LOCUS       sequence-227076-        6661 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 6661)
  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 6661)
  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-Tn5-154
FEATURES             Location/Qualifiers
     source          1..6661
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..711
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     terminator      728..822
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             866..1525
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(932..951)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1590..1676
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3420..3438)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3506..3610
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3611..3679,3680..4471)
                     /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(3829..3848)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            4710..4819
                     /note="incP origin of transfer"
     primer_bind     5373..5389
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5373..5389
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             5506..6300
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     GQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDVGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 atggtgagca agggcgagga gaataacatg gccatcatca aggagttcat gcgcttcaag
       61 gtgcgcatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
      121 cgcccctacg agggctttca gaccgctaag ctgaaggtga ccaagggtgg ccccctgccc
      181 ttcgcctggg acatcctgtc ccctcatttc acctacggct ccaaggccta cgtgaagcac
      241 cccgccgaca tccccgacta cttcaagctg tccttccccg agggcttcaa gtgggagcgc
      301 gtgatgaact acgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
      361 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgtg
      421 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggt
      481 gccctgaagg gcaagatcaa gatgaggctg aagctgaagg acggcggcca ctacacctcc
      541 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacatcgtc
      601 gacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
      661 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta ataattagct
      721 gagcttggac tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc
      781 agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagcta gcttggcgag
      841 attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga
      901 tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac
      961 ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa
     1021 gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga
     1081 atttcgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta
     1141 caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga
     1201 tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc
     1261 ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag
     1321 tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac
     1381 catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca
     1441 tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg
     1501 cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt tattggtgcc cttaaacgcc
     1561 tggggtaatg actctctagc ttgaggcatc aaataaaacg aaaggctcag tcgaaagact
     1621 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     1681 cgctaggagc ttgcggccgc caggatatcg acccaagtac cgccacctaa caattcgttc
     1741 aagccgagat cggcttcccg gccgacgcgt caattcgagc gcgtcaattc gagggccgcg
     1801 gcctaggcgg ccagatctga tcaagagaca gtctagctag aattaattcg agtcgaccga
     1861 tcccgtacac aagtagcgtc ctgaacggaa cctttcccgt tttccaggat ctgatcttcc
     1921 atgtgacctc ctaacatggt aacgttcatg ataacttctg ctcttcatcg tgcggccgac
     1981 tgggctaaat ctgtgttctc ttcggcggcg ctgggtgatc ctcgccgtac tgcccgcttg
     2041 gttaacgtcg ccgcccaatt ggcaaaatat tctggtaaat caataaccat ctcatcagag
     2101 ggtagtgaag ccatgcagga aggcgcttac cgatttatcc gcaatcccaa cgtttctgcc
     2161 gaggcgatca gaaaggctgg cgccatgcaa acagtcaagt tggctcagga gtttcccgaa
     2221 ctgctggcca ttgaggacac cacctctttg agttatcgcc accaggtcgc cgaagagctt
     2281 ggcaagctgg gctctattca ggataaatcc cgcggatggt gggttcactc cgttctcttg
     2341 ctcgaggcca ccacattccg caccgtagga ttactgcatc aggagtggtg gatgcgcccg
     2401 gatgaccctg ccgatgcgga tgaaaaggag agtggcaaat ggctggcagc cgccgcaact
     2461 agccggttac gcatgggcag catgatgagc aacgtgattg cggtctgtga ccgcgaagcc
     2521 gatattcatg cttatctgca ggacaaactg gcgcataacg agcgcttcgt ggtgcgctcc
     2581 aagcacccac gcaaggacgt agagtctggg ttgtatctgt acgaccatct gaagaaccaa
     2641 ccggagttgg gtggctatca gatcagcatt ccgcaaaagg gcgtggtgga taaacgcggt
