LOCUS       sequence-226605-        6665 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 6665)
  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 6665)
  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-Tn5-151
FEATURES             Location/Qualifiers
     source          1..6665
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..3,4..717)
                     /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(43..64)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(304..323)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     651..672
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      731..825
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             869..1528
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(935..954)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1593..1679
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3423..3441)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3509..3613
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3614..3682,3683..4474)
                     /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(3832..3851)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            4713..4822
                     /note="incP origin of transfer"
     primer_bind     5376..5392
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5376..5392
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             5509..6303
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     GQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDVGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 atgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
       61 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      121 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      181 gtgaccaccc tgtcctgggg cgtgcagtgc ttcgcccgct accccgacca catgaagcag
      241 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
      301 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
      361 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
      421 ctggagtaca actactttag cgacaacgtc tatatcaccg ccgacaagca gaagaacggc
      481 atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac
      541 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
      601 ctgagcaccc agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg
      661 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaatta
      721 gctgagcttg gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg
      781 ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag ctagcttggc
      841 gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt
      901 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg
      961 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa
     1021 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc
     1081 ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg
     1141 ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga
     1201 cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct
     1261 ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt
     1321 gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt
     1381 caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt
     1441 tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta
     1501 ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattggt gcccttaaac
     1561 gcctggggta atgactctct agcttgaggc atcaaataaa acgaaaggct cagtcgaaag
     1621 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
     1681 cgccgctagg agcttgcggc cgccaggata tcgacccaag taccgccacc taacaattcg
     1741 ttcaagccga gatcggcttc ccggccgacg cgtcaattcg agcgcgtcaa ttcgagggcc
     1801 gcggcctagg cggccagatc tgatcaagag acagtctagc tagaattaat tcgagtcgac
     1861 cgatcccgta cacaagtagc gtcctgaacg gaacctttcc cgttttccag gatctgatct
     1921 tccatgtgac ctcctaacat ggtaacgttc atgataactt ctgctcttca tcgtgcggcc
     1981 gactgggcta aatctgtgtt ctcttcggcg gcgctgggtg atcctcgccg tactgcccgc
     2041 ttggttaacg tcgccgccca attggcaaaa tattctggta aatcaataac catctcatca
     2101 gagggtagtg aagccatgca ggaaggcgct taccgattta tccgcaatcc caacgtttct
     2161 gccgaggcga tcagaaaggc tggcgccatg caaacagtca agttggctca ggagtttccc
     2221 gaactgctgg ccattgagga caccacctct ttgagttatc gccaccaggt cgccgaagag
     2281 cttggcaagc tgggctctat tcaggataaa tcccgcggat ggtgggttca ctccgttctc
     2341 ttgctcgagg ccaccacatt ccgcaccgta ggattactgc atcaggagtg gtggatgcgc
     2401 ccggatgacc ctgccgatgc ggatgaaaag gagagtggca aatggctggc agccgccgca
     2461 actagccggt tacgcatggg cagcatgatg agcaacgtga ttgcggtctg tgaccgcgaa
     2521 gccgatattc atgcttatct gcaggacaaa ctggcgcata acgagcgctt cgtggtgcgc
     2581 tccaagcacc cacgcaagga cgtagagtct gggttgtatc tgtacgacca tctgaagaac
