LOCUS       sequence-227220-        6671 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 6671)
  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 6671)
  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-Tn5-153
FEATURES             Location/Qualifiers
     source          1..6671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..3,4..6,7..717)
                     /codon_start=1
                     /product="monomeric optimized variant of EYFP, identical to
                     mVenus but with L68V (Kremers et al., 2006)"
                     /label=SYFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(46..67)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(307..326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     654..675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      738..832
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             876..1535
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(942..961)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1600..1686
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3430..3448)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3516..3620
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3621..3689,3690..4481)
                     /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(3839..3858)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            4720..4829
                     /note="incP origin of transfer"
     primer_bind     5383..5399
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5383..5399
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             5516..6310
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     GQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDVGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
       61 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      121 ggcaagctga ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc
      181 ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag
      241 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      301 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      361 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      421 aagctggagt acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac
      481 ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
      541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      601 tacctgagct accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      661 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
      721 gtaattagct gagcttggac tcctgttgat agatccagta atgacctcag aactccatct
      781 ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagcta
      841 gcttggcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata
      901 ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg
      961 ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa
     1021 agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg
     1081 ctcatccgga atttcgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc
     1141 acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat
     1201 accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg
     1261 aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc
     1321 cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc
     1381 ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga
     1441 ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac
     1501 aacagtactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt tattggtgcc
     1561 cttaaacgcc tggggtaatg actctctagc ttgaggcatc aaataaaacg aaaggctcag
     1621 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     1681 acaaatccgc cgctaggagc ttgcggccgc caggatatcg acccaagtac cgccacctaa
     1741 caattcgttc aagccgagat cggcttcccg gccgacgcgt caattcgagc gcgtcaattc
     1801 gagggccgcg gcctaggcgg ccagatctga tcaagagaca gtctagctag aattaattcg
     1861 agtcgaccga tcccgtacac aagtagcgtc ctgaacggaa cctttcccgt tttccaggat
     1921 ctgatcttcc atgtgacctc ctaacatggt aacgttcatg ataacttctg ctcttcatcg
     1981 tgcggccgac tgggctaaat ctgtgttctc ttcggcggcg ctgggtgatc ctcgccgtac
     2041 tgcccgcttg gttaacgtcg ccgcccaatt ggcaaaatat tctggtaaat caataaccat
     2101 ctcatcagag ggtagtgaag ccatgcagga aggcgcttac cgatttatcc gcaatcccaa
     2161 cgtttctgcc gaggcgatca gaaaggctgg cgccatgcaa acagtcaagt tggctcagga
     2221 gtttcccgaa ctgctggcca ttgaggacac cacctctttg agttatcgcc accaggtcgc
     2281 cgaagagctt ggcaagctgg gctctattca ggataaatcc cgcggatggt gggttcactc
     2341 cgttctcttg ctcgaggcca ccacattccg caccgtagga ttactgcatc aggagtggtg
     2401 gatgcgcccg gatgaccctg ccgatgcgga tgaaaaggag agtggcaaat ggctggcagc
     2461 cgccgcaact agccggttac gcatgggcag catgatgagc aacgtgattg cggtctgtga
     2521 ccgcgaagcc gatattcatg cttatctgca ggacaaactg gcgcataacg agcgcttcgt
     2581 ggtgcgctcc aagcacccac gcaaggacgt agagtctggg ttgtatctgt acgaccatct
