LOCUS       sequence-366484-        6630 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 6630)
  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 6630)
  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-Tn5-142
FEATURES             Location/Qualifiers
     source          1..6630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            152..261
                     /note="incP origin of transfer"
     primer_bind     815..831
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     815..831
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        join(952..957,958..974,975..980)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1169..1702
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     CDS             join(2064..2066,2067..2069,2070..2783)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(2109..2130)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2717..2738
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      2800..2894
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2938..3597
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3004..3023)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3662..3748
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(5492..5510)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5578..5682
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5683..5751,5752..6543)
                     /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(5901..5920)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 attggggatc aattccgttt tccgctgcat aaccctgctt cggggtcatt atagcgattt
       61 tttcggtata tccatccttt ttcgcacgat atacaggatt ttgccaaagg gttcgtgtag
      121 actttccttg gtgtatccaa cggcgtcagc cgggcaggat aggtgaagta ggcccacccg
      181 cgagcgggtg ttccttcttc actgtccctt attcgcacct ggcggtgctc aacgggaatc
      241 ctgctctgcg aggctggccg gctaccgccg gcgtaacaga tgagggcaag cggatggctg
      301 atgaaaccaa gccaaccagg aagggcagcc cacctatcac ggaattgatc cggccacgat
      361 gcgtccggcg tagaggatct gaagatcagc agttcaacct gttgatagta cgtactaagc
      421 tctcatgttt cacgtactaa gctctcatgt ttaacgtact aagctctcat gtttaacgaa
      481 ctaaaccctc atggctaacg tactaagctc tcatgtttca cgtactaagc tctcatgttt
      541 gaacaataaa attaatataa atcagcaact taaatagcct ctaaggtttt aagttttata
      601 agaaaaaaaa gaatatataa ggcttttaaa gcttttaagg tttaacggtt gtggacaaca
      661 agccagggat gtaacgcact gagaagccct tagagcctct caaagcaatt ttgagtgaca
      721 caggaacact taacggctga catgggaatt aattctgact cttatacaca agtgcggccc
      781 tcgaattgac gcgctccacc gcggtggcgg ccgctctaga actagtggat cccccgggct
      841 gcaggaattc gatatccata aactgccagg aattggggat cggattcgag ctttagtacc
      901 tagatttaga tgtctaaaaa gcggggtacc gaggacgcgt caattctcga attgacataa
      961 gcctgttcgg ttcgtaaact gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga
     1021 accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac
     1081 tgtttttttg tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt
     1141 cgatgtttga tgttatggag cagcaacgat gttacgcagc agcaacgatg ttacgcagca
     1201 gggcagtcgc cctaaaacaa agttaggtgg ctcaagtatg ggcatcattc gcacatgtag
     1261 gctcggccct gaccaagtca aatccatgcg ggctgctctt gatcttttcg gtcgtgagtt
     1321 cggagacgta gccacctact cccaacatca gccggactcc gattacctcg ggaacttgct
     1381 ccgtagtaag acattcatcg cgcttgctgc cttcgaccaa gaagcggttg ttggcgctct
     1441 cgcggcttac gttctgccca agtttgagca gccgcgtagt gagatctata tctatgatct
     1501 cgcagtctcc ggcgagcacc ggaggcaggg cattgccacc gcgctcatca atctcctcaa
     1561 gcatgaggcc aacgcgcttg gtgcttatgt gatctacgtg caagcagatt acggtgacga
     1621 tcccgcagtg gctctctata caaagttggg catacgggaa gaagtgatgc actttgatat
     1681 cgacccaagt accgccacct aaagctcgca cgctgccgca agcactcagg gcgcaagggc
     1741 tgctaaagga agcggaacac gtagaaagcc agtccgcaga aacggtgctg accccggatg
     1801 aatgtcagct actgggctat ctggacaagg gaaaacgcaa gcgcaaagag aaagcaggta
     1861 gcttgcagtg ggcttacatg gcgatagcta gactgggcgg ttttatggac agcaagcgaa
     1921 ccggaattgc cagctggggc gccctctggt aaggttggga agccctgcaa agtaaactgg
     1981 atggctttct tgccgccaag gatctgatgg cgcaggggat caagatctga tcaagagaca
     2041 ggatactagt ggaggaagaa aaaatggtga gcaagggcga ggagctgttc accggggtgg
     2101 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtccgcggcg
     2161 agggcgaggg cgatgccacc aacggcaagc tgaccctgaa gttcatctgc accaccggca
     2221 agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcgtg gcctgcttca
     2281 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2341 acgtccagga gcgcaccatc tctttcaagg acgacggtac ctacaagacc cgcgccgagg
     2401 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2461 aggacggcaa catcctgggg cacaagctgg agtacaactt caacagccac tacgtctata
     2521 tcacggccga caagcagaag aactgcatca aggctaactt caagatccgc cacaacgttg