     2641 caaccggagt tgggtggcta tcagatcagc attccgcaaa agggcgtggt ggataaacgc
     2701 ggtaaacgta aaaatcgacc agcccgcaag gcgagcttga gcctgcgcag tgggcgcatc
     2761 acgctaaaac aggggaatat cacgctcaac gcggtgctgg ccgaggagat taacccgccc
     2821 aagggtgaga ccccgttgaa atggttgttg ctgaccagcg aaccggtcga gtcgctagcc
     2881 caagccttgc gcgtcatcga catttatacc catcgctggc ggatcgagga gttccataag
     2941 gcatggaaaa ccggagcagg agccgagagg caacgcatgg aggagccgga taatctggag
     3001 cggatggtct cgatcctctc gtttgttgcg gtcaggctgt tacagctcag agaaagcttc
     3061 acgctgccgc aagcactcag ggcgcaaggg ctgctaaagg aagcggaaca cgtagaaagc
     3121 cagtccgcag aaacggtgct gaccccggat gaatgtcagc tactgggcta tctggacaag
     3181 ggaaaacgca agcgcaaaga gaaagcaggt agcttgcagt gggcttacat ggcgatagct
     3241 agactgggcg gttttatgga cagcaagcga accggaattg ccagctgggg cgccctctgg
     3301 gaaggttggg aagccctgca aagtaaactg gatggctttc ttgccgccaa ggatctgatg
     3361 gcgcagggga tcaagatctg atcggtcgat cgaattaatt cccaattctt gaagacgaaa
     3421 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     3481 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3541 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3601 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3661 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3721 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3781 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3841 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     3901 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3961 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4021 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4081 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4141 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4201 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4261 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4321 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4381 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4441 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4501 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatccgggg
     4561 aattggggat caattccgtt ttccgctgca taaccctgct tcggggtcat tatagcgatt
     4621 ttttcggtat atccatcctt tttcgcacga tatacaggat tttgccaaag ggttcgtgta
     4681 gactttcctt ggtgtatcca acggcgtcag ccgggcagga taggtgaagt aggcccaccc
     4741 gcgagcgggt gttccttctt cactgtccct tattcgcacc tggcggtgct caacgggaat
     4801 cctgctctgc gaggctggcc ggctaccgcc ggcgtaacag atgagggcaa gcggatggct
     4861 gatgaaacca agccaaccag gaagggcagc ccacctatca cggaattgat ccggccacga
     4921 tgcgtccggc gtagaggatc tgaagatcag cagttcaacc tgttgatagt acgtactaag
     4981 ctctcatgtt tcacgtacta agctctcatg tttaacgtac taagctctca tgtttaacga
     5041 actaaaccct catggctaac gtactaagct ctcatgtttc acgtactaag ctctcatgtt
     5101 tgaacaataa aattaatata aatcagcaac ttaaatagcc tctaaggttt taagttttat
     5161 aagaaaaaaa agaatatata aggcttttaa agcttttaag gtttaacggt tgtggacaac
     5221 aagccaggga tgtaacgcac tgagaagccc ttagagcctc tcaaagcaat tttgagtgac
     5281 acaggaacac ttaacggctg acatgggaat taattctgac tcttatacac aagtgcggcc
     5341 ctcgaattga cgcgctccac cgcggtggcg gccgctctag aactagtgga tcccccgggc
     5401 tgcaggaatt cgatatccat aaactgccag gaattgggga tcggattcga gctaagaatt
     5461 cccttggggt atctttaaat actgtagaaa agaggaagga aataataaat ggctaaaatg
     5521 agaatatcac cggaattgaa aaaactgatc gaaaaatacc gctgcgtaaa agatacggaa
     5581 ggaatgtctc ctgctaaggt atataagctg gtgggagaaa atgaaaacct atatttaaaa
     5641 atgacggaca gccggtataa agggaccacc tatgatgtgg aacgggaaaa ggacatgatg
     5701 ctatggctgg aaggaaagct gcctgttcca aaggtcctgc actttgaacg gcatgatggc
     5761 tggagcaatc tgctcatgag tgaggccgat ggcgtccttt gctcggaaga gtatgaagat
     5821 ggacaaagcc ctgaaaagat tatcgagctg tatgcggagt gcatcaggct ctttcactcc
     5881 atcgacatat cggattgtcc ctatacgaat agcttagaca gccgcttagc cgaattggat
     5941 tacttactga ataacgatct ggccgatgtg gattgcgaaa actgggaaga agacactcca
     6001 tttaaagatc cgcgcgagct gtatgatttt ttaaagacgg aaaagcccga agaggaactt
     6061 gtcttttccc acggcgacct gggagacagc aacatctttg tgaaagatgg caaagtaagt
     6121 ggctttattg atcttgggag aagcggcagg gcggacaagt ggtatgacat tgccttctgc
     6181 gtccggtcga tcagggagga tgtcggggaa gaacagtatg tcgagctatt ttttgactta
     6241 ctggggatca agcctgattg ggagaaaata aaatattata ttttactgga tgaattgttt
     6301 tagagctcgc acgctgccgc aagcactcag ggcgcaaggg ctgctaaagg aagcggaaca
     6361 cgtagaaagc cagtccgcag aaacggtgct gaccccggat gaatgtcagc tactgggcta
     6421 tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt agcttgcagt gggcttacat
     6481 ggcgatagct agactgggcg gttttatgga cagcaagcga accggaattg ccagctgggg
     6541 cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc ttgccgccaa
     6601 ggatctgatg gcgcagggga tcaagatctg atcaagagac aggatactag tggaggaaga
     6661 aaaaa
//
