LOCUS       sequence-227099-        7776 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 7776)
  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 7776)
  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-Tn5-161
FEATURES             Location/Qualifiers
     source          1..7776
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     193..214
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      273..367
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             411..1070
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(477..496)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1135..1221
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2965..2983)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3051..3155
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3156..3224,3225..4016)
                     /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(3374..3393)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            4255..4364
                     /note="incP origin of transfer"
     primer_bind     4918..4934
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4918..4934
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             complement(4997..6196)
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     CDS             join(7319..7321,7322..7776,1..259)
                     /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(7361..7382)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7622..7641)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga
       61 cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
      121 gctgctgccc gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga
      181 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
      241 ggacgagctg tacaagtaat tagctgagct tggactcctg ttgatagatc cagtaatgac
      301 ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg
      361 tgagaatcca agctagcttg gcgagatttt caggagctaa ggaagctaaa atggagaaaa
      421 aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg
      481 catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct
      541 ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg
      601 cccgcctgat gaatgctcat ccggaatttc gtatggcaat gaaagacggt gagctggtga
      661 tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat
      721 cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg
      781 tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt
      841 tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg
      901 acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc
      961 tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa
     1021 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag
     1081 gcagttattg gtgcccttaa acgcctgggg taatgactct ctagcttgag gcatcaaata
     1141 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     1201 gctctcctga gtaggacaaa tccgccgcta ggagcttgcg gccgccagga tatcgaccca
     1261 agtaccgcca cctaacaatt cgttcaagcc gagatcggct tcccggccga cgcgtcaatt
     1321 cgagcgcgtc aattcgaggg ccgcggccta ggcggccaga tctgatcaag agacagtcta
     1381 gctagaatta attcgagtcg accgatcccg tacacaagta gcgtcctgaa cggaaccttt
     1441 cccgttttcc aggatctgat cttccatgtg acctcctaac atggtaacgt tcatgataac
     1501 ttctgctctt catcgtgcgg ccgactgggc taaatctgtg ttctcttcgg cggcgctggg
     1561 tgatcctcgc cgtactgccc gcttggttaa cgtcgccgcc caattggcaa aatattctgg
     1621 taaatcaata accatctcat cagagggtag tgaagccatg caggaaggcg cttaccgatt
     1681 tatccgcaat cccaacgttt ctgccgaggc gatcagaaag gctggcgcca tgcaaacagt
     1741 caagttggct caggagtttc ccgaactgct ggccattgag gacaccacct ctttgagtta
     1801 tcgccaccag gtcgccgaag agcttggcaa gctgggctct attcaggata aatcccgcgg
     1861 atggtgggtt cactccgttc tcttgctcga ggccaccaca ttccgcaccg taggattact
     1921 gcatcaggag tggtggatgc gcccggatga ccctgccgat gcggatgaaa aggagagtgg
     1981 caaatggctg gcagccgccg caactagccg gttacgcatg ggcagcatga tgagcaacgt
     2041 gattgcggtc tgtgaccgcg aagccgatat tcatgcttat ctgcaggaca aactggcgca
     2101 taacgagcgc ttcgtggtgc gctccaagca cccacgcaag gacgtagagt ctgggttgta
     2161 tctgtacgac catctgaaga accaaccgga gttgggtggc tatcagatca gcattccgca
     2221 aaagggcgtg gtggataaac gcggtaaacg taaaaatcga ccagcccgca aggcgagctt
     2281 gagcctgcgc agtgggcgca tcacgctaaa acaggggaat atcacgctca acgcggtgct
     2341 ggccgaggag attaacccgc ccaagggtga gaccccgttg aaatggttgt tgctgaccag
     2401 cgaaccggtc gagtcgctag cccaagcctt gcgcgtcatc gacatttata cccatcgctg
     2461 gcggatcgag gagttccata aggcatggaa aaccggagca ggagccgaga ggcaacgcat
     2521 ggaggagccg gataatctgg agcggatggt ctcgatcctc tcgtttgttg cggtcaggct
     2581 gttacagctc agagaaagct tcacgctgcc gcaagcactc agggcgcaag ggctgctaaa
     2641 ggaagcggaa cacgtagaaa gccagtccgc agaaacggtg ctgaccccgg atgaatgtca
     2701 gctactgggc tatctggaca agggaaaacg caagcgcaaa gagaaagcag gtagcttgca
     2761 gtgggcttac atggcgatag ctagactggg cggttttatg gacagcaagc gaaccggaat
     2821 tgccagctgg ggcgccctct gggaaggttg ggaagccctg caaagtaaac tggatggctt
     2881 tcttgccgcc aaggatctga tggcgcaggg gatcaagatc tgatcggtcg atcgaattaa
