LOCUS       sequence-227072-        6634 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 6634)
  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 6634)
  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-Tn5-152
FEATURES             Location/Qualifiers
     source          1..6634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..3,4..516)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIE"
     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      701..795
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             839..1498
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(905..924)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1563..1649
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3393..3411)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3479..3583
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3584..3652,3653..4444)
                     /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(3802..3821)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            4683..4792
                     /note="incP origin of transfer"
     primer_bind     5346..5362
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5346..5362
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             5479..6273
                     /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 tgacctacgg 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 actacaacag ccacaacgtc 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 cgggtaatta gctgagcttg gactcctgtt gatagatcca
      721 gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt
      781 tttattggtg agaatccaag ctagcttggc gagattttca ggagctaagg aagctaaaat
      841 ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca
      901 ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat
      961 tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca
     1021 cattcttgcc cgcctgatga atgctcatcc ggaatttcgt atggcaatga aagacggtga
     1081 gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac
     1141 gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc
     1201 gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa
     1261 tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc
     1321 caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga
     1381 caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt
     1441 cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaatt
     1501 tttttaaggc agttattggt gcccttaaac gcctggggta atgactctct agcttgaggc
     1561 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     1621 cggtgaacgc tctcctgagt aggacaaatc cgccgctagg agcttgcggc cgccaggata
     1681 tcgacccaag taccgccacc taacaattcg ttcaagccga gatcggcttc ccggccgacg
     1741 cgtcaattcg agcgcgtcaa ttcgagggcc gcggcctagg cggccagatc tgatcaagag
     1801 acagtctagc tagaattaat tcgagtcgac cgatcccgta cacaagtagc gtcctgaacg
     1861 gaacctttcc cgttttccag gatctgatct tccatgtgac ctcctaacat ggtaacgttc
     1921 atgataactt ctgctcttca tcgtgcggcc gactgggcta aatctgtgtt ctcttcggcg
     1981 gcgctgggtg atcctcgccg tactgcccgc ttggttaacg tcgccgccca attggcaaaa
     2041 tattctggta aatcaataac catctcatca gagggtagtg aagccatgca ggaaggcgct
     2101 taccgattta tccgcaatcc caacgtttct gccgaggcga tcagaaaggc tggcgccatg
     2161 caaacagtca agttggctca ggagtttccc gaactgctgg ccattgagga caccacctct
     2221 ttgagttatc gccaccaggt cgccgaagag cttggcaagc tgggctctat tcaggataaa
     2281 tcccgcggat ggtgggttca ctccgttctc ttgctcgagg ccaccacatt ccgcaccgta
     2341 ggattactgc atcaggagtg gtggatgcgc ccggatgacc ctgccgatgc ggatgaaaag
     2401 gagagtggca aatggctggc agccgccgca actagccggt tacgcatggg cagcatgatg
     2461 agcaacgtga ttgcggtctg tgaccgcgaa gccgatattc atgcttatct gcaggacaaa
     2521 ctggcgcata acgagcgctt cgtggtgcgc tccaagcacc cacgcaagga cgtagagtct
     2581 gggttgtatc tgtacgacca tctgaagaac caaccggagt tgggtggcta tcagatcagc
     2641 attccgcaaa agggcgtggt ggataaacgc ggtaaacgta aaaatcgacc agcccgcaag
     2701 gcgagcttga gcctgcgcag tgggcgcatc acgctaaaac aggggaatat cacgctcaac
     2761 gcggtgctgg ccgaggagat taacccgccc aagggtgaga ccccgttgaa atggttgttg
     2821 ctgaccagcg aaccggtcga gtcgctagcc caagccttgc gcgtcatcga catttatacc
     2881 catcgctggc ggatcgagga gttccataag gcatggaaaa ccggagcagg agccgagagg
     2941 caacgcatgg aggagccgga taatctggag cggatggtct cgatcctctc gtttgttgcg
