LOCUS       sequence-368333-        7756 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 7756)
  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 7756)
  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-Tn5-162
FEATURES             Location/Qualifiers
     source          1..7756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            72..181
                     /note="incP origin of transfer"
     primer_bind     735..751
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     735..751
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             complement(814..2013)
                     /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(3146..3148,3149..3661)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIE"
     primer_bind     complement(3188..3209)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3449..3468)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3796..3817
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      3846..3940
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3984..4643
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4050..4069)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      4708..4794
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(6538..6556)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6624..6728
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6729..6797,6798..7589)
                     /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(6947..6966)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttcgcacgat atacaggatt ttgccaaagg gttcgtgtag actttccttg gtgtatccaa
       61 cggcgtcagc cgggcaggat aggtgaagta ggcccacccg cgagcgggtg ttccttcttc
      121 actgtccctt attcgcacct ggcggtgctc aacgggaatc ctgctctgcg aggctggccg
      181 gctaccgccg gcgtaacaga tgagggcaag cggatggctg atgaaaccaa gccaaccagg
      241 aagggcagcc cacctatcac ggaattgatc cggccacgat gcgtccggcg tagaggatct
      301 gaagatcagc agttcaacct gttgatagta cgtactaagc tctcatgttt cacgtactaa
      361 gctctcatgt ttaacgtact aagctctcat gtttaacgaa ctaaaccctc atggctaacg
      421 tactaagctc tcatgtttca cgtactaagc tctcatgttt gaacaataaa attaatataa
      481 atcagcaact taaatagcct ctaaggtttt aagttttata agaaaaaaaa gaatatataa
      541 ggcttttaaa gcttttaagg tttaacggtt gtggacaaca agccagggat gtaacgcact
      601 gagaagccct tagagcctct caaagcaatt ttgagtgaca caggaacact taacggctga
      661 catgggaatt aattctgact cttatacaca agtgcggccc tcgaattgac gcgctccacc
      721 gcggtggcgg ccgctctaga actagtggat cccccgggct gcaggaattc gatatccata
      781 aactgccagg aattggggat cggattcgag ctctcagcga tcggctcgtt gccctgcgcc
      841 gctccaaagc ccgcgacgca gcgccggcag gcagagcaag tagagggcag cgcctgcaat
      901 ccatgcccac ccgttccacg ttgttataga agccgcatag atcgccgtga agaggagggg
      961 tccgacgatc gaggtcaggc tggtgagcgc cgccagtgag ccttgcagct gcccctgacg
     1021 ttcctcatcc acctgcctgg acaacattgc ttgcagcgcc ggcattccga tgccacccga
     1081 agcaagcagg accatgatcg ggaacgccat ccatccccgt gtcgcgaagg caagcaggat
     1141 gtagcctgtg ccgtcggcaa tcattccgag catgagtgcc cgcctttcgc cgagccgggc
     1201 ggctacaggg ccggtgatca ttgcctgggc gagtgaatgc agaatgccaa atgcggcaag
     1261 cgaaatgccg atcgtggtcg cgtcccagtg aaagcgatcc tcgccgaaaa tgacccaaag
     1321 cgcggccggc acctgtccga caagttgcat gatgaagaag accgccatca gggcggcgac
     1381 gacggtcatg ccccgggccc accggaacga agcgagcggg ttgagagcct cccggcgtaa
     1441 cggccggcgt tcgcctttgt gcgactccgg caaaaggaaa cagcccgtca ggaaattgag
     1501 gccgttcaag gctgccgcgg cgaagaacgg agcgtggggg gagaaaccgc ccatcagccc
     1561 accgagcaca ggtcccgcga ccatcccgaa cccgaaacag gcgctcatga agccgaagtg
     1621 ccgcgcgcgc tcatcgccat cagtgatatc ggcaatataa gcgccggcta ccgccccagt
     1681 cgccccggtg atgccggcca cgatccgccc gatatagaga acccaaagga aaggcgctgt
     1741 cgccatgatg gcgtagtcga cagtggcgcc ggccagcgag acgagcaaga ttggccgccg
     1801 cccgaaacga tccgacagcg cgcccagcac aggtgcgcag gcaaattgca ccaacgcata
     1861 cagcgccagc agaatgccat agtgggcggt gacgtcgttc gagtgaacca gatcgcgcag
     1921 gaggcccggc agcaccggca taatcaggcc gatgccgaca gcgtcgagcg cgacagtgct
     1981 cagaattacg atcaggggta tgttgggttt cacgtctggc ctccggacca gcctctgctg
     2041 gtccgattga acgcgcggat tctttatcac tgataagttg gtggacatat tatgtttatc
     2101 agtgataaag tgtcaagcat gacaaagttg cagccgaata cagtgatccg tgccgccctg
     2161 gacctgttga acgaggtcgg cgtagacggt ctgacgacac gcaaactggc ggaacggttg
     2221 ggggttcagc agccggcgct ttactggcac ttcaggaaca agcgggcgct gctcgacgca
     2281 ctggccgaag ccatgctggc ggagaatcat acgcattcgg tgccgagagc cgacgacgac
     2341 tggcgctcat ttctgatcgg gaatgcccgc agcttcaggc aggcgctgct cgcctaccgc
     2401 gatggcgcgc gcatccatgc cggcacgcga ccgggcgcac cgcagatgga aacggccgac
     2461 gcgcagcttc gcttcctctg cgaggcgggt ttttcggccg gggacgccgt caatgcgctg
     2521 atgacaatca gctacttcac tgttggggcc gtgcttgagg agcaggccgg cgacagcgat
     2581 gccggcgagc gcggcggcac cgttgaacag gctccgctct cgccgctgtt gcgggccgcg
     2641 atagactcct tcgacgaagc cggtccggac gcagcgttcg agcagggact cgcggtgatt
     2701 gtcgatggat tggcgaaaag gaggctcgtt gtcaggaacg ttgaaggacc gagaaagggt
     2761 gctaagaaac cattattatc atgaagctcg cacgctgccg caagcactca gggcgcaagg
     2821 gctgctaaag gaagcggaac acgtagaaag ccagtccgca gaaacggtgc tgaccccgga
     2881 tgaatgtcag ctactgggct atctggacaa gggaaaacgc aagcgcaaag agaaagcagg
     2941 tagcttgcag tgggcttaca tggcgatagc tagactgggc ggttttatgg acagcaagcg
     3001 aaccggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc aaagtaaact
     3061 ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagatct gatcaagaga
