LOCUS       sequence-226381-        7600 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 7600)
  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 7600)
  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: pMRE135
FEATURES             Location/Qualifiers
     source          1..7600
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..696
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(134..152)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     402..421
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      717..811
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             855..1514
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(921..940)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1579..1665
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2004..2798)
                     /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(2115..2134)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2725..2744
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      4278..5049
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             5050..5712
                     /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     6406..6428
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6420..6437
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6421..6437
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6447..6466
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6447..6465
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      6508..6594
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6691..6777
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6874..6960
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7057..7143
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 atggtgagca agggcgaggc agtgatcaag gagttcatgc ggttcaaggt gcacatggag
       61 ggctccatga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
      121 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt ctcctgggac
      181 atcctgtccc ctcagttcat gtacggctcc agggccttca tcaagcaccc cgccgacatc
      241 cccgactact ataagcagtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
      301 gaggacggcg gcgccgtgac cgtgacccag gacacctccc tggaggacgg caccctgatc
      361 tacaaggtga agctccgcgg caccaacttc cctcctgacg gccccgtaat gcagaagaag
      421 acaatgggct gggaagcgtc caccgagcgg ttgtaccccg aggacggcgt gctgaagggc
      481 gacattaaga tggccctgcg cctgaaggac ggcggccgct acctggcgga cttcaagacc
      541 acctacaagg ccaagaagcc cgtgcagatg cccggcgcct acaacgtcga ccgcaagttg
      601 gacatcacct cccacaacga ggactacacc gtggtggaac agtacgaacg ctccgagggc
      661 cgccactcca ccggcggcat ggacgagctg tacaagtaag taattagctg agcttggact
      721 cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg
      781 ttgccgccgg gcgtttttta ttggtgagaa tccaagctag cttggcgaga ttttcaggag
      841 ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat
      901 ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga
      961 ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt
     1021 atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa tttcgtatgg
     1081 caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc
     1141 atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt
     1201 ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta
     1261 aagggtttat tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt
     1321 ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat
     1381 attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt
     1441 gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc
     1501 agggcggggc gtaatttttt taaggcagtt attggtgccc ttaaacgcct ggggtaatga
     1561 ctctctagct tgaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     1621 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gctaggagct
     1681 tgcggccgcc agtgtgatgg atatctgcat aaaaactgtt gtaattcatt aagcattctg
     1741 ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc
     1801 ttgtcgcctt gcgtataata tttgcccatg ggggtgggcg aagaactcca gcatgagatc
     1861 cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc
     1921 atagaaggcg gcggtggaat cgaaatctcg tgatggcagg ttgggcgtcg cttggtcggt
     1981 catttcgaac cccagagtcc cgctcagaag aactcgtcaa gaaggcgata gaaggcgatg
     2041 cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc ccattcgccg
     2101 ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg gtccgccaca
     2161 cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat gatattcggc
     2221 aagcaggcat cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg cgccttgagc
     2281 ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc atcctgatcg
     2341 acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc ttggtggtcg
     2401 aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc catgatggat
     2461 actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac ttcgcccaat
     2521 agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca aggaacgccc
     2581 gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca gttcattcag ggcaccggac
     2641 aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa cacggcggca
     2701 tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc cacccaagcg
     2761 gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc tcatcctgtc
     2821 tcttgatcag atcttgatcc cctgcgccat cagatccttg gcggcaagaa agccatccag
     2881 tttactttgc agggcttccc aaccttacca gagggcgccc cagctggcaa ttccggttcg
     2941 cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg taagcccact gcaagctacc
     3001 tgctttctct ttgcgcttgc gttttccctt gtccagatag cccagtagct gacattcatc
     3061 ccaggtggca cttttcgggg aaatgtgcgc gcccgcgttc ctgctggcgc tgggcctgtt
