LOCUS       sequence-227061-        6625 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 6625)
  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 6625)
  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-Tn5-141
FEATURES             Location/Qualifiers
     source          1..6625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(673..675,676..1389)
                     /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(715..736)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(976..995)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1323..1344
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1403..1497
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1541..2200
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1607..1626)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2265..2351
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(4095..4113)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4181..4285
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4286..4354,4355..5146)
                     /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(4504..4523)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            5385..5494
                     /note="incP origin of transfer"
     primer_bind     6048..6064
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6048..6064
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        join(6185..6190,6191..6207,6208..6213)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             join(6402..6625,1..310)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSSEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 attcatcgcg cttgctgcct tcgaccaaga agcggttgtt ggcgctctcg cggcttacgt
       61 tctgcccaag tttgagcagc cgcgtagtga gatctatatc tatgatctcg cagtctccag
      121 cgagcaccgg aggcagggca ttgccaccgc gctcatcaat ctcctcaagc atgaggccaa
      181 cgcgcttggt gcttatgtga tctacgtgca agcagattac ggtgacgatc ccgcagtggc
      241 tctctataca aagttgggca tacgggaaga agtgatgcac tttgatatcg acccaagtac
      301 cgccacctaa agctcgcacg ctgccgcaag cactcagggc gcaagggctg ctaaaggaag
      361 cggaacacgt agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac
      421 tgggctatct ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg
      481 cttacatggc gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca
      541 gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg
      601 ccgccaagga tctgatggcg caggggatca agatctgatc aagagacagg atactagtgg
      661 aggaagaaaa aaatgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
      721 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
      781 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
      841 tggcccaccc tcgtgaccac cctgtcctgg ggcgtgcagt gcttcgcccg ctaccccgac
      901 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
      961 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     1021 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     1081 ctggggcaca agctggagta caactacttt agcgacaacg tctatatcac cgccgacaag
     1141 cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg
     1201 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1261 gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga gaagcgcgat
     1321 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     1381 tacaagtaat tagctgagct tggactcctg ttgatagatc cagtaatgac ctcagaactc
     1441 catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg tgagaatcca
     1501 agctagcttg gcgagatttt caggagctaa ggaagctaaa atggagaaaa aaatcactgg
     1561 atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg catttcagtc
     1621 agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct ttttaaagac
     1681 cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg cccgcctgat
     1741 gaatgctcat ccggaatttc gtatggcaat gaaagacggt gagctggtga tatgggatag
     1801 tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat cgctctggag
     1861 tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg tggcgtgtta
     1921 cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt tcgtctcagc
     1981 caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg acaacttctt
     2041 cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct
     2101 ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa tgcttaatga
     2161 attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag gcagttattg
     2221 gtgcccttaa acgcctgggg taatgactct ctagcttgag gcatcaaata aaacgaaagg
     2281 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
     2341 gtaggacaaa tccgccgcta ggagcttgcg gccgccagga tatcgaccca agtaccgcca
     2401 cctaacaatt cgttcaagcc gagatcggct tcccggccga cgcgtcaatt cgagcgcgtc
     2461 aattcgaggg ccgcggccta ggcggccaga tctgatcaag agacagtcta gctagaatta
     2521 attcgagtcg accgatcccg tacacaagta gcgtcctgaa cggaaccttt cccgttttcc