     3121 caggatacta gtggaggaag aaaaaatgag caagggcgag gagctgttca ccggggtggt
     3181 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     3241 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     3301 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcgc
     3361 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     3421 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     3481 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     3541 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     3601 caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacatcga
     3661 ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     3721 cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa
     3781 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccgggt aattagctga
     3841 gcttggactc ctgttgatag atccagtaat gacctcagaa ctccatctgg atttgttcag
     3901 aacgctcggt tgccgccggg cgttttttat tggtgagaat ccaagctagc ttggcgagat
     3961 tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata
     4021 tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct
     4081 ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc
     4141 acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat
     4201 ttcgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca
     4261 ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt
     4321 tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct
     4381 atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt
     4441 tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca
     4501 tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc
     4561 atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg
     4621 atgagtggca gggcggggcg taattttttt aaggcagtta ttggtgccct taaacgcctg
     4681 gggtaatgac tctctagctt gaggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     4741 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
     4801 ctaggagctt gcggccgcca ggatatcgac ccaagtaccg ccacctaaca attcgttcaa
     4861 gccgagatcg gcttcccggc cgacgcgtca attcgagcgc gtcaattcga gggccgcggc
     4921 ctaggcggcc agatctgatc aagagacagt ctagctagaa ttaattcgag tcgaccgatc
     4981 ccgtacacaa gtagcgtcct gaacggaacc tttcccgttt tccaggatct gatcttccat
     5041 gtgacctcct aacatggtaa cgttcatgat aacttctgct cttcatcgtg cggccgactg
     5101 ggctaaatct gtgttctctt cggcggcgct gggtgatcct cgccgtactg cccgcttggt
     5161 taacgtcgcc gcccaattgg caaaatattc tggtaaatca ataaccatct catcagaggg
     5221 tagtgaagcc atgcaggaag gcgcttaccg atttatccgc aatcccaacg tttctgccga
     5281 ggcgatcaga aaggctggcg ccatgcaaac agtcaagttg gctcaggagt ttcccgaact
     5341 gctggccatt gaggacacca cctctttgag ttatcgccac caggtcgccg aagagcttgg
     5401 caagctgggc tctattcagg ataaatcccg cggatggtgg gttcactccg ttctcttgct
     5461 cgaggccacc acattccgca ccgtaggatt actgcatcag gagtggtgga tgcgcccgga
     5521 tgaccctgcc gatgcggatg aaaaggagag tggcaaatgg ctggcagccg ccgcaactag
     5581 ccggttacgc atgggcagca tgatgagcaa cgtgattgcg gtctgtgacc gcgaagccga
     5641 tattcatgct tatctgcagg acaaactggc gcataacgag cgcttcgtgg tgcgctccaa
     5701 gcacccacgc aaggacgtag agtctgggtt gtatctgtac gaccatctga agaaccaacc
     5761 ggagttgggt ggctatcaga tcagcattcc gcaaaagggc gtggtggata aacgcggtaa
     5821 acgtaaaaat cgaccagccc gcaaggcgag cttgagcctg cgcagtgggc gcatcacgct
     5881 aaaacagggg aatatcacgc tcaacgcggt gctggccgag gagattaacc cgcccaaggg
     5941 tgagaccccg ttgaaatggt tgttgctgac cagcgaaccg gtcgagtcgc tagcccaagc
     6001 cttgcgcgtc atcgacattt atacccatcg ctggcggatc gaggagttcc ataaggcatg
     6061 gaaaaccgga gcaggagccg agaggcaacg catggaggag ccggataatc tggagcggat
     6121 ggtctcgatc ctctcgtttg ttgcggtcag gctgttacag ctcagagaaa gcttcacgct
     6181 gccgcaagca ctcagggcgc aagggctgct aaaggaagcg gaacacgtag aaagccagtc
     6241 cgcagaaacg gtgctgaccc cggatgaatg tcagctactg ggctatctgg acaagggaaa
     6301 acgcaagcgc aaagagaaag caggtagctt gcagtgggct tacatggcga tagctagact
     6361 gggcggtttt atggacagca agcgaaccgg aattgccagc tggggcgccc tctgggaagg
     6421 ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc tgatggcgca
     6481 ggggatcaag atctgatcgg tcgatcgaat taattcccaa ttcttgaaga cgaaagggcc
     6541 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     6601 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     6661 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     6721 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     6781 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     6841 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     6901 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     6961 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     7021 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     7081 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     7141 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     7201 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     7261 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     7321 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     7381 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     7441 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     7501 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     7561 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     7621 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc cggggaattg
     7681 gggatcaatt ccgttttccg ctgcataacc ctgcttcggg gtcattatag cgattttttc
     7741 ggtatatcca tccttt
//