     3121 tctggcgctg gacttcccgc tgttccgtca gcagcttttc gcccacggcc ttgatgatcg
     3181 cggcggcctt ggcctgcata tcccgattca acggccccag ggcgtccaga acgggcttca
     3241 ggcgctcccg aaggtctcgg gccgtctctt gggcttgatc ggccttcttg cgcatctcac
     3301 gcgctcctgc ggcggcctgt agggcaggct catacccctg ccgaaccgct tttgtcagcc
     3361 ggtcggccac ggcttccggc gtctcaacgc gctttgagat tcccagcttt tcggccaatc
     3421 cctgcggtgc ataggcgcgt ggctcgaccg cttgcgggct gatggtgacg tggcccactg
     3481 gtggccgctc cagggcctcg tagaacgcct gaatgcgcgt gtgacgtgcc ttgctgccct
     3541 cgatgccccg ttgcagccct agatcggcca cagcggccgc aaacgtggtc tggtcgcggg
     3601 tcatctgcgc tttgttgccg atgaactcct tggccgacag cctgccgtcc tgcgtcagcg
     3661 gcaccacgaa cgcggtcatg tgcgggctgg tttcgtcacg gtggatgctg gccgtcacga
     3721 tgcgatccgc cccgtacttg tccgccagcc acttgtgcgc cttctcgaag aacgccgcct
     3781 gctgttcttg gctggccgac ttccaccatt ccgggctggc cgtcatgacg tactcgaccg
     3841 ccaacacagc gtccttgcgc cgcttctctg gcagcaactc gcgcagtcgg cccatcgctt
     3901 catcggtgct gctggccgcc cagtgctcgt tctctggcgt cctgctggcg tcagcgttgg
     3961 gcgtctcgcg ctcgcggtag gcgtgcttga gactggccgc cacgttgccc attttcgcca
     4021 gcttcttgca tcgcatgatc gcgtatgccg ccatgcctgc ccctcccttt tggtgtccaa
     4081 ccggctcgac gggggcagcg caaggcggtg cctccggcgg gccactcaat gcttgagtat
     4141 actcactaga ctttgcttcg caaagtcgtg accgcctacg gcggctgcgg cgccctacgg
     4201 gcttgctctc cgggcttcgc cctgcgcggt cgctgcgctc ccttgccagc ccgtggatat
     4261 gtggacgatg gccgcgagcg gccaccggct ggctcgcttc gctcggcccg tggacaaccc
     4321 tgctggacaa gctgatggac aggctgcgcc tgcccacgag cttgaccaca gggattgccc
     4381 accggctacc cagccttcga ccacataccc accggctcca actgcgcggc ctgcggcctt
     4441 gccccatcaa tttttttaat tttctctggg gaaaagcctc cggcctgcgg cctgcgcgct
     4501 tcgcttgccg gttggacacc aagtggaagg cgggtcaagg ctcgcgcagc gaccgcgcag
     4561 cggcttggcc ttgacgcgcc tggaacgacc caagcctatg cgagtggggg cagtcgaagg
     4621 gcgaagcccg cccgcctgcc ccccgagcct cacggcggcg agtgcggggg ttccaagggg
     4681 gcagcgccac cttgggcaag gccgaaggcc gcgcagtcga tcaacaagcc ccggaggggc
     4741 cactttttgc cggaggggga gccgcgccga aggcgtgggg gaaccccgca ggggtgccct
     4801 tctttgggca ccaaagaact agatataggg cgaaatgcga aagacttaaa aatcaacaac
     4861 ttaaaaaagg ggggtacgca acagctcatt gcggcacccc ccgcaatagc tcattgcgta
     4921 ggttaaagaa aatctgtaat tgactgccac ttttacgcaa cgcataattg ttgtcgcgct
     4981 gccgaaaagt tgcagctgat tgcgcatggt gccgcaaccg tgcggcaccc ctaccgcatg
     5041 gagataagca tggccacgca gtccagagaa atcggcattc aagccaagaa caagcccggt
     5101 cactgggtgc aaacggaacg caaagcgcat gaggcgtggg ccgggcttat tgcgaggaaa
     5161 cccacggcgg caatgctgct gcatcacctc gtggcgcaga tgggccacca gaacgccgtg
     5221 gtggtcagcc agaagacact ttccaagctc atcggacgtt ctttgcggac ggtccaatac
     5281 gcagtcaagg acttggtggc cgagcgctgg atctccgtcg tgaagctcaa cggccccggc
     5341 accgtgtcgg cctacgtggt caatgaccgc gtggcgtggg gccagccccg cgaccagttg
     5401 cgcctgtcgg tgttcagtgc cgccgtggtg gttgatcacg acgaccagga cgaatcgctg
     5461 ttggggcatg gcgacctgcg ccgcatcccg accctgtatc cgggcgagca gcaactaccg
     5521 accggccccg gcgaggagcc gcccagccag cccggcattc cgggcatgga accagacctg
     5581 ccagccttga ccgaaacgga ggaatgggaa cggcgcgggc agcagcgcct gccgatgccc
     5641 gatgagccgt gttttctgga cgatggcgag ccgttggagc cgccgacacg ggtcacgctg
     5701 ccgcgccggt agcacttggg ttgcgcagca acccgtaagt gcgctgttcc agactatcgg
     5761 ctgtagccgc ctcgccgccc tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg
     5821 gagccgggcc acctcgacct gaatggaagc cggcggcacc tcgctaacgg attcaccgtt
     5881 tttatcaggc tctgggaggc agaataaatg atcatatcgt caattattac ctccacgggg
     5941 agagcctgag caaactggcc tcaggcattt gagaagcaca cggtcacact gcttccggta
     6001 gtcaataaac cggtaaacca gcaatagaca taagcggcta tttaacgacc ctgccctgaa
     6061 ccgacgaccg ggtcgaattt gctttcgaat ttctgccatt catccgctta ttatcactta
     6121 ttcaggcgta gcaaccaggc gtttaagggc accaataact gccttaaaaa aattacgccc
     6181 cgccctgcca ctcatcgcag tacggcctat tggttaaaaa atgagctgat ttaacaaaaa
     6241 tttaacgcga attttaacaa aatattaacg cttacaattt ccattcgcca ttcaggctgc
     6301 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     6361 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     6421 gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttggagctgg
     6481 ggccgcaatt cccaattccc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     6541 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
     6601 ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca
     6661 taaactgcca ggaattaatt ccccaggcat caaataaaac gaaaggctca gtcgaaagac
     6721 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     6781 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg
     6841 ccataaactg ccaggaatta attccccagg catcaaataa aacgaaaggc tcagtcgaaa
     6901 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     6961 ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc
     7021 ccgccataaa ctgccaggaa ttaattcccc aggcatcaaa taaaacgaaa ggctcagtcg
     7081 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
     7141 aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga
     7201 cgcccgccat aaactgccag gaattgggga tcggattcga gctcgcacgc tgccgcaagc
     7261 actcagggcg caagggctgc taaaggaagc ggaacacgta gaaagccagt ccgcagaaac
     7321 ggtgctgacc ccggatgaat gtcagctact gggctatctg gacaagggaa aacgcaagcg
     7381 caaagagaaa gcaggtagct tgcagtgggc ttacatggcg atagctagac tgggcggtct
     7441 tatggacagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag gttgggaagc
     7501 cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc aggggatcaa
     7561 gatctgatca agagacagga tactagtgga ggaagaaaaa
//
