgatggcg caggggatca agatctgatc aagagacagg atactagtgg
     6181 aggaagaaaa aatggtgagc aagggcgagg cagtgatcaa ggagttcatg cggttcaagg
     6241 tgcacatgga gggctccatg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     6301 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct
     6361 tctcctggga catcctgtcc cctcagttca tgtacggctc cagggccttc atcaagcacc
     6421 ccgccgacat ccccgactac tataagcagt ccttccccga gggcttcaag tgggagcgcg
     6481 tgatgaactt cgaggacggc ggcgccgtga ccgtgaccca ggacacctcc ctggaggacg
     6541 gcaccctgat ctacaaggtg aagctccgcg gcaccaactt ccctcctgac ggccccgtaa
     6601 tgcagaagaa gacaatgggc tgggaagcgt ccaccgagcg gttgtacccc
//
