LOCUS       sequence-227083-        7783 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 7783)
  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 7783)
  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-Tn5-164
FEATURES             Location/Qualifiers
     source          1..7783
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      634..728
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             772..1431
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(838..857)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1496..1582
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3326..3344)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3412..3516
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3517..3585,3586..4377)
                     /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(3735..3754)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            4616..4725
                     /note="incP origin of transfer"
     primer_bind     5279..5295
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5279..5295
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             complement(5358..6557)
                     /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(7690..7783,1..617)
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
ORIGIN
        1 tcgagatcga gggcgagggc gagggccgcc cctacgaggg ctttcagacc gctaagctga
       61 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct catttcacct
      121 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttc aagctgtcct
      181 tccccgaggg cttcaagtgg gagcgcgtga tgaactacga ggacggcggc gtggtgaccg
      241 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
      301 ccaacttccc ctccgacggc cccgtgatgc agaagaagac catgggctgg gaggcctcct
      361 ccgagcggat gtaccccgag gacggtgccc tgaagggcaa gatcaagatg aggctgaagc
      421 tgaaggacgg cggccactac acctccgagg tcaagaccac ctacaaggcc aagaagcccg
      481 tgcagctgcc cggcgcctac atcgtcgaca tcaagttgga catcacctcc cacaacgagg
      541 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
      601 acgagctgta caagtaataa ttagctgagc ttggactcct gttgatagat ccagtaatga
      661 cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg
      721 gtgagaatcc aagctagctt ggcgagattt tcaggagcta aggaagctaa aatggagaaa
      781 aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag
      841 gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc
      901 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt
      961 gcccgcctga tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg
     1021 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca
     1081 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat
     1141 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt
     1201 ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg
     1261 gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg
     1321 ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga
     1381 atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta atttttttaa
     1441 ggcagttatt ggtgccctta aacgcctggg gtaatgactc tctagcttga ggcatcaaat
     1501 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     1561 cgctctcctg agtaggacaa atccgccgct aggagcttgc ggccgccagg atatcgaccc
     1621 aagtaccgcc acctaacaat tcgttcaagc cgagatcggc ttcccggccg acgcgtcaat
     1681 tcgagcgcgt caattcgagg gccgcggcct aggcggccag atctgatcaa gagacagtct
     1741 agctagaatt aattcgagtc gaccgatccc gtacacaagt agcgtcctga acggaacctt
     1801 tcccgttttc caggatctga tcttccatgt gacctcctaa catggtaacg ttcatgataa
     1861 cttctgctct tcatcgtgcg gccgactggg ctaaatctgt gttctcttcg gcggcgctgg