     2941 ttcccaattc ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     3001 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3061 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3121 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3181 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3241 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3301 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3361 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     3421 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3481 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3541 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3601 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3661 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     3721 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3781 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3841 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3901 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3961 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4021 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4081 ggatctaggt gaagatccgg ggaattgggg atcaattccg ttttccgctg cataaccctg
     4141 cttcggggtc attatagcga ttttttcggt atatccatcc tttttcgcac gatatacagg
     4201 attttgccaa agggttcgtg tagactttcc ttggtgtatc caacggcgtc agccgggcag
     4261 gataggtgaa gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca
     4321 cctggcggtg ctcaacggga atcctgctct gcgaggctgg ccggctaccg ccggcgtaac
     4381 agatgagggc aagcggatgg ctgatgaaac caagccaacc aggaagggca gcccacctat
     4441 cacggaattg atccggccac gatgcgtccg gcgtagagga tctgaagatc agcagttcaa
     4501 cctgttgata gtacgtacta agctctcatg tttcacgtac taagctctca tgtttaacgt
     4561 actaagctct catgtttaac gaactaaacc ctcatggcta acgtactaag ctctcatgtt
     4621 tcacgtacta agctctcatg tttgaacaat aaaattaata taaatcagca acttaaatag
     4681 cctctaaggt tttaagtttt ataagaaaaa aaagaatata taaggctttt aaagctttta
     4741 aggtttaacg gttgtggaca acaagccagg gatgtaacgc actgagaagc ccttagagcc
     4801 tctcaaagca attttgagtg acacaggaac acttaacggc tgacatggga attaattctg
     4861 actcttatac acaagtgcgg ccctcgaatt gacgcgctcc accgcggtgg cggccgctct
     4921 agaactagtg gatcccccgg gctgcaggaa ttcgatatcc ataaactgcc aggaattggg
     4981 gatcggattc gagctctcag cgatcggctc gttgccctgc gccgctccaa agcccgcgac
     5041 gcagcgccgg caggcagagc aagtagaggg cagcgcctgc aatccatgcc cacccgttcc
     5101 acgttgttat agaagccgca tagatcgccg tgaagaggag gggtccgacg atcgaggtca
     5161 ggctggtgag cgccgccagt gagccttgca gctgcccctg acgttcctca tccacctgcc
     5221 tggacaacat tgcttgcagc gccggcattc cgatgccacc cgaagcaagc aggaccatga
     5281 tcgggaacgc catccatccc cgtgtcgcga aggcaagcag gatgtagcct gtgccgtcgg
     5341 caatcattcc gagcatgagt gcccgccttt cgccgagccg ggcggctaca gggccggtga
     5401 tcattgcctg ggcgagtgaa tgcagaatgc caaatgcggc aagcgaaatg ccgatcgtgg
     5461 tcgcgtccca gtgaaagcga tcctcgccga aaatgaccca aagcgcggcc ggcacctgtc
     5521 cgacaagttg catgatgaag aagaccgcca tcagggcggc gacgacggtc atgccccggg
     5581 cccaccggaa cgaagcgagc gggttgagag cctcccggcg taacggccgg cgttcgcctt
     5641 tgtgcgactc cggcaaaagg aaacagcccg tcaggaaatt gaggccgttc aaggctgccg
     5701 cggcgaagaa cggagcgtgg ggggagaaac cgcccatcag cccaccgagc acaggtcccg
     5761 cgaccatccc gaacccgaaa caggcgctca tgaagccgaa gtgccgcgcg cgctcatcgc
     5821 catcagtgat atcggcaata taagcgccgg ctaccgcccc agtcgccccg gtgatgccgg
     5881 ccacgatccg cccgatatag agaacccaaa ggaaaggcgc tgtcgccatg atggcgtagt
     5941 cgacagtggc gccggccagc gagacgagca agattggccg ccgcccgaaa cgatccgaca
     6001 gcgcgcccag cacaggtgcg caggcaaatt gcaccaacgc atacagcgcc agcagaatgc
     6061 catagtgggc ggtgacgtcg ttcgagtgaa ccagatcgcg caggaggccc ggcagcaccg
     6121 gcataatcag gccgatgccg acagcgtcga gcgcgacagt gctcagaatt acgatcaggg
     6181 gtatgttggg tttcacgtct ggcctccgga ccagccgatt gaacgcgcgg attctttatc
     6241 actgataagt tggtggacat attatgttta tcagtgataa agtgtcaagc atgacaaagt
     6301 tgcagccgaa tacagtgatc cgtgccgccc tggacctgtt gaacgaggtc ggcgtagacg
     6361 gtctgacgac acgcaaactg gcggaacggt tgggggttca gcagccggcg ctttactggc
     6421 acttcaggaa caagcgggcg ctgctcgacg cactggccga agccatgctg gcggagaatc
     6481 atacgcattc ggtgccgaga gccgacgacg actggcgctc atttctgatc gggaatgccc
     6541 gcagcttcag gcaggcgctg ctcgcctacc gcgatggcgc gcgcatccat gccggcacgc
     6601 gaccgggcgc accgcagatg gaaacggccg acgcgcagct tcgcttcctc tgcgaggcgg
     6661 gtttttcggc cggggacgcc gtcaatgcgc tgatgacaat cagctacttc actgttgggg
     6721 ccgtgcttga ggagcaggcc ggcgacagcg atgccggcga gcgcggcggc accgttgaac
     6781 aggctccgct ctcgccgctg ttgcgggccg cgatagacgc cttcgacgaa gccggtccgg
     6841 acgcagcgtt cgagcaggga ctcgcggtga ttgtcgatgg attggcgaaa aggaggctcg
     6901 ttgtcaggaa cgttgaagga ccgagaaagg gtgctaagaa accattatta tcatgaagct
     6961 cgcacgctgc cgcaagcact cagggcgcaa gggctgctaa aggaagcgga acacgtagaa
     7021 agccagtccg cagaaacggt gctgaccccg gatgaatgtc agctactggg ctatctggac
     7081 aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc agtgggctta catggcgata
     7141 gctagactgg gcggttttat ggacagcaag cgaaccggaa ttgccagctg gggcgccctc
     7201 tggtaaggtt gggaagccct gcaaagtaaa ctggatggct ttcttgccgc caaggatctg
     7261 atggcgcagg ggatcaagat ctgatcaaga gacaggatac tagtggagga agaaaaaaat
     7321 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     7381 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     7441 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     7501 gaccaccctg tcctggggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
     7561 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     7621 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     7681 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     7741 ggagtacaac tactttagcg acaacgtcta tatcac
//
