LOCUS       sequence-226579-        8469 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 8469)
  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 8469)
  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: pMRE151
FEATURES             Location/Qualifiers
     source          1..8469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..3,4..717)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(43..64)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(304..323)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     651..672
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      731..825
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             869..1528
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(935..954)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1593..1679
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2018..2812)
                     /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(2129..2148)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2739..2758
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      4292..5063
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             5064..5726
                     /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     6420..6442
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6434..6451
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6435..6451
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6461..6480
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6461..6479
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      6522..6608
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6705..6791
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6888..6974
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7071..7157
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             7313..8107
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     GQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDVGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 atgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
       61 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      121 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      181 gtgaccaccc tgtcctgggg cgtgcagtgc ttcgcccgct accccgacca catgaagcag
      241 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
      301 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
      361 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
      421 ctggagtaca actactttag cgacaacgtc tatatcaccg ccgacaagca gaagaacggc
      481 atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac
      541 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
      601 ctgagcaccc agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg
      661 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaatta
      721 gctgagcttg gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg
      781 ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag ctagcttggc
      841 gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat ataccaccgt
      901 tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg
      961 tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg taaagaaaaa
     1021 taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga atgctcatcc
     1081 ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg ttcacccttg
     1141 ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg aataccacga
     1201 cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg gtgaaaacct
     1261 ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca atccctgggt
     1321 gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg cccccgtttt
     1381 caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg cgattcaggt
     1441 tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat tacaacagta
     1501 ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattggt gcccttaaac
     1561 gcctggggta atgactctct agcttgaggc atcaaataaa acgaaaggct cagtcgaaag
     1621 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
     1681 cgccgctagg agcttgcggc cgccagtgtg atggatatct gcataaaaac tgttgtaatt
     1741 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca
     1801 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catgggggtg ggcgaagaac
     1861 tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg
     1921 aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc
     1981 gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc
     2041 gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt
     2101 cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat
     2161 agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca
     2221 ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca
     2281 tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca
     2341 gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt
     2401 tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat
     2461 cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg
     2521 gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg
     2581 cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc tgcagttcat
     2641 tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc
     2701 ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc
     2761 tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg
     2821 atcctcatcc tgtctcttga tcagatcttg atcccctgcg ccatcagatc cttggcggca
     2881 agaaagccat ccagtttact ttgcagggct tcccaacctt accagagggc gccccagctg
     2941 gcaattccgg ttcgcttgct gtccataaaa ccgcccagtc tagctatcgc catgtaagcc