     2641 gaagaaccaa ccggagttgg gtggctatca gatcagcatt ccgcaaaagg gcgtggtgga
     2701 taaacgcggt aaacgtaaaa atcgaccagc ccgcaaggcg agcttgagcc tgcgcagtgg
     2761 gcgcatcacg ctaaaacagg ggaatatcac gctcaacgcg gtgctggccg aggagattaa
     2821 cccgcccaag ggtgagaccc cgttgaaatg gttgttgctg accagcgaac cggtcgagtc
     2881 gctagcccaa gccttgcgcg tcatcgacat ttatacccat cgctggcgga tcgaggagtt
     2941 ccataaggca tggaaaaccg gagcaggagc cgagaggcaa cgcatggagg agccggataa
     3001 tctggagcgg atggtctcga tcctctcgtt tgttgcggtc aggctgttac agctcagaga
     3061 aagcttcacg ctgccgcaag cactcagggc gcaagggctg ctaaaggaag cggaacacgt
     3121 agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct
     3181 ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc
     3241 gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc
     3301 cctctgggaa ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga
     3361 tctgatggcg caggggatca agatctgatc ggtcgatcga attaattccc aattcttgaa
     3421 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     3481 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3541 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3601 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     3661 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3721 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3781 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     3841 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     3901 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     3961 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4021 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4081 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     4141 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     4201 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     4261 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     4321 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     4381 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     4441 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     4501 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     4561 tccggggaat tggggatcaa ttccgttttc cgctgcataa ccctgcttcg gggtcattat
     4621 agcgattttt tcggtatatc catccttttt cgcacgatat acaggatttt gccaaagggt
     4681 tcgtgtagac tttccttggt gtatccaacg gcgtcagccg ggcaggatag gtgaagtagg
     4741 cccacccgcg agcgggtgtt ccttcttcac tgtcccttat tcgcacctgg cggtgctcaa
     4801 cgggaatcct gctctgcgag gctggccggc taccgccggc gtaacagatg agggcaagcg
     4861 gatggctgat gaaaccaagc caaccaggaa gggcagccca cctatcacgg aattgatccg
     4921 gccacgatgc gtccggcgta gaggatctga agatcagcag ttcaacctgt tgatagtacg
     4981 tactaagctc tcatgtttca cgtactaagc tctcatgttt aacgtactaa gctctcatgt
     5041 ttaacgaact aaaccctcat ggctaacgta ctaagctctc atgtttcacg tactaagctc
     5101 tcatgtttga acaataaaat taatataaat cagcaactta aatagcctct aaggttttaa
     5161 gttttataag aaaaaaaaga atatataagg cttttaaagc ttttaaggtt taacggttgt
     5221 ggacaacaag ccagggatgt aacgcactga gaagccctta gagcctctca aagcaatttt
     5281 gagtgacaca ggaacactta acggctgaca tgggaattaa ttctgactct tatacacaag
     5341 tgcggccctc gaattgacgc gctccaccgc ggtggcggcc gctctagaac tagtggatcc
     5401 cccgggctgc aggaattcga tatccataaa ctgccaggaa ttggggatcg gattcgagct
     5461 aagaattccc ttggggtatc tttaaatact gtagaaaaga ggaaggaaat aataaatggc
     5521 taaaatgaga atatcaccgg aattgaaaaa actgatcgaa aaataccgct gcgtaaaaga
     5581 tacggaagga atgtctcctg ctaaggtata taagctggtg ggagaaaatg aaaacctata
     5641 tttaaaaatg acggacagcc ggtataaagg gaccacctat gatgtggaac gggaaaagga
     5701 catgatgcta tggctggaag gaaagctgcc tgttccaaag gtcctgcact ttgaacggca
     5761 tgatggctgg agcaatctgc tcatgagtga ggccgatggc gtcctttgct cggaagagta
     5821 tgaagatgga caaagccctg aaaagattat cgagctgtat gcggagtgca tcaggctctt
     5881 tcactccatc gacatatcgg attgtcccta tacgaatagc ttagacagcc gcttagccga
     5941 attggattac ttactgaata acgatctggc cgatgtggat tgcgaaaact gggaagaaga
     6001 cactccattt aaagatccgc gcgagctgta tgatttttta aagacggaaa agcccgaaga
     6061 ggaacttgtc ttttcccacg gcgacctggg agacagcaac atctttgtga aagatggcaa
     6121 agtaagtggc tttattgatc ttgggagaag cggcagggcg gacaagtggt atgacattgc
     6181 cttctgcgtc cggtcgatca gggaggatgt cggggaagaa cagtatgtcg agctattttt
     6241 tgacttactg gggatcaagc ctgattggga gaaaataaaa tattatattt tactggatga
     6301 attgttttag agctcgcacg ctgccgcaag cactcagggc gcaagggctg ctaaaggaag
     6361 cggaacacgt agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac
     6421 tgggctatct ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg
     6481 cttacatggc gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca
     6541 gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg
     6601 ccgccaagga tctgatggcg caggggatca agatctgatc aagagacagg atactagtgg
     6661 aggaagaaaa a
//
