LOCUS       sequence-366486-        7793 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 7793)
  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 7793)
  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-Tn5-163
FEATURES             Location/Qualifiers
     source          1..7793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            152..261
                     /note="incP origin of transfer"
     primer_bind     815..831
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     815..831
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             complement(894..2093)
                     /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(3226..3228,3229..3231,3232..3942)
                     /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(3271..3292)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3532..3551)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3879..3900
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      3963..4057
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             4101..4760
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4167..4186)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      4825..4911
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(6655..6673)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6741..6845
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6846..6914,6915..7706)
                     /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(7064..7083)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 attggggatc aattccgttt tccgctgcat aaccctgctt cggggtcatt atagcgattt
       61 tttcggtata tccatccttt ttcgcacgat atacaggatt ttgccaaagg gttcgtgtag
      121 actttccttg gtgtatccaa cggcgtcagc cgggcaggat aggtgaagta ggcccacccg
      181 cgagcgggtg ttccttcttc actgtccctt attcgcacct ggcggtgctc aacgggaatc
      241 ctgctctgcg aggctggccg gctaccgccg gcgtaacaga tgagggcaag cggatggctg
      301 atgaaaccaa gccaaccagg aagggcagcc cacctatcac ggaattgatc cggccacgat
      361 gcgtccggcg tagaggatct gaagatcagc agttcaacct gttgatagta cgtactaagc
      421 tctcatgttt cacgtactaa gctctcatgt ttaacgtact aagctctcat gtttaacgaa
      481 ctaaaccctc atggctaacg tactaagctc tcatgtttca cgtactaagc tctcatgttt
      541 gaacaataaa attaatataa atcagcaact taaatagcct ctaaggtttt aagttttata
      601 agaaaaaaaa gaatatataa ggcttttaaa gcttttaagg tttaacggtt gtggacaaca
      661 agccagggat gtaacgcact gagaagccct tagagcctct caaagcaatt ttgagtgaca
      721 caggaacact taacggctga catgggaatt aattctgact cttatacaca agtgcggccc
      781 tcgaattgac gcgctccacc gcggtggcgg ccgctctaga actagtggat cccccgggct
      841 gcaggaattc gatatccata aactgccagg aattggggat cggattcgag ctctcagcga
      901 tcggctcgtt gccctgcgcc gctccaaagc ccgcgacgca gcgccggcag gcagagcaag
      961 tagagggcag cgcctgcaat ccatgcccac ccgttccacg ttgttataga agccgcatag
     1021 atcgccgtga agaggagggg tccgacgatc gaggtcaggc tggtgagcgc cgccagtgag
     1081 ccttgcagct gcccctgacg ttcctcatcc acctgcctgg acaacattgc ttgcagcgcc
     1141 ggcattccga tgccacccga agcaagcagg accatgatcg ggaacgccat ccatccccgt
     1201 gtcgcgaagg caagcaggat gtagcctgtg ccgtcggcaa tcattccgag catgagtgcc
     1261 cgcctttcgc cgagccgggc ggctacaggg ccggtgatca ttgcctgggc gagtgaatgc
     1321 agaatgccaa atgcggcaag cgaaatgccg atcgtggtcg cgtcccagtg aaagcgatcc
     1381 tcgccgaaaa tgacccaaag cgcggccggc acctgtccga caagttgcat gatgaagaag
     1441 accgccatca gggcggcgac gacggtcatg ccccgggccc accggaacga agcgagcggg
     1501 ttgagagcct cccggcgtaa cggccggcgt tcgcctttgt gcgactccgg caaaaggaaa
     1561 cagcccgtca ggaaattgag gccgttcaag gctgccgcgg cgaagaacgg agcgtggggg
     1621 gagaaaccgc ccatcagccc accgagcaca ggtcccgcga ccatcccgaa cccgaaacag
     1681 gcgctcatga agccgaagtg ccgcgcgcgc tcatcgccat cagtgatatc ggcaatataa
     1741 gcgccggcta ccgccccagt cgccccggtg atgccggcca cgatccgccc gatatagaga
     1801 acccaaagga aaggcgctgt cgccatgatg gcgtagtcga cagtggcgcc ggccagcgag
     1861 acgagcaaga ttggccgccg cccgaaacga tccgacagcg cgcccagcac aggtgcgcag
     1921 gcaaattgca ccaacgcata cagcgccagc agaatgccat agtgggcggt gacgtcgttc
     1981 gagtgaacca gatcgcgcag gaggcccggc agcaccggca taatcaggcc gatgccgaca
     2041 gcgtcgagcg cgacagtgct cagaattacg atcaggggta tgttgggttt cacgtctggc
     2101 ctccggacca gcggaggctg gtccgattga acgcgcggat tctttatcac tgataagttg
     2161 gtggacatat tatgtttatc agtgataaag tgtcaagcat gacaaagttg cagccgaata
     2221 cagtgatccg tgccgccctg gacctgttga acgaggtcgg cgtagacggt ctgacgacac
     2281 gcaaactggc ggaacggttg ggggttcagc agccggcgct ttactggcac ttcaggaaca
     2341 agcgggcgct gctcgacgca ctggccgaag ccatgctggc ggagaatcat acgcattcgg
     2401 tgccgagagc cgacgacgac tggcgctcat ttctgatcgg gaatgcccgc agcttcaggc
     2461 aggcgctgct cgcctaccgc gatggcgcgc gcatccatgc cggcacgcga ccgggcgcac
     2521 cgcagatgga aacggccgac gcgcagcttc gcttcctctg cgaggcgggt ttttcggccg
     2581 gggacgccgt caatgcgctg atgacaatca gctacttcac tgttggggcc gtgcttgagg
     2641 agcaggccgg cgacagcgat gccggcgagc gcggcggcac cgttgaacag gctccgctct
     2701 cgccgctgtt gcgggccgcg atagacgcct tcgacgaagc cggtccggac gcagcgttcg
     2761 agcagggact cgcggtgatt gtcgatggat tggcgaaaag gaggctcgtt gtcaggaacg
     2821 ttgaaggacc gagaaagggt gctaagaaac cattattatc atgaagctcg cacgctgccg
     2881 caagcactca gggcgcaagg gctgctaaag gaagcggaac acgtagaaag ccagtccgca
     2941 gaaacggtgc tgaccccgga tgaatgtcag ctactgggct atctggacaa gggaaaacgc
     3001 aagcgcaaag agaaagcagg tagcttgcag tgggcttaca tggcgatagc tagactgggc
     3061 ggttttatgg acagcaagcg aaccggaatt gccagctggg gcgccctctg gtaaggttgg
