LOCUS       sequence-226573-        8474 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 8474)
  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 8474)
  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: pMRE157
FEATURES             Location/Qualifiers
     source          1..8474
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..3,4..6,7..720)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(46..67)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     654..675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      737..831
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             875..1534
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(941..960)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1599..1685
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2024..2818)
                     /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(2135..2154)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2745..2764
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      4298..5069
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             5070..5732
                     /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     6426..6448
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6440..6457
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6441..6457
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6467..6486
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6467..6485
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      6528..6614
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6711..6797
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6894..6980
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7077..7163
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             7319..8113
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     GQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDVGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
       61 ggcgacgtaa acggccacaa gttcagcgtc cgcggcgagg gcgagggcga tgccaccaac
      121 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      181 ctcgtgacca ccttcggcta cggcgtggcc tgcttcagcc gctaccccga ccacatgaag
      241 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatctct
      301 ttcaaggacg acggtaccta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      361 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      421 aagctggagt acaacttcaa cagccactac gtctatatca cggccgacaa gcagaagaac
      481 tgcatcaagg ctaacttcaa gatccgccac aacgttgagg acggcagcgt gcagctcgcc
      541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      601 tacctgagcc atcagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      661 ctgctggagt tcgtgaccgc cgccgggatt acacatggca tggacgagct gtacaagtaa
      721 taattagctg agcttggact cctgttgata gatccagtaa tgacctcaga actccatctg
      781 gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaagctag
      841 cttggcgaga ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac
      901 caccgttgat atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc
      961 tcaatgtacc tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa
     1021 gaaaaataag cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc
     1081 tcatccggaa tttcgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca
     1141 cccttgttac accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata
     1201 ccacgacgat ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga
     1261 aaacctggcc tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc
     1321 ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc
     1381 cgttttcacc atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat
     1441 tcaggttcat catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca
     1501 acagtactgc gatgagtggc agggcggggc gtaatttttt taaggcagtt attggtgccc
     1561 ttaaacgcct ggggtaatga ctctctagct tgaggcatca aataaaacga aaggctcagt
     1621 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     1681 caaatccgcc gctaggagct tgcggccgcc agtgtgatgg atatctgcat aaaaactgtt
     1741 gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga tgaacctgaa