     1921 gtgatcctcg ccgtactgcc cgcttggtta acgtcgccgc ccaattggca aaatattctg
     1981 gtaaatcaat aaccatctca tcagagggta gtgaagccat gcaggaaggc gcttaccgat
     2041 ttatccgcaa tcccaacgtt tctgccgagg cgatcagaaa ggctggcgcc atgcaaacag
     2101 tcaagttggc tcaggagttt cccgaactgc tggccattga ggacaccacc tctttgagtt
     2161 atcgccacca ggtcgccgaa gagcttggca agctgggctc tattcaggat aaatcccgcg
     2221 gatggtgggt tcactccgtt ctcttgctcg aggccaccac attccgcacc gtaggattac
     2281 tgcatcagga gtggtggatg cgcccggatg accctgccga tgcggatgaa aaggagagtg
     2341 gcaaatggct ggcagccgcc gcaactagcc ggttacgcat gggcagcatg atgagcaacg
     2401 tgattgcggt ctgtgaccgc gaagccgata ttcatgctta tctgcaggac aaactggcgc
     2461 ataacgagcg cttcgtggtg cgctccaagc acccacgcaa ggacgtagag tctgggttgt
     2521 atctgtacga ccatctgaag aaccaaccgg agttgggtgg ctatcagatc agcattccgc
     2581 aaaagggcgt ggtggataaa cgcggtaaac gtaaaaatcg accagcccgc aaggcgagct
     2641 tgagcctgcg cagtgggcgc atcacgctaa aacaggggaa tatcacgctc aacgcggtgc
     2701 tggccgagga gattaacccg cccaagggtg agaccccgtt gaaatggttg ttgctgacca
     2761 gcgaaccggt cgagtcgcta gcccaagcct tgcgcgtcat cgacatttat acccatcgct
     2821 ggcggatcga ggagttccat aaggcatgga aaaccggagc aggagccgag aggcaacgca
     2881 tggaggagcc ggataatctg gagcggatgg tctcgatcct ctcgtttgtt gcggtcaggc
     2941 tgttacagct cagagaaagc ttcacgctgc cgcaagcact cagggcgcaa gggctgctaa
     3001 aggaagcgga acacgtagaa agccagtccg cagaaacggt gctgaccccg gatgaatgtc
     3061 agctactggg ctatctggac aagggaaaac gcaagcgcaa agagaaagca ggtagcttgc
     3121 agtgggctta catggcgata gctagactgg gcggttttat ggacagcaag cgaaccggaa
     3181 ttgccagctg gggcgccctc tgggaaggtt gggaagccct gcaaagtaaa ctggatggct
     3241 ttcttgccgc caaggatctg atggcgcagg ggatcaagat ctgatcggtc gatcgaatta
     3301 attcccaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     3361 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3421 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3481 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     3541 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     3601 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     3661 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     3721 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     3781 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     3841 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     3901 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     3961 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4021 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     4081 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4141 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4201 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     4261 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     4321 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     4381 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     4441 aggatctagg tgaagatccg gggaattggg gatcaattcc gttttccgct gcataaccct
     4501 gcttcggggt cattatagcg attttttcgg tatatccatc ctttttcgca cgatatacag
     4561 gattttgcca aagggttcgt gtagactttc cttggtgtat ccaacggcgt cagccgggca
     4621 ggataggtga agtaggccca cccgcgagcg ggtgttcctt cttcactgtc ccttattcgc
     4681 acctggcggt gctcaacggg aatcctgctc tgcgaggctg gccggctacc gccggcgtaa
     4741 cagatgaggg caagcggatg gctgatgaaa ccaagccaac caggaagggc agcccaccta
     4801 tcacggaatt gatccggcca cgatgcgtcc ggcgtagagg atctgaagat cagcagttca
     4861 acctgttgat agtacgtact aagctctcat gtttcacgta ctaagctctc atgtttaacg