     2581 aggatctgat cttccatgtg acctcctaac atggtaacgt tcatgataac ttctgctctt
     2641 catcgtgcgg ccgactgggc taaatctgtg ttctcttcgg cggcgctggg tgatcctcgc
     2701 cgtactgccc gcttggttaa cgtcgccgcc caattggcaa aatattctgg taaatcaata
     2761 accatctcat cagagggtag tgaagccatg caggaaggcg cttaccgatt tatccgcaat
     2821 cccaacgttt ctgccgaggc gatcagaaag gctggcgcca tgcaaacagt caagttggct
     2881 caggagtttc ccgaactgct ggccattgag gacaccacct ctttgagtta tcgccaccag
     2941 gtcgccgaag agcttggcaa gctgggctct attcaggata aatcccgcgg atggtgggtt
     3001 cactccgttc tcttgctcga ggccaccaca ttccgcaccg taggattact gcatcaggag
     3061 tggtggatgc gcccggatga ccctgccgat gcggatgaaa aggagagtgg caaatggctg
     3121 gcagccgccg caactagccg gttacgcatg ggcagcatga tgagcaacgt gattgcggtc
     3181 tgtgaccgcg aagccgatat tcatgcttat ctgcaggaca aactggcgca taacgagcgc
     3241 ttcgtggtgc gctccaagca cccacgcaag gacgtagagt ctgggttgta tctgtacgac
     3301 catctgaaga accaaccgga gttgggtggc tatcagatca gcattccgca aaagggcgtg
     3361 gtggataaac gcggtaaacg taaaaatcga ccagcccgca aggcgagctt gagcctgcgc
     3421 agtgggcgca tcacgctaaa acaggggaat atcacgctca acgcggtgct ggccgaggag
     3481 attaacccgc ccaagggtga gaccccgttg aaatggttgt tgctgaccag cgaaccggtc
     3541 gagtcgctag cccaagcctt gcgcgtcatc gacatttata cccatcgctg gcggatcgag
     3601 gagttccata aggcatggaa aaccggagca ggagccgaga ggcaacgcat ggaggagccg
     3661 gataatctgg agcggatggt ctcgatcctc tcgtttgttg cggtcaggct gttacagctc
     3721 agagaaagct tcacgctgcc gcaagcactc agggcgcaag ggctgctaaa ggaagcggaa
     3781 cacgtagaaa gccagtccgc agaaacggtg ctgaccccgg atgaatgtca gctactgggc
     3841 tatctggaca agggaaaacg caagcgcaaa gagaaagcag gtagcttgca gtgggcttac
     3901 atggcgatag ctagactggg cggttttatg gacagcaagc gaaccggaat tgccagctgg
     3961 ggcgccctct gggaaggttg ggaagccctg caaagtaaac tggatggctt tcttgccgcc
     4021 aaggatctga tggcgcaggg gatcaagatc tgatcggtcg atcgaattaa ttcccaattc
     4081 ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4141 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4201 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4261 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4321 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4381 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4441 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4501 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4561 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4621 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4681 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4741 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4801 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4861 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4921 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4981 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5041 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5101 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5161 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5221 gaagatccgg ggaattgggg atcaattccg ttttccgctg cataaccctg cttcggggtc
     5281 attatagcga ttttttcggt atatccatcc tttttcgcac gatatacagg attttgccaa
     5341 agggttcgtg tagactttcc ttggtgtatc caacggcgtc agccgggcag gataggtgaa
     5401 gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca cctggcggtg
     5461 ctcaacggga atcctgctct gcgaggctgg ccggctaccg ccggcgtaac agatgagggc
     5521 aagcggatgg ctgatgaaac caagccaacc aggaagggca gcccacctat cacggaattg
     5581 atccggccac gatgcgtccg gcgtagagga tctgaagatc agcagttcaa cctgttgata
     5641 gtacgtacta agctctcatg tttcacgtac taagctctca tgtttaacgt actaagctct
     5701 catgtttaac gaactaaacc ctcatggcta acgtactaag ctctcatgtt tcacgtacta
     5761 agctctcatg tttgaacaat aaaattaata taaatcagca acttaaatag cctctaaggt
     5821 tttaagtttt ataagaaaaa aaagaatata taaggctttt aaagctttta aggtttaacg
     5881 gttgtggaca acaagccagg gatgtaacgc actgagaagc ccttagagcc tctcaaagca
     5941 attttgagtg acacaggaac acttaacggc tgacatggga attaattctg actcttatac
     6001 acaagtgcgg ccctcgaatt gacgcgctcc accgcggtgg cggccgctct agaactagtg
     6061 gatcccccgg gctgcaggaa ttcgatatcc ataaactgcc aggaattggg gatcggattc
     6121 gagctttagt acctagattt agatgtctaa aaagcggggt accgaggacg cgtcaattct
     6181 cgaattgaca taagcctgtt cggttcgtaa actgtaatgc aagtagcgta tgcgctcacg
     6241 caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat
     6301 ggcttgttat gactgttttt ttgtacagtc tatgcctcgg gcatccaagc agcaagcgcg
     6361 ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagcaacg
     6421 atgttacgca gcagggcagt cgccctaaaa caaagttagg tggctcaagt atgggcatca
     6481 ttcgcacatg taggctcggc cctgaccaag tcaaatccat gcgggctgct cttgatcttt
     6541 tcggtcgtga gttcggagac gtagccacct actcccaaca tcagccggac tccgattacc
     6601 tcgggaactt gctccgtagt aagac
//