     1801 tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg ggggtgggcg
     1861 aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg
     1921 attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg
     1981 ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa
     2041 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
     2101 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
     2161 cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
     2221 tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt
     2281 cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt
     2341 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc
     2401 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca
     2461 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct
     2521 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca
     2581 cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca
     2641 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg
     2701 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga
     2761 atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc
     2821 gaaacgatcc tcatcctgtc tcttgatcag atcttgatcc cctgcgccat cagatccttg
     2881 gcggcaagaa agccatccag tttactttgc agggcttccc aaccttacca gagggcgccc
     2941 cagctggcaa ttccggttcg cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg
     3001 taagcccact gcaagctacc tgctttctct ttgcgcttgc gttttccctt gtccagatag
     3061 cccagtagct gacattcatc ccaggtggca cttttcgggg aaatgtgcgc gcccgcgttc
     3121 ctgctggcgc tgggcctgtt tctggcgctg gacttcccgc tgttccgtca gcagcttttc
     3181 gcccacggcc ttgatgatcg cggcggcctt ggcctgcata tcccgattca acggccccag
     3241 ggcgtccaga acgggcttca ggcgctcccg aaggtctcgg gccgtctctt gggcttgatc
     3301 ggccttcttg cgcatctcac gcgctcctgc ggcggcctgt agggcaggct catacccctg
     3361 ccgaaccgct tttgtcagcc ggtcggccac ggcttccggc gtctcaacgc gctttgagat
     3421 tcccagcttt tcggccaatc cctgcggtgc ataggcgcgt ggctcgaccg cttgcgggct
     3481 gatggtgacg tggcccactg gtggccgctc cagggcctcg tagaacgcct gaatgcgcgt
     3541 gtgacgtgcc ttgctgccct cgatgccccg ttgcagccct agatcggcca cagcggccgc
     3601 aaacgtggtc tggtcgcggg tcatctgcgc tttgttgccg atgaactcct tggccgacag
     3661 cctgccgtcc tgcgtcagcg gcaccacgaa cgcggtcatg tgcgggctgg tttcgtcacg
     3721 gtggatgctg gccgtcacga tgcgatccgc cccgtacttg tccgccagcc acttgtgcgc
     3781 cttctcgaag aacgccgcct gctgttcttg gctggccgac ttccaccatt ccgggctggc
     3841 cgtcatgacg tactcgaccg ccaacacagc gtccttgcgc cgcttctctg gcagcaactc
     3901 gcgcagtcgg cccatcgctt catcggtgct gctggccgcc cagtgctcgt tctctggcgt
     3961 cctgctggcg tcagcgttgg gcgtctcgcg ctcgcggtag gcgtgcttga gactggccgc
     4021 cacgttgccc attttcgcca gcttcttgca tcgcatgatc gcgtatgccg ccatgcctgc
     4081 ccctcccttt tggtgtccaa ccggctcgac gggggcagcg caaggcggtg cctccggcgg
     4141 gccactcaat gcttgagtat actcactaga ctttgcttcg caaagtcgtg accgcctacg
     4201 gcggctgcgg cgccctacgg gcttgctctc cgggcttcgc cctgcgcggt cgctgcgctc
     4261 ccttgccagc ccgtggatat gtggacgatg gccgcgagcg gccaccggct ggctcgcttc
     4321 gctcggcccg tggacaaccc tgctggacaa gctgatggac aggctgcgcc tgcccacgag
     4381 cttgaccaca gggattgccc accggctacc cagccttcga ccacataccc accggctcca
     4441 actgcgcggc ctgcggcctt gccccatcaa tttttttaat tttctctggg gaaaagcctc
     4501 cggcctgcgg cctgcgcgct tcgcttgccg gttggacacc aagtggaagg cgggtcaagg
     4561 ctcgcgcagc gaccgcgcag cggcttggcc ttgacgcgcc tggaacgacc caagcctatg
     4621 cgagtggggg cagtcgaagg gcgaagcccg cccgcctgcc ccccgagcct cacggcggcg
     4681 agtgcggggg ttccaagggg gcagcgccac cttgggcaag gccgaaggcc gcgcagtcga
     4741 tcaacaagcc ccggaggggc cactttttgc cggaggggga gccgcgccga aggcgtgggg
     4801 gaaccccgca ggggtgccct tctttgggca ccaaagaact agatataggg cgaaatgcga
     4861 aagacttaaa aatcaacaac ttaaaaaagg ggggtacgca acagctcatt gcggcacccc
     4921 ccgcaatagc tcattgcgta ggttaaagaa aatctgtaat tgactgccac ttttacgcaa
     4981 cgcataattg ttgtcgcgct gccgaaaagt tgcagctgat tgcgcatggt gccgcaaccg
     5041 tgcggcaccc ctaccgcatg gagataagca tggccacgca gtccagagaa atcggcattc
     5101 aagccaagaa caagcccggt cactgggtgc aaacggaacg caaagcgcat gaggcgtggg