     2701 aaacgtaaaa atcgaccagc ccgcaaggcg agcttgagcc tgcgcagtgg gcgcatcacg
     2761 ctaaaacagg ggaatatcac gctcaacgcg gtgctggccg aggagattaa cccgcccaag
     2821 ggtgagaccc cgttgaaatg gttgttgctg accagcgaac cggtcgagtc gctagcccaa
     2881 gccttgcgcg tcatcgacat ttatacccat cgctggcgga tcgaggagtt ccataaggca
     2941 tggaaaaccg gagcaggagc cgagaggcaa cgcatggagg agccggataa tctggagcgg
     3001 atggtctcga tcctctcgtt tgttgcggtc aggctgttac agctcagaga aagcttcacg
     3061 ctgccgcaag cactcagggc gcaagggctg ctaaaggaag cggaacacgt agaaagccag
     3121 tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct ggacaaggga
     3181 aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc gatagctaga
     3241 ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc cctctgggaa
     3301 ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga tctgatggcg
     3361 caggggatca agatctgatc ggtcgatcga attaattccc aattcttgaa gacgaaaggg
     3421 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     3481 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3541 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3601 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3661 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3721 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3781 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3841 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3901 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3961 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4021 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4081 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4141 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4201 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     4261 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4321 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4381 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4441 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4501 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tccggggaat
     4561 tggggatcaa ttccgttttc cgctgcataa ccctgcttcg gggtcattat agcgattttt
     4621 tcggtatatc catccttttt cgcacgatat acaggatttt gccaaagggt tcgtgtagac
     4681 tttccttggt gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg cccacccgcg
     4741 agcgggtgtt ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa cgggaatcct
     4801 gctctgcgag gctggccggc taccgccggc gtaacagatg agggcaagcg gatggctgat
     4861 gaaaccaagc caaccaggaa gggcagccca cctatcacgg aattgatccg gccacgatgc
     4921 gtccggcgta gaggatctga agatcagcag ttcaacctgt tgatagtacg tactaagctc
     4981 tcatgtttca cgtactaagc tctcatgttt aacgtactaa gctctcatgt ttaacgaact
     5041 aaaccctcat ggctaacgta ctaagctctc atgtttcacg tactaagctc tcatgtttga
     5101 acaataaaat taatataaat cagcaactta aatagcctct aaggttttaa gttttataag
     5161 aaaaaaaaga atatataagg cttttaaagc ttttaaggtt taacggttgt ggacaacaag
     5221 ccagggatgt aacgcactga gaagccctta gagcctctca aagcaatttt gagtgacaca
     5281 ggaacactta acggctgaca tgggaattaa ttctgactct tatacacaag tgcggccctc
     5341 gaattgacgc gctccaccgc ggtggcggcc gctctagaac tagtggatcc cccgggctgc
     5401 aggaattcga tatccataaa ctgccaggaa ttggggatcg gattcgagct aagaattccc
     5461 ttggggtatc tttaaatact gtagaaaaga ggaaggaaat aataaatggc taaaatgaga
     5521 atatcaccgg aattgaaaaa actgatcgaa aaataccgct gcgtaaaaga tacggaagga
     5581 atgtctcctg ctaaggtata taagctggtg ggagaaaatg aaaacctata tttaaaaatg
     5641 acggacagcc ggtataaagg gaccacctat gatgtggaac gggaaaagga catgatgcta
     5701 tggctggaag gaaagctgcc tgttccaaag gtcctgcact ttgaacggca tgatggctgg
     5761 agcaatctgc tcatgagtga ggccgatggc gtcctttgct cggaagagta tgaagatgga
     5821 caaagccctg aaaagattat cgagctgtat gcggagtgca tcaggctctt tcactccatc
     5881 gacatatcgg attgtcccta tacgaatagc ttagacagcc gcttagccga attggattac
     5941 ttactgaata acgatctggc cgatgtggat tgcgaaaact gggaagaaga cactccattt
     6001 aaagatccgc gcgagctgta tgatttttta aagacggaaa agcccgaaga ggaacttgtc
     6061 ttttcccacg gcgacctggg agacagcaac atctttgtga aagatggcaa agtaagtggc
     6121 tttattgatc ttgggagaag cggcagggcg gacaagtggt atgacattgc cttctgcgtc
     6181 cggtcgatca gggaggatgt cggggaagaa cagtatgtcg agctattttt tgacttactg
     6241 gggatcaagc ctgattggga gaaaataaaa tattatattt tactggatga attgttttag
     6301 agctcgcacg ctgccgcaag cactcagggc gcaagggctg ctaaaggaag cggaacacgt
     6361 agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct
     6421 ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc
     6481 gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc
     6541 cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga
     6601 tctgatggcg caggggatca agatctgatc aagagacagg atactagtgg aggaagaaaa
     6661 a
//