     2581 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2641 ccgtgctgct gcccgacaac cactacctga gccatcagtc caagctgagc aaagacccca
     2701 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg attacacatg
     2761 gcatggacga gctgtacaag taataattag ctgagcttgg actcctgttg atagatccag
     2821 taatgacctc agaactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt
     2881 ttattggtga gaatccaagc tagcttggcg agattttcag gagctaagga agctaaaatg
     2941 gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat
     3001 tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt
     3061 acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac
     3121 attcttgccc gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa agacggtgag
     3181 ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg
     3241 ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca catatattcg
     3301 caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat
     3361 atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc
     3421 aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac
     3481 aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc
     3541 ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt
     3601 ttttaaggca gttattggtg cccttaaacg cctggggtaa tgactctcta gcttgaggca
     3661 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     3721 ggtgaacgct ctcctgagta ggacaaatcc gccgctagga gcttgcggcc gccaggatat
     3781 cgacccaagt accgccacct aacaattcgt tcaagccgag atcggcttcc cggccgacgc
     3841 gtcaattcga gcgcgtcaat tcgagggccg cggcctaggc ggccagatct gatcaagaga
     3901 cagtctagct agaattaatt cgagtcgacc gatcccgtac acaagtagcg tcctgaacgg
     3961 aacctttccc gttttccagg atctgatctt ccatgtgacc tcctaacatg gtaacgttca
     4021 tgataacttc tgctcttcat cgtgcggccg actgggctaa atctgtgttc tcttcggcgg
     4081 cgctgggtga tcctcgccgt actgcccgct tggttaacgt cgccgcccaa ttggcaaaat
     4141 attctggtaa atcaataacc atctcatcag agggtagtga agccatgcag gaaggcgctt
     4201 accgatttat ccgcaatccc aacgtttctg ccgaggcgat cagaaaggct ggcgccatgc
     4261 aaacagtcaa gttggctcag gagtttcccg aactgctggc cattgaggac accacctctt
     4321 tgagttatcg ccaccaggtc gccgaagagc ttggcaagct gggctctatt caggataaat
     4381 cccgcggatg gtgggttcac tccgttctct tgctcgaggc caccacattc cgcaccgtag
     4441 gattactgca tcaggagtgg tggatgcgcc cggatgaccc tgccgatgcg gatgaaaagg
     4501 agagtggcaa atggctggca gccgccgcaa ctagccggtt acgcatgggc agcatgatga
     4561 gcaacgtgat tgcggtctgt gaccgcgaag ccgatattca tgcttatctg caggacaaac
     4621 tggcgcataa cgagcgcttc gtggtgcgct ccaagcaccc acgcaaggac gtagagtctg
     4681 ggttgtatct gtacgaccat ctgaagaacc aaccggagtt gggtggctat cagatcagca
     4741 ttccgcaaaa gggcgtggtg gataaacgcg gtaaacgtaa aaatcgacca gcccgcaagg
     4801 cgagcttgag cctgcgcagt gggcgcatca cgctaaaaca ggggaatatc acgctcaacg
     4861 cggtgctggc cgaggagatt aacccgccca agggtgagac cccgttgaaa tggttgttgc
     4921 tgaccagcga accggtcgag tcgctagccc aagccttgcg cgtcatcgac atttataccc
     4981 atcgctggcg gatcgaggag ttccataagg catggaaaac cggagcagga gccgagaggc
     5041 aacgcatgga ggagccggat aatctggagc ggatggtctc gatcctctcg tttgttgcgg
     5101 tcaggctgtt acagctcaga gaaagcttca cgctgccgca agcactcagg gcgcaagggc
     5161 tgctaaagga agcggaacac gtagaaagcc agtccgcaga aacggtgctg accccggatg
     5221 aatgtcagct actgggctat ctggacaagg gaaaacgcaa gcgcaaagag aaagcaggta
     5281 gcttgcagtg ggcttacatg gcgatagcta gactgggcgg ttttatggac agcaagcgaa
     5341 ccggaattgc cagctggggc gccctctggg aaggttggga agccctgcaa agtaaactgg
     5401 atggctttct tgccgccaag gatctgatgg cgcaggggat caagatctga tcggtcgatc
     5461 gaattaattc ccaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     5521 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     5581 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5641 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5701 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5761 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5821 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5881 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5941 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     6001 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     6061 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     6121 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     6181 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     6241 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     6301 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     6361 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     6421 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     6481 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     6541 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     6601 tttaaaagga tctaggtgaa gatccgggga
//