     5161 ccgggcttat tgcgaggaaa cccacggcgg caatgctgct gcatcacctc gtggcgcaga
     5221 tgggccacca gaacgccgtg gtggtcagcc agaagacact ttccaagctc atcggacgtt
     5281 ctttgcggac ggtccaatac gcagtcaagg acttggtggc cgagcgctgg atctccgtcg
     5341 tgaagctcaa cggccccggc accgtgtcgg cctacgtggt caatgaccgc gtggcgtggg
     5401 gccagccccg cgaccagttg cgcctgtcgg tgttcagtgc cgccgtggtg gttgatcacg
     5461 acgaccagga cgaatcgctg ttggggcatg gcgacctgcg ccgcatcccg accctgtatc
     5521 cgggcgagca gcaactaccg accggccccg gcgaggagcc gcccagccag cccggcattc
     5581 cgggcatgga accagacctg ccagccttga ccgaaacgga ggaatgggaa cggcgcgggc
     5641 agcagcgcct gccgatgccc gatgagccgt gttttctgga cgatggcgag ccgttggagc
     5701 cgccgacacg ggtcacgctg ccgcgccggt agcacttggg ttgcgcagca acccgtaagt
     5761 gcgctgttcc agactatcgg ctgtagccgc ctcgccgccc tataccttgt ctgcctcccc
     5821 gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct gaatggaagc cggcggcacc
     5881 tcgctaacgg attcaccgtt tttatcaggc tctgggaggc agaataaatg atcatatcgt
     5941 caattattac ctccacgggg agagcctgag caaactggcc tcaggcattt gagaagcaca
     6001 cggtcacact gcttccggta gtcaataaac cggtaaacca gcaatagaca taagcggcta
     6061 tttaacgacc ctgccctgaa ccgacgaccg ggtcgaattt gctttcgaat ttctgccatt
     6121 catccgctta ttatcactta ttcaggcgta gcaaccaggc gtttaagggc accaataact
     6181 gccttaaaaa aattacgccc cgccctgcca ctcatcgcag tacggcctat tggttaaaaa
     6241 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     6301 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     6361 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     6421 gttttcccag tcacgacgtt gtaaaacgac ggccagtgag cgcgcgtaat acgactcact
     6481 atagggcgaa ttggagctgg ggccgcaatt cccaattccc caggcatcaa ataaaacgaa
     6541 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     6601 tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt
     6661 ggcgggcagg acgcccgcca taaactgcca ggaattaatt ccccaggcat caaataaaac
     6721 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     6781 tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag
     6841 ggtggcgggc aggacgcccg ccataaactg ccaggaatta attccccagg catcaaataa
     6901 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     6961 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg
     7021 gagggtggcg ggcaggacgc ccgccataaa ctgccaggaa ttaattcccc aggcatcaaa
     7081 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga
     7141 acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc
     7201 ccggagggtg gcgggcagga cgcccgccat aaactgccag gaattgggga tcggattcga
     7261 gctaagaatt cccttggggt atctttaaat actgtagaaa agaggaagga aataataaat
     7321 ggctaaaatg agaatatcac cggaattgaa aaaactgatc gaaaaatacc gctgcgtaaa
     7381 agatacggaa ggaatgtctc ctgctaaggt atataagctg gtgggagaaa atgaaaacct
     7441 atatttaaaa atgacggaca gccggtataa agggaccacc tatgatgtgg aacgggaaaa
     7501 ggacatgatg ctatggctgg aaggaaagct gcctgttcca aaggtcctgc actttgaacg
     7561 gcatgatggc tggagcaatc tgctcatgag tgaggccgat ggcgtccttt gctcggaaga
     7621 gtatgaagat ggacaaagcc ctgaaaagat tatcgagctg tatgcggagt gcatcaggct
     7681 ctttcactcc atcgacatat cggattgtcc ctatacgaat agcttagaca gccgcttagc
     7741 cgaattggat tacttactga ataacgatct ggccgatgtg gattgcgaaa actgggaaga
     7801 agacactcca tttaaagatc cgcgcgagct gtatgatttt ttaaagacgg aaaagcccga
     7861 agaggaactt gtcttttccc acggcgacct gggagacagc aacatctttg tgaaagatgg
     7921 caaagtaagt ggctttattg atcttgggag aagcggcagg gcggacaagt ggtatgacat
     7981 tgccttctgc gtccggtcga tcagggagga tgtcggggaa gaacagtatg tcgagctatt
     8041 ttttgactta ctggggatca agcctgattg ggagaaaata aaatattata ttttactgga
     8101 tgaattgttt tagagctcgc acgctgccgc aagcactcag ggcgcaaggg ctgctaaagg
     8161 aagcggaaca cgtagaaagc cagtccgcag aaacggtgct gaccccggat gaatgtcagc
     8221 tactgggcta tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt agcttgcagt
     8281 gggcttacat ggcgatagct agactgggcg gttttatgga cagcaagcga accggaattg
     8341 ccagctgggg cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc
     8401 ttgccgccaa ggatctgatg gcgcagggga tcaagatctg atcaagagac aggatactag
     8461 tggaggaaga aaaa
//
