LOCUS       sequence-367818-        7611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  broad-host range plasmid to constitutively express fluorescent 
            protein genes in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7611)
  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 7611)
  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: pMRE130
FEATURES             Location/Qualifiers
     source          1..7611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1023..1794
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1795..2457
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     primer_bind     3151..3173
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3165..3182
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3166..3182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3192..3211
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3192..3210
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      3253..3339
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3436..3522
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3619..3705
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3802..3888
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             4346..5056
                     /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      5073..5167
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             5211..5870
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5277..5296)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      5935..6021
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(6360..7154)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6471..6490)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7081..7100
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 ctcgggccgt ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct cctgcggcgg
       61 cctgtagggc aggctcatac ccctgccgaa ccgcttttgt cagccggtcg gccacggctt
      121 ccggcgtctc aacgcgcttt gagattccca gcttttcggc caatccctgc ggtgcatagg
      181 cgcgtggctc gaccgcttgc gggctgatgg tgacgtggcc cactggtggc cgctccaggg
      241 cctcgtagaa cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg ccccgttgca
      301 gccctagatc ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc tgcgctttgt
      361 tgccgatgaa ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc acgaacgcgg
      421 tcatgtgcgg gctggtttcg tcacggtgga tgctggccgt cacgatgcga tccgccccgt
      481 acttgtccgc cagccacttg tgcgccttct cgaagaacgc cgcctgctgt tcttggctgg
      541 ccgacttcca ccattccggg ctggccgtca tgacgtactc gaccgccaac acagcgtcct
      601 tgcgccgctt ctctggcagc aactcgcgca gtcggcccat cgcttcatcg gtgctgctgg
      661 ccgcccagtg ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc tcgcgctcgc
      721 ggtaggcgtg cttgagactg gccgccacgt tgcccatttt cgccagcttc ttgcatcgca
      781 tgatcgcgta tgccgccatg cctgcccctc ccttttggtg tccaaccggc tcgacggggg
      841 cagcgcaagg cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg
      901 cttcgcaaag tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc
      961 ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc
     1021 gagcggccac cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga
     1081 tggacaggct gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc
     1141 ttcgaccaca tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt
     1201 ttaattttct ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg
     1261 acaccaagtg gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac
     1321 gcgcctggaa cgacccaagc ctatgcgagt gggggcagtc gaagggcgaa gcccgcccgc
     1381 ctgccccccg agcctcacgg cggcgagtgc gggggttcca agggggcagc gccaccttgg
     1441 gcaaggccga aggccgcgca gtcgatcaac aagccccgga ggggccactt tttgccggag
     1501 ggggagccgc gccgaaggcg tgggggaacc ccgcaggggt gcccttcttt gggcaccaaa
     1561 gaactagata tagggcgaaa tgcgaaagac ttaaaaatca acaacttaaa aaaggggggt
     1621 acgcaacagc tcattgcggc accccccgca atagctcatt gcgtaggtta aagaaaatct
     1681 gtaattgact gccactttta cgcaacgcat aattgttgtc gcgctgccga aaagttgcag
     1741 ctgattgcgc atggtgccgc aaccgtgcgg cacccctacc gcatggagat aagcatggcc
     1801 acgcagtcca gagaaatcgg cattcaagcc aagaacaagc ccggtcactg ggtgcaaacg
     1861 gaacgcaaag cgcatgaggc gtgggccggg cttattgcga ggaaacccac ggcggcaatg
     1921 ctgctgcatc acctcgtggc gcagatgggc caccagaacg ccgtggtggt cagccagaag
     1981 acactttcca agctcatcgg acgttctttg cggacggtcc aatacgcagt caaggacttg
     2041 gtggccgagc gctggatctc cgtcgtgaag ctcaacggcc ccggcaccgt gtcggcctac
     2101 gtggtcaatg accgcgtggc gtggggccag ccccgcgacc agttgcgcct gtcggtgttc
     2161 agtgccgccg tggtggttga tcacgacgac caggacgaat cgctgttggg gcatggcgac
     2221 ctgcgccgca tcccgaccct gtatccgggc gagcagcaac taccgaccgg ccccggcgag
     2281 gagccgccca gccagcccgg cattccgggc atggaaccag acctgccagc cttgaccgaa
     2341 acggaggaat gggaacggcg cgggcagcag cgcctgccga tgcccgatga gccgtgtttt
     2401 ctggacgatg gcgagccgtt ggagccgccg acacgggtca cgctgccgcg ccggtagcac
     2461 ttgggttgcg cagcaacccg taagtgcgct gttccagact atcggctgta gccgcctcgc
     2521 cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc
     2581 gacctgaatg gaagccggcg gcacctcgct aacggattca ccgtttttat caggctctgg
     2641 gaggcagaat aaatgatcat atcgtcaatt attacctcca cggggagagc ctgagcaaac
     2701 tggcctcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta
     2761 aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac gaccgggtcg
     2821 aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag gcgtagcaac
     2881 caggcgttta agggcaccaa taactgcctt aaaaaaatta cgccccgccc tgccactcat
