LOCUS       sequence-227232-        7783 bp    DNA     circular SYN 14-APR-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 Apr 14, 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-Tn5-160
FEATURES             Location/Qualifiers
     source          1..7783
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             484..1194
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     terminator      1211..1305
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1349..2008
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1415..1434)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2073..2159
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3903..3921)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3989..4093
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4094..4162,4163..4954)
                     /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(4312..4331)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     oriT            5193..5302
                     /note="incP origin of transfer"
     primer_bind     5856..5872
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5856..5872
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             complement(5935..7134)
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
ORIGIN
        1 gtccggacgc agcgttcgag cagggactcg cggtgattgt cgatggattg gcgaaaagga
       61 ggctcgttgt caggaacgtt gaaggaccga gaaagggtgc taagaaacca ttattatcat
      121 gaagctcgca cgctgccgca agcactcagg gcgcaagggc tgctaaagga agcggaacac
      181 gtagaaagcc agtccgcaga aacggtgctg accccggatg aatgtcagct actgggctat
      241 ctggacaagg gaaaacgcaa gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg
      301 gcgatagcta gactgggcgg ttttatggac agcaagcgaa ccggaattgc cagctggggc
      361 gccctctggt aaggttggga agccctgcaa agtaaactgg atggctttct tgccgccaag
      421 gatctgatgg cgcaggggat caagatctga tcaagagaca ggatactagt ggaggaagaa
      481 aaaatggtgt ctaagggcga agagctgatt aaggagaaca tgcacatgaa gctgtacatg
      541 gagggcaccg tggacaacca tcacttcaag tgcacatccg agggcgaagg caagccctac
      601 gagggcaccc agaccatgag aatcaaggtg gtcgagggcg gccctctccc cttcgccttc
      661 gacatcctgg ctactagctt cctctacggc agcaagacct tcatcaacca cacccagggc
      721 atccccgact tcttcaagca gtccttccct gagggcttca catgggagag agtcaccaca
      781 tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc
      841 atctacaacg tcaagatcag aggggtgaac ttcacatcca acggccctgt gatgcagaag
      901 aaaacactcg gctgggaggc cttcaccgag acgctgtacc ccgctgacgg cggcctggaa
      961 ggcagaaacg acatggccct gaagctcgtg ggcgggagcc atctgatcgc aaacgccaag
     1021 accacatata gatccaagaa acccgctaag aacctcaaga tgcctggcgt ctactatgtg
     1081 gactacagac tggaaagaat caaggaggcc aacaacgaga cctacgtcga gcagcacgag
     1141 gtggcagtgg ccagatactg cgacctccct agcaaactgg ggcacaagct taattaatta
     1201 gctgagcttg gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg
     1261 ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag ctagcttggc
     1321 gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt
     1381 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg
     1441 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa
     1501 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc
     1561 ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg
     1621 ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga
     1681 cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct
     1741 ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt
     1801 gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt
     1861 caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt
     1921 tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta
     1981 ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattggt gcccttaaac
     2041 gcctggggta atgactctct agcttgaggc atcaaataaa acgaaaggct cagtcgaaag
     2101 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
     2161 cgccgctagg agcttgcggc cgccaggata tcgacccaag taccgccacc taacaattcg
     2221 ttcaagccga gatcggcttc ccggccgacg cgtcaattcg agcgcgtcaa ttcgagggcc
     2281 gcggcctagg cggccagatc tgatcaagag acagtctagc tagaattaat tcgagtcgac
     2341 cgatcccgta cacaagtagc gtcctgaacg gaacctttcc cgttttccag gatctgatct
     2401 tccatgtgac ctcctaacat ggtaacgttc atgataactt ctgctcttca tcgtgcggcc
     2461 gactgggcta aatctgtgtt ctcttcggcg gcgctgggtg atcctcgccg tactgcccgc
     2521 ttggttaacg tcgccgccca attggcaaaa tattctggta aatcaataac catctcatca
     2581 gagggtagtg aagccatgca ggaaggcgct taccgattta tccgcaatcc caacgtttct
     2641 gccgaggcga tcagaaaggc tggcgccatg caaacagtca agttggctca ggagtttccc
     2701 gaactgctgg ccattgagga caccacctct ttgagttatc gccaccaggt cgccgaagag
     2761 cttggcaagc tgggctctat tcaggataaa tcccgcggat ggtgggttca ctccgttctc
     2821 ttgctcgagg ccaccacatt ccgcaccgta ggattactgc atcaggagtg gtggatgcgc
     2881 ccggatgacc ctgccgatgc ggatgaaaag gagagtggca aatggctggc agccgccgca
     2941 actagccggt tacgcatggg cagcatgatg agcaacgtga ttgcggtctg tgaccgcgaa
     3001 gccgatattc atgcttatct gcaggacaaa ctggcgcata acgagcgctt cgtggtgcgc
     3061 tccaagcacc cacgcaagga cgtagagtct gggttgtatc tgtacgacca tctgaagaac
     3121 caaccggagt tgggtggcta tcagatcagc attccgcaaa agggcgtggt ggataaacgc
     3181 ggtaaacgta aaaatcgacc agcccgcaag gcgagcttga gcctgcgcag tgggcgcatc
     3241 acgctaaaac aggggaatat cacgctcaac gcggtgctgg ccgaggagat taacccgccc
     3301 aagggtgaga ccccgttgaa atggttgttg ctgaccagcg aaccggtcga gtcgctagcc
     3361 caagccttgc gcgtcatcga catttatacc catcgctggc ggatcgagga gttccataag
     3421 gcatggaaaa ccggagcagg agccgagagg caacgcatgg aggagccgga taatctggag
     3481 cggatggtct cgatcctctc gtttgttgcg gtcaggctgt tacagctcag agaaagcttc
     3541 acgctgccgc aagcactcag ggcgcaaggg ctgctaaagg aagcggaaca cgtagaaagc
     3601 cagtccgcag aaacggtgct gaccccggat gaatgtcagc tactgggcta tctggacaag
     3661 ggaaaacgca agcgcaaaga gaaagcaggt agcttgcagt gggcttacat ggcgatagct
     3721 agactgggcg gttttatgga cagcaagcga accggaattg ccagctgggg cgccctctgg
     3781 gaaggttggg aagccctgca aagtaaactg gatggctttc ttgccgccaa ggatctgatg
     3841 gcgcagggga tcaagatctg atcggtcgat cgaattaatt cccaattctt gaagacgaaa
     3901 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     3961 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     4021 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4081 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4141 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4201 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4261 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4321 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4381 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4441 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4501 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4561 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4621 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4681 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4741 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4801 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4861 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4921 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4981 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatccgggg
     5041 aattggggat caattccgtt ttccgctgca taaccctgct tcggggtcat tatagcgatt
     5101 ttttcggtat atccatcctt tttcgcacga tatacaggat tttgccaaag ggttcgtgta
     5161 gactttcctt ggtgtatcca acggcgtcag ccgggcagga taggtgaagt aggcccaccc
     5221 gcgagcgggt gttccttctt cactgtccct tattcgcacc tggcggtgct caacgggaat
     5281 cctgctctgc gaggctggcc ggctaccgcc ggcgtaacag atgagggcaa gcggatggct
     5341 gatgaaacca agccaaccag gaagggcagc ccacctatca cggaattgat ccggccacga
     5401 tgcgtccggc gtagaggatc tgaagatcag cagttcaacc tgttgatagt acgtactaag
     5461 ctctcatgtt tcacgtacta agctctcatg tttaacgtac taagctctca tgtttaacga
     5521 actaaaccct catggctaac gtactaagct ctcatgtttc acgtactaag ctctcatgtt
     5581 tgaacaataa aattaatata aatcagcaac ttaaatagcc tctaaggttt taagttttat
     5641 aagaaaaaaa agaatatata aggcttttaa agcttttaag gtttaacggt tgtggacaac
     5701 aagccaggga tgtaacgcac tgagaagccc ttagagcctc tcaaagcaat tttgagtgac
     5761 acaggaacac ttaacggctg acatgggaat taattctgac tcttatacac aagtgcggcc
     5821 ctcgaattga cgcgctccac cgcggtggcg gccgctctag aactagtgga tcccccgggc
     5881 tgcaggaatt cgatatccat aaactgccag gaattgggga tcggattcga gctctcagcg
     5941 atcggctcgt tgccctgcgc cgctccaaag cccgcgacgc agcgccggca ggcagagcaa
     6001 gtagagggca gcgcctgcaa tccatgccca cccgttccac gttgttatag aagccgcata
     6061 gatcgccgtg aagaggaggg gtccgacgat cgaggtcagg ctggtgagcg ccgccagtga
     6121 gccttgcagc tgcccctgac gttcctcatc cacctgcctg gacaacattg cttgcagcgc
     6181 cggcattccg atgccacccg aagcaagcag gaccatgatc gggaacgcca tccatccccg
     6241 tgtcgcgaag gcaagcagga tgtagcctgt gccgtcggca atcattccga gcatgagtgc
     6301 ccgcctttcg ccgagccggg cggctacagg gccggtgatc attgcctggg cgagtgaatg
     6361 cagaatgcca aatgcggcaa gcgaaatgcc gatcgtggtc gcgtcccagt gaaagcgatc
     6421 ctcgccgaaa atgacccaaa gcgcggccgg cacctgtccg acaagttgca tgatgaagaa
     6481 gaccgccatc agggcggcga cgacggtcat gccccgggcc caccggaacg aagcgagcgg
     6541 gttgagagcc tcccggcgta acggccggcg ttcgcctttg tgcgactccg gcaaaaggaa
     6601 acagcccgtc aggaaattga ggccgttcaa ggctgccgcg gcgaagaacg gagcgtgggg
     6661 ggagaaaccg cccatcagcc caccgagcac aggtcccgcg accatcccga acccgaaaca
     6721 ggcgctcatg aagccgaagt gccgcgcgcg ctcatcgcca tcagtgatat cggcaatata
     6781 agcgccggct accgccccag tcgccccggt gatgccggcc acgatccgcc cgatatagag
     6841 aacccaaagg aaaggcgctg tcgccatgat ggcgtagtcg acagtggcgc cggccagcga
     6901 gacgagcaag attggccgcc gcccgaaacg atccgacagc gcgcccagca caggtgcgca
     6961 ggcaaattgc accaacgcat acagcgccag cagaatgcca tagtgggcgg tgacgtcgtt
     7021 cgagtgaacc agatcgcgca ggaggcccgg cagcaccggc ataatcaggc cgatgccgac
     7081 agcgtcgagc gcgacagtgc tcagaattac gatcaggggt atgttgggtt tcacgtctgg
     7141 cctccggacc agcctccgct ggtccgattg aacgcgcgga ttctttatca ctgataagtt
     7201 ggtggacata ttatgtttat cagtgataaa gtktcaagca tgacaaagtt gcagccgaat
     7261 acagtgatcc gtgccgccct ggacctgttg aacgaggtcg gcgtagacgg tctgacgaca
     7321 cgcaaactgg cggaacggtt gggggttcag cagccggcgc tttactggca cttcaggaac
     7381 aagcgggcgc tgctcgacgc actggccgaa gccatgctgg cggagaatca tacgcattcg
     7441 gtgccgagag ccgacgacga ctggcgctca tttctgatcg ggaatgcccg cagcttcagg
     7501 caggcgctgc tcgcctaccg cgatggcgcg cgcatccatg ccggcacgcg accgggcgca
     7561 ccgcagatgg aaacggccga cgcgcagctt cgcttcctct gcgaggcggg tttttcggcc
     7621 ggggacgccg tcaatgcgct gatgacaatc agctacttca ctgttggggc cgtgcttgag
     7681 gagcaggccg gcgacagcga tgccggcgag cgcggcggca ccgttgaaca ggctccgctc
     7741 tcgccgctgt tgcgggccgc gatagacgcc ttcgacgaag ccg
//