     4921 tactaagctc tcatgtttaa cgaactaaac cctcatggct aacgtactaa gctctcatgt
     4981 ttcacgtact aagctctcat gtttgaacaa taaaattaat ataaatcagc aacttaaata
     5041 gcctctaagg ttttaagttt tataagaaaa aaaagaatat ataaggcttt taaagctttt
     5101 aaggtttaac ggttgtggac aacaagccag ggatgtaacg cactgagaag cccttagagc
     5161 ctctcaaagc aattttgagt gacacaggaa cacttaacgg ctgacatggg aattaattct
     5221 gactcttata cacaagtgcg gccctcgaat tgacgcgctc caccgcggtg gcggccgctc
     5281 tagaactagt ggatcccccg ggctgcagga attcgatatc cataaactgc caggaattgg
     5341 ggatcggatt cgagctctca gcgatcggct cgttgccctg cgccgctcca aagcccgcga
     5401 cgcagcgccg gcaggcagag caagtagagg gcagcgcctg caatccatgc ccacccgttc
     5461 cacgttgtta tagaagccgc atagatcgcc gtgaagagga ggggtccgac gatcgaggtc
     5521 aggctggtga gcgccgccag tgagccttgc agctgcccct gacgttcctc atccacctgc
     5581 ctggacaaca ttgcttgcag cgccggcatt ccgatgccac ccgaagcaag caggaccatg
     5641 atcgggaacg ccatccatcc ccgtgtcgcg aaggcaagca ggatgtagcc tgtgccgtcg
     5701 gcaatcattc cgagcatgag tgcccgcctt tcgccgagcc gggcggctac agggccggtg
     5761 atcattgcct gggcgagtga atgcagaatg ccaaatgcgg caagcgaaat gccgatcgtg
     5821 gtcgcgtccc agtgaaagcg atcctcgccg aaaatgaccc aaagcgcggc cggcacctgt
     5881 ccgacaagtt gcatgatgaa gaagaccgcc atcagggcgg cgacgacggt catgccccgg
     5941 gcccaccgga acgaagcgag cgggttgaga gcctcccggc gtaacggccg gcgttcgcct
     6001 ttgtgcgact ccggcaaaag gaaacagccc gtcaggaaat tgaggccgtt caaggctgcc
     6061 gcggcgaaga acggagcgtg gggggagaaa ccgcccatca gcccaccgag cacaggtccc
     6121 gcgaccatcc cgaacccgaa acaggcgctc atgaagccga agtgccgcgc gcgctcatcg
     6181 ccatcagtga tatcggcaat ataagcgccg gctaccgccc cagtcgcccc ggtgatgccg
     6241 gccacgatcc gcccgatata gagaacccaa aggaaaggcg ctgtcgccat gatggcgtag
     6301 tcgacagtgg cgccggccag cgagacgagc aagattggcc gccgcccgaa acgatccgac
     6361 agcgcgccca gcacaggtgc gcaggcaaat tgcaccaacg catacagcgc cagcagaatg
     6421 ccatagtggg cggtgacgtc gttcgagtga accagatcgc gcaggaggcc cggcagcacc
     6481 ggcataatca ggccgatgcc gacagcgtcg agcgcgacag tgctcagaat tacgatcagg
     6541 ggtatgttgg gtttcacgtc tggcctccgg accagcctcc gctggtccga ttgaacgcgc
     6601 ggattcttta tcactgataa gttggtggac atattatgtt tatcagtgat aaagtttcaa
     6661 gcatgacaaa gttgcagccg aatacagtga tccgtgccgc cctggacctg ttgaacgagg
     6721 tcggcgtaga cggtctgacg acacgcaaac tggcggaacg gttgggggtt cagcagccgg
     6781 cgctttactg gcacttcagg aacaagcggg cgctgctcga cgcactggcc gaagccatgc
     6841 tggcggagaa tcatacgcat tcggtgccga gagccgacga cgactggcgc tcatttctga
     6901 tcgggaatgc ccgcagcttc aggcaggcgc tgctcgccta ccgcgatggc gcgcgcatcc
     6961 atgccggcac gcgaccgggc gcaccgcaga tggaaacggc cgacgcgcag cttcgcttcc
     7021 tctgcgaggc gggtttttcg gccggggacg ccgtcaatgc gctgatgaca atcagctact
     7081 tcactgttgg ggccgtgctt gaggagcagg ccggcgacag cgatgccggc gagcgcggcg
     7141 gcaccgttga acaggctccg ctctcgccgc tgttgcgggc cgcgatagac gccttcgacg
     7201 aagccggtcc ggacgcagcg ttcgagcagg gactcgcggt gattgtcgat ggattggcga
     7261 aaaggaggct cgttgtcagg aacgttgaag gaccgagaaa gggtgctaag aaaccattat
     7321 tatcatgaag ctcgcacgct gccgcaagca ctcagggcgc aagggctgct aaaggaagcg
     7381 gaacacgtag aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg tcagctactg
     7441 ggctatctgg acaagggaaa acgcaagcgc aaagagaaag caggtagctt gcagtgggct
     7501 tacatggcga tagctagact gggcggtttt atggacagca agcgaaccgg aattgccagc
     7561 tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc
     7621 gccaaggatc tgatggcgca ggggatcaag atctgatcaa gagacaggat actagtggag
     7681 gaagaaaaaa tggtgagcaa gggcgaggag aataacatgg ccatcatcaa ggagttcatg
     7741 cgcttcaagg tgcgcatgga gggctccgtg aacggccacg agt
//