     2941 cgcagtacgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     3001 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
     3061 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     3121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     3181 gtgagcgcgc gtaatacgac tcactatagg gcgaattgga gctggggccg caattcccaa
     3241 ttccccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     3301 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
     3361 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggaat
     3421 taattcccca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     3481 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     3541 aacgttgcga agcaacggcc cggagggkgg sgggcaggac gcccgccata aactgccagg
     3601 aattaattcc ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     3661 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
     3721 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
     3781 aggaattaat tccccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     3841 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg
     3901 gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact
     3961 gccaggaatt ggggatcgga ttcgagctcg cacgctgccg caagcactca gggcgcaagg
     4021 gctgctaaag gaagcggaac acgtagaaag ccagtccgca gaaacggtgc tgaccccgga
     4081 tgaatgtcag ctactgggct atctggacaa gggaaaacgc aagcgcaaag agaaagcagg
     4141 tagcttgcag tgggcttaca tggcgatagc tagactgggc ggttttatgg acagcaagcg
     4201 aaccggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc aaagtaaact
     4261 ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagatct gatcaagaga
     4321 caggatacta gtggaggaag aaaaaatggt gtctaagggc gaagagctga ttaaggagaa
     4381 catgcacatg aagctgtaca tggagggcac cgtggacaac catcacttca agtgcacatc
     4441 cgagggcgaa ggcaagccct acgagggcac ccagaccatg agaatcaagg tggtcgaggg
     4501 cggccctctc cccttcgcct tcgacatcct ggctactagc ttcctctacg gcagcaagac
     4561 cttcatcaac cacacccagg gcatccccga cttcttcaag cagtccttcc ctgagggctt
     4621 cacatgggag agagtcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac
     4681 cagcctccag gacggctgcc tcatctacaa cgtcaagatc agaggggtga acttcacatc
     4741 caacggccct gtgatgcaga agaaaacact cggctgggag gccttcaccg agacgctgta
     4801 ccccgctgac ggcggcctgg aaggcagaaa cgacatggcc ctgaagctcg tgggcgggag
     4861 ccatctgatc gcaaacgcca agaccacata tagatccaag aaacccgcta agaacctcaa
     4921 gatgcctggc gtctactatg tggactacag actggaaaga atcaaggagg ccaacaacga
     4981 gacctacgtc gagcagcacg aggtggcagt ggccagatac tgcgacctcc ctagcaaact
     5041 ggggcacaag cttaattaat tagctgagct tggactcctg ttgatagatc cagtaatgac
     5101 ctcagaactc catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg
     5161 tgagaatcca agctagcttg gcgagatttt caggagctaa ggaagctaaa atggagaaaa
     5221 aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg
     5281 catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct
     5341 ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg
     5401 cccgcctgat gaatgctcat ccggaatttc gtatggcaat gaaagacggt gagctggtga
     5461 tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat
     5521 cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg
     5581 tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt
     5641 tcgtctcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg
     5701 acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc
     5761 tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa
     5821 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa tttttttaag
     5881 gcagttattg gtgcccttaa acgcctgggg taatgactct ctagcttgag gcatcaaata
     5941 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     6001 gctctcctga gtaggacaaa tccgccgcta ggagcttgcg gccgccagtg tgatggatat
     6061 ctgcataaaa actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg
     6121 gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg
     6181 cccatggggg tgggcgaaga actccagcat gagatccccg cgctggagga tcatccagcc
     6241 ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa
     6301 atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca gagtcccgct
     6361 cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata
     6421 ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg
     6481 gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat
     6541 ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg
     6601 acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg
     6661 agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta
     6721 cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc
     6781 gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga
     6841 gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca
     6901 gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc
     6961 gctgcctcgt cctgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc
     7021 gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt
     7081 gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca
     7141 tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcagatct tgatcccctg
     7201 cgccatcaga tccttggcgg caagaaagcc atccagttta ctttgcaggg cttcccaacc
     7261 ttaccagagg gcgccccagc tggcaattcc ggttcgcttg ctgtccataa aaccgcccag
     7321 tctagctatc gccatgtaag cccactgcaa gctacctgct ttctctttgc gcttgcgttt
     7381 tcccttgtcc agatagccca gtagctgaca ttcatcccag gtggcacttt tcggggaaat
     7441 gtgcgcgccc gcgttcctgc tggcgctggg cctgtttctg gcgctggact tcccgctgtt
     7501 ccgtcagcag cttttcgccc acggccttga tgatcgcggc ggccttggcc tgcatatccc
     7561 gattcaacgg ccccagggcg tccagaacgg gcttcaggcg ctcccgaagg t
//
