LOCUS       sequence-227213-        6635 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 6635)
  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 6635)
  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-Tn5-156
FEATURES             Location/Qualifiers
     source          1..6635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      909..1003
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1047..1706
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1113..1132)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1771..1857
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3601..3619)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3687..3791
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3792..3860,3861..4652)
                     /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(4010..4029)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            4891..5000
                     /note="incP origin of transfer"
     primer_bind     5554..5570
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5554..5570
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             5687..6481
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     GQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDVGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 ggtagcttgc agtgggctta catggcgata gctagactgg gcggttttat ggacagcaag
       61 cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt gggaatccct gcaaagtaaa
      121 ctggatggct ttcttgccgc caaggatctg atggcgcagg ggatcaagat ctgatcaaga
      181 gacaggatac tagtggagga agaaaaaatg gtgagcaagg gcgaggagct gatcaaggag
      241 aacatgcaca tgaagctgta catggaaggc accgtgaaca accaccactt caagtgcacc
      301 accgaagggg agggcaagcc ctacgagggc acccagaccc agaggattaa ggtggtggag
      361 ggaggccccc tgccgttcgc attcgacatc ctggccacct gctttatgta cgggagcaag
      421 accttcatca accacaccca gggcatcccc gatttcttta agcagtcctt ccctgagggc
      481 ttcacatggg agagagtcac cacatacgaa gacgggggcg tgcttaccgt tacccaggac
      541 accagcctcc aggacggctg cttgatctac aacgtcaagc tcagaggggt gaacttccca
      601 tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg aggccaccac cgagaccctg
      661 taccccgctg acggcggcct ggaaggcaga tgcgacatgg ccctgaagct cgtgggcggg
      721 ggccacctgc actgcaacct gaagaccaca tacagatcca agaaacccgc taagaacctc
      781 aagatgcccg gcgtctactt tgtggaccgc agactggaaa gaatcaagga ggccgacaat
      841 gagacctacg tcgagcagca cgaggtggct gtggccagat actgcgacct cctaattagc
      901 tgagcttgga ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt
      961 cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagct agcttggcga
     1021 gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg
     1081 atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta
     1141 cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata
     1201 agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg
     1261 aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt
     1321 acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg
     1381 atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg
     1441 cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga
     1501 gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca
     1561 ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc
     1621 atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact
     1681 gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc
     1741 ctggggtaat gactctctag cttgaggcat caaataaaac gaaaggctca gtcgaaagac
     1801 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     1861 ccgctaggag cttgcggccg ccaggatatc gacccaagta ccgccaccta acaattcgtt
     1921 caagccgaga tcggcttccc ggccgacgcg tcaattcgag cgcgtcaatt cgagggccgc
     1981 ggcctaggcg gccagatctg atcaagagac agtctagcta gaattaattc gagtcgaccg
     2041 atcccgtaca caagtagcgt cctgaacgga acctttcccg ttttccagga tctgatcttc
     2101 catgtgacct cctaacatgg taacgttcat gataacttct gctcttcatc gtgcggccga
     2161 ctgggctaaa tctgtgttct cttcggcggc gctgggtgat cctcgccgta ctgcccgctt
     2221 ggttaacgtc gccgcccaat tggcaaaata ttctggtaaa tcaataacca tctcatcaga
     2281 gggtagtgaa gccatgcagg aaggcgctta ccgatttatc cgcaatccca acgtttctgc
     2341 cgaggcgatc agaaaggctg gcgccatgca aacagtcaag ttggctcagg agtttcccga
     2401 actgctggcc attgaggaca ccacctcttt gagttatcgc caccaggtcg ccgaagagct
     2461 tggcaagctg ggctctattc aggataaatc ccgcggatgg tgggttcact ccgttctctt
     2521 gctcgaggcc accacattcc gcaccgtagg attactgcat caggagtggt ggatgcgccc
     2581 ggatgaccct gccgatgcgg atgaaaagga gagtggcaaa tggctggcag ccgccgcaac
     2641 tagccggtta cgcatgggca gcatgatgag caacgtgatt gcggtctgtg accgcgaagc