     3001 gtcaggctgt tacagctcag agaaagcttc acgctgccgc aagcactcag ggcgcaaggg
     3061 ctgctaaagg aagcggaaca cgtagaaagc cagtccgcag aaacggtgct gaccccggat
     3121 gaatgtcagc tactgggcta tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt
     3181 agcttgcagt gggcttacat ggcgatagct agactgggcg gttttatgga cagcaagcga
     3241 accggaattg ccagctgggg cgccctctgg gaaggttggg aagccctgca aagtaaactg
     3301 gatggctttc ttgccgccaa ggatctgatg gcgcagggga tcaagatctg atcggtcgat
     3361 cgaattaatt cccaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
     3421 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     3481 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3541 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3601 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3661 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3721 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3781 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     3841 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3901 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3961 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4021 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4081 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4141 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4201 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4261 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4321 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4381 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4441 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4501 atttaaaagg atctaggtga agatccgggg aattggggat caattccgtt ttccgctgca
     4561 taaccctgct tcggggtcat tatagcgatt ttttcggtat atccatcctt tttcgcacga
     4621 tatacaggat tttgccaaag ggttcgtgta gactttcctt ggtgtatcca acggcgtcag
     4681 ccgggcagga taggtgaagt aggcccaccc gcgagcgggt gttccttctt cactgtccct
     4741 tattcgcacc tggcggtgct caacgggaat cctgctctgc gaggctggcc ggctaccgcc
     4801 ggcgtaacag atgagggcaa gcggatggct gatgaaacca agccaaccag gaagggcagc
     4861 ccacctatca cggaattgat ccggccacga tgcgtccggc gtagaggatc tgaagatcag
     4921 cagttcaacc tgttgatagt acgtactaag ctctcatgtt tcacgtacta agctctcatg
     4981 tttaacgtac taagctctca tgtttaacga actaaaccct catggctaac gtactaagct
     5041 ctcatgtttc acgtactaag ctctcatgtt tgaacaataa aattaatata aatcagcaac
     5101 ttaaatagcc tctaaggttt taagttttat aagaaaaaaa agaatatata aggcttttaa
     5161 agcttttaag gtttaacggt tgtggacaac aagccaggga tgtaacgcac tgagaagccc
     5221 ttagagcctc tcaaagcaat tttgagtgac acaggaacac ttaacggctg acatgggaat
     5281 taattctgac tcttatacac aagtgcggcc ctcgaattga cgcgctccac cgcggtggcg
     5341 gccgctctag aactagtgga tcccccgggc tgcaggaatt cgatatccat aaactgccag
     5401 gaattgggga tcggattcga gctaagaatt cccttggggt atctttaaat actgtagaaa
     5461 agaggaagga aataataaat ggctaaaatg agaatatcac cggaattgaa aaaactgatc
     5521 gaaaaatacc gctgcgtaaa agatacggaa ggaatgtctc ctgctaaggt atataagctg
     5581 gtgggagaaa atgaaaacct atatttaaaa atgacggaca gccggtataa agggaccacc
     5641 tatgatgtgg aacgggaaaa ggacatgatg ctatggctgg aaggaaagct gcctgttcca
     5701 aaggtcctgc actttgaacg gcatgatggc tggagcaatc tgctcatgag tgaggccgat
     5761 ggcgtccttt gctcggaaga gtatgaagat ggacaaagcc ctgaaaagat tatcgagctg
     5821 tatgcggagt gcatcaggct ctttcactcc atcgacatat cggattgtcc ctatacgaat
     5881 agcttagaca gccgcttagc cgaattggat tacttactga ataacgatct ggccgatgtg
     5941 gattgcgaaa actgggaaga agacactcca tttaaagatc cgcgcgagct gtatgatttt
     6001 ttaaagacgg aaaagcccga agaggaactt gtcttttccc acggcgacct gggagacagc
     6061 aacatctttg tgaaagatgg caaagtaagt ggctttattg atcttgggag aagcggcagg
     6121 gcggacaagt ggtatgacat tgccttctgc gtccggtcga tcagggagga tgtcggggaa
     6181 gaacagtatg tcgagctatt ttttgactta ctggggatca agcctgattg ggagaaaata
     6241 aaatattata ttttactgga tgaattgttt tagagctcgc acgctgccgc aagcactcag
     6301 ggcgcaaggg ctgctaaagg aagcggaaca cgtagaaagc cagtccgcag aaacggtgct
     6361 gaccccggat gaatgtcagc tactgggcta tctggacaag ggaaaacgca agcgcaaaga
     6421 gaaagcaggt agcttgcagt gggcttacat ggcgatagct agactgggcg gttttatgga
     6481 cagcaagcga accggaattg ccagctgggg cgccctctgg taaggttggg aagccctgca
     6541 aagtaaactg gatggctttc ttgccgccaa ggatctgatg gcgcagggga tcaagatctg
     6601 atcaagagac aggatactag tggaggaaga aaaa
//