     3121 gaagccctgc aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg
     3181 atcaagatct gatcaagaga caggatacta gtggaggaag aaaaaatggt gagcaagggc
     3241 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     3301 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     3361 aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     3421 ggctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     3481 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     3541 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     3601 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     3661 tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     3721 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag
     3781 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     3841 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     3901 accgccgccg ggatcactct cggcatggac gagctgtaca agtaagtaat tagctgagct
     3961 tggactcctg ttgatagatc cagtaatgac ctcagaactc catctggatt tgttcagaac
     4021 gctcggttgc cgccgggcgt tttttattgg tgagaatcca agctagcttg gcgagatttt
     4081 caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat
     4141 cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata
     4201 accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca
     4261 agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaatttc
     4321 gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg
     4381 ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc
     4441 ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt
     4501 tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca
     4561 ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg
     4621 gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg
     4681 ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg
     4741 agtggcaggg cggggcgtaa tttttttaag gcagttattg gtgcccttaa acgcctgggg
     4801 taatgactct ctagcttgag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
     4861 tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccgcta
     4921 ggagcttgcg gccgccagga tatcgaccca agtaccgcca cctaacaatt cgttcaagcc
     4981 gagatcggct tcccggccga cgcgtcaatt cgagcgcgtc aattcgaggg ccgcggccta
     5041 ggcggccaga tctgatcaag agacagtcta gctagaatta attcgagtcg accgatcccg
     5101 tacacaagta gcgtcctgaa cggaaccttt cccgttttcc aggatctgat cttccatgtg
     5161 acctcctaac atggtaacgt tcatgataac ttctgctctt catcgtgcgg ccgactgggc
     5221 taaatctgtg ttctcttcgg cggcgctggg tgatcctcgc cgtactgccc gcttggttaa
     5281 cgtcgccgcc caattggcaa aatattctgg taaatcaata accatctcat cagagggtag
     5341 tgaagccatg caggaaggcg cttaccgatt tatccgcaat cccaacgttt ctgccgaggc
     5401 gatcagaaag gctggcgcca tgcaaacagt caagttggct caggagtttc ccgaactgct
     5461 ggccattgag gacaccacct ctttgagtta tcgccaccag gtcgccgaag agcttggcaa
     5521 gctgggctct attcaggata aatcccgcgg atggtgggtt cactccgttc tcttgctcga
     5581 ggccaccaca ttccgcaccg taggattact gcatcaggag tggtggatgc gcccggatga
     5641 ccctgccgat gcggatgaaa aggagagtgg caaatggctg gcagccgccg caactagccg
     5701 gttacgcatg ggcagcatga tgagcaacgt gattgcggtc tgtgaccgcg aagccgatat
     5761 tcatgcttat ctgcaggaca aactggcgca taacgagcgc ttcgtggtgc gctccaagca
     5821 cccacgcaag gacgtagagt ctgggttgta tctgtacgac catctgaaga accaaccgga
     5881 gttgggtggc tatcagatca gcattccgca aaagggcgtg gtggataaac gcggtaaacg
     5941 taaaaatcga ccagcccgca aggcgagctt gagcctgcgc agtgggcgca tcacgctaaa
     6001 acaggggaat atcacgctca acgcggtgct ggccgaggag attaacccgc ccaagggtga
     6061 gaccccgttg aaatggttgt tgctgaccag cgaaccggtc gagtcgctag cccaagcctt
     6121 gcgcgtcatc gacatttata cccatcgctg gcggatcgag gagttccata aggcatggaa
     6181 aaccggagca ggagccgaga ggcaacgcat ggaggagccg gataatctgg agcggatggt
     6241 ctcgatcctc tcgtttgttg cggtcaggct gttacagctc agagaaagct tcacgctgcc
     6301 gcaagcactc agggcgcaag ggctgctaaa ggaagcggaa cacgtagaaa gccagtccgc
     6361 agaaacggtg ctgaccccgg atgaatgtca gctactgggc tatctggaca agggaaaacg
     6421 caagcgcaaa gagaaagcag gtagcttgca gtgggcttac atggcgatag ctagactggg
     6481 cggttttatg gacagcaagc gaaccggaat tgccagctgg ggcgccctct gggaaggttg
     6541 ggaagccctg caaagtaaac tggatggctt tcttgccgcc aaggatctga tggcgcaggg
     6601 gatcaagatc tgatcggtcg atcgaattaa ttcccaattc ttgaagacga aagggcctcg
     6661 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     6721 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     6781 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     6841 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     6901 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     6961 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     7021 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     7081 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     7141 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     7201 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     7261 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     7321 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     7381 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     7441 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     7501 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     7561 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     7621 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     7681 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     7741 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatccgg gga
//