     2701 cgatattcat gcttatctgc aggacaaact ggcgcataac gagcgcttcg tggtgcgctc
     2761 caagcaccca cgcaaggacg tagagtctgg gttgtatctg tacgaccatc tgaagaacca
     2821 accggagttg ggtggctatc agatcagcat tccgcaaaag ggcgtggtgg ataaacgcgg
     2881 taaacgtaaa aatcgaccag cccgcaaggc gagcttgagc ctgcgcagtg ggcgcatcac
     2941 gctaaaacag gggaatatca cgctcaacgc ggtgctggcc gaggagatta acccgcccaa
     3001 gggtgagacc ccgttgaaat ggttgttgct gaccagcgaa ccggtcgagt cgctagccca
     3061 agccttgcgc gtcatcgaca tttataccca tcgctggcgg atcgaggagt tccataaggc
     3121 atggaaaacc ggagcaggag ccgagaggca acgcatggag gagccggata atctggagcg
     3181 gatggtctcg atcctctcgt ttgttgcggt caggctgtta cagctcagag aaagcttcac
     3241 gctgccgcaa gcactcaggg cgcaagggct gctaaaggaa gcggaacacg tagaaagcca
     3301 gtccgcagaa acggtgctga ccccggatga atgtcagcta ctgggctatc tggacaaggg
     3361 aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag
     3421 actgggcggt tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggga
     3481 aggttgggaa gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc
     3541 gcaggggatc aagatctgat cggtcgatcg aattaattcc caattcttga agacgaaagg
     3601 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     3661 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3721 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3781 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3841 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3901 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3961 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4021 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4081 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4141 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4201 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4261 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4321 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4381 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4441 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4501 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4561 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4621 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4681 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atccggggaa
     4741 ttggggatca attccgtttt ccgctgcata accctgcttc ggggtcatta tagcgatttt
     4801 ttcggtatat ccatcctttt tcgcacgata tacaggattt tgccaaaggg ttcgtgtaga
     4861 ctttccttgg tgtatccaac ggcgtcagcc gggcaggata ggtgaagtag gcccacccgc
     4921 gagcgggtgt tccttcttca ctgtccctta ttcgcacctg gcggtgctca acgggaatcc
     4981 tgctctgcga ggctggccgg ctaccgccgg cgtaacagat gagggcaagc ggatggctga
     5041 tgaaaccaag ccaaccagga agggcagccc acctatcacg gaattgatcc ggccacgatg
     5101 cgtccggcgt agaggatctg aagatcagca gttcaacctg ttgatagtac gtactaagct
     5161 ctcatgtttc acgtactaag ctctcatgtt taacgtacta agctctcatg tttaacgaac
     5221 taaaccctca tggctaacgt actaagctct catgtttcac gtactaagct ctcatgtttg
     5281 aacaataaaa ttaatataaa tcagcaactt aaatagcctc taaggtttta agttttataa
     5341 gaaaaaaaag aatatataag gcttttaaag cttttaaggt ttaacggttg tggacaacaa
     5401 gccagggatg taacgcactg agaagccctt agagcctctc aaagcaattt tgagtgacac
     5461 aggaacactt aacggctgac atgggaatta attctgactc ttatacacaa gtgcggccct
     5521 cgaattgacg cgctccaccg cggtggcggc cgctctagaa ctagtggatc ccccgggctg
     5581 caggaattcg atatccataa actgccagga attggggatc ggattcgagc taagaattcc
     5641 cttggggtat ctttaaatac tgtagaaaag aggaaggaaa taataaatgg ctaaaatgag
     5701 aatatcaccg gaattgaaaa aactgatcga aaaataccgc tgcgtaaaag atacggaagg
     5761 aatgtctcct gctaaggtat ataagctggt gggagaaaat gaaaacctat atttaaaaat
     5821 gacggacagc cggtataaag ggaccaccta tgatgtggaa cgggaaaagg acatgatgct
     5881 atggctggaa ggaaagctgc ctgttccaaa ggtcctgcac tttgaacggc atgatggctg
     5941 gagcaatctg ctcatgagtg aggccgatgg cgtcctttgc tcggaagagt atgaagatgg
     6001 acaaagccct gaaaagatta tcgagctgta tgcggagtgc atcaggctct ttcactccat
     6061 cgacatatcg gattgtccct atacgaatag cttagacagc cgcttagccg aattggatta
     6121 cttactgaat aacgatctgg ccgatgtgga ttgcgaaaac tgggaagaag acactccatt
     6181 taaagatccg cgcgagctgt atgatttttt aaagacggaa aagcccgaag aggaacttgt
     6241 cttttcccac ggcgacctgg gagacagcaa catctttgtg aaagatggca aagtaagtgg
     6301 ctttattgat cttgggagaa gcggcagggc ggacaagtgg tatgacattg ccttctgcgt
     6361 ccggtcgatc agggaggatg tcggggaaga acagtatgtc gagctatttt ttgacttact
     6421 ggggatcaag cctgattggg agaaaataaa atattatatt ttactggatg aattgtttta
     6481 gagctcgcac gctgccgcaa gcactcaggg cgcaagggct gctaaaggaa gcggaacacg
     6541 tagaaagcca gtccgcagaa acggtgctga ccccggatga atgtcagcta ctgggctatc
     6601 tggacaaggg aaaacgcaag cgcaaagaga aagca
//