     3001 cactgcaagc tacctgcttt ctctttgcgc ttgcgttttc ccttgtccag atagcccagt
     3061 agctgacatt catcccaggt ggcacttttc ggggaaatgt gcgcgcccgc gttcctgctg
     3121 gcgctgggcc tgtttctggc gctggacttc ccgctgttcc gtcagcagct tttcgcccac
     3181 ggccttgatg atcgcggcgg ccttggcctg catatcccga ttcaacggcc ccagggcgtc
     3241 cagaacgggc ttcaggcgct cccgaaggtc tcgggccgtc tcttgggctt gatcggcctt
     3301 cttgcgcatc tcacgcgctc ctgcggcggc ctgtagggca ggctcatacc cctgccgaac
     3361 cgcttttgtc agccggtcgg ccacggcttc cggcgtctca acgcgctttg agattcccag
     3421 cttttcggcc aatccctgcg gtgcataggc gcgtggctcg accgcttgcg ggctgatggt
     3481 gacgtggccc actggtggcc gctccagggc ctcgtagaac gcctgaatgc gcgtgtgacg
     3541 tgccttgctg ccctcgatgc cccgttgcag ccctagatcg gccacagcgg ccgcaaacgt
     3601 ggtctggtcg cgggtcatct gcgctttgtt gccgatgaac tccttggccg acagcctgcc
     3661 gtcctgcgtc agcggcacca cgaacgcggt catgtgcggg ctggtttcgt cacggtggat
     3721 gctggccgtc acgatgcgat ccgccccgta cttgtccgcc agccacttgt gcgccttctc
     3781 gaagaacgcc gcctgctgtt cttggctggc cgacttccac cattccgggc tggccgtcat
     3841 gacgtactcg accgccaaca cagcgtcctt gcgccgcttc tctggcagca actcgcgcag
     3901 tcggcccatc gcttcatcgg tgctgctggc cgcccagtgc tcgttctctg gcgtcctgct
     3961 ggcgtcagcg ttgggcgtct cgcgctcgcg gtaggcgtgc ttgagactgg ccgccacgtt
     4021 gcccattttc gccagcttct tgcatcgcat gatcgcgtat gccgccatgc ctgcccctcc
     4081 cttttggtgt ccaaccggct cgacgggggc agcgcaaggc ggtgcctccg gcgggccact
     4141 caatgcttga gtatactcac tagactttgc ttcgcaaagt cgtgaccgcc tacggcggct
     4201 gcggcgccct acgggcttgc tctccgggct tcgccctgcg cggtcgctgc gctcccttgc
     4261 cagcccgtgg atatgtggac gatggccgcg agcggccacc ggctggctcg cttcgctcgg
     4321 cccgtggaca accctgctgg acaagctgat ggacaggctg cgcctgccca cgagcttgac
     4381 cacagggatt gcccaccggc tacccagcct tcgaccacat acccaccggc tccaactgcg
     4441 cggcctgcgg ccttgcccca tcaatttttt taattttctc tggggaaaag cctccggcct
     4501 gcggcctgcg cgcttcgctt gccggttgga caccaagtgg aaggcgggtc aaggctcgcg
     4561 cagcgaccgc gcagcggctt ggccttgacg cgcctggaac gacccaagcc tatgcgagtg
     4621 ggggcagtcg aagggcgaag cccgcccgcc tgccccccga gcctcacggc ggcgagtgcg
     4681 ggggttccaa gggggcagcg ccaccttggg caaggccgaa ggccgcgcag tcgatcaaca
     4741 agccccggag gggccacttt ttgccggagg gggagccgcg ccgaaggcgt gggggaaccc
     4801 cgcaggggtg cccttctttg ggcaccaaag aactagatat agggcgaaat gcgaaagact
     4861 taaaaatcaa caacttaaaa aaggggggta cgcaacagct cattgcggca ccccccgcaa
     4921 tagctcattg cgtaggttaa agaaaatctg taattgactg ccacttttac gcaacgcata
     4981 attgttgtcg cgctgccgaa aagttgcagc tgattgcgca tggtgccgca accgtgcggc
     5041 acccctaccg catggagata agcatggcca cgcagtccag agaaatcggc attcaagcca
     5101 agaacaagcc cggtcactgg gtgcaaacgg aacgcaaagc gcatgaggcg tgggccgggc
     5161 ttattgcgag gaaacccacg gcggcaatgc tgctgcatca cctcgtggcg cagatgggcc
     5221 accagaacgc cgtggtggtc agccagaaga cactttccaa gctcatcgga cgttctttgc
     5281 ggacggtcca atacgcagtc aaggacttgg tggccgagcg ctggatctcc gtcgtgaagc
     5341 tcaacggccc cggcaccgtg tcggcctacg tggtcaatga ccgcgtggcg tggggccagc
     5401 cccgcgacca gttgcgcctg tcggtgttca gtgccgccgt ggtggttgat cacgacgacc
     5461 aggacgaatc gctgttgggg catggcgacc tgcgccgcat cccgaccctg tatccgggcg
     5521 agcagcaact accgaccggc cccggcgagg agccgcccag ccagcccggc attccgggca
     5581 tggaaccaga cctgccagcc ttgaccgaaa cggaggaatg ggaacggcgc gggcagcagc
     5641 gcctgccgat gcccgatgag ccgtgttttc tggacgatgg cgagccgttg gagccgccga
     5701 cacgggtcac gctgccgcgc cggtagcact tgggttgcgc agcaacccgt aagtgcgctg
     5761 ttccagacta tcggctgtag ccgcctcgcc gccctatacc ttgtctgcct ccccgcgttg
     5821 cgtcgcggtg catggagccg ggccacctcg acctgaatgg aagccggcgg cacctcgcta
     5881 acggattcac cgtttttatc aggctctggg aggcagaata aatgatcata tcgtcaatta
     5941 ttacctccac ggggagagcc tgagcaaact ggcctcaggc atttgagaag cacacggtca
     6001 cactgcttcc ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac
     6061 gaccctgccc tgaaccgacg accgggtcga atttgctttc gaatttctgc cattcatccg
     6121 cttattatca cttattcagg cgtagcaacc aggcgtttaa gggcaccaat aactgcctta
     6181 aaaaaattac gccccgccct gccactcatc gcagtacggc ctattggtta aaaaatgagc
     6241 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttccattc
     6301 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     6361 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     6421 ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact cactataggg
     6481 cgaattggag ctggggccgc aattcccaat tccccaggca tcaaataaaa cgaaaggctc
     6541 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
     6601 ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg
     6661 caggacgccc gccataaact gccaggaatt aattccccag gcatcaaata aaacgaaagg
     6721 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
     6781 gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc
     6841 gggcaggacg cccgccataa actgccagga attaattccc caggcatcaa ataaaacgaa
     6901 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     6961 tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt
     7021 ggcgggcagg acgcccgcca taaactgcca ggaattaatt ccccaggcat caaataaaac
     7081 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     7141 tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag
     7201 ggtggcgggc aggacgcccg ccataaactg ccaggaattg gggatcggat tcgagctaag
     7261 aattcccttg gggtatcttt aaatactgta gaaaagagga aggaaataat aaatggctaa
     7321 aatgagaata tcaccggaat tgaaaaaact gatcgaaaaa taccgctgcg taaaagatac
     7381 ggaaggaatg tctcctgcta aggtatataa gctggtggga gaaaatgaaa acctatattt
     7441 aaaaatgacg gacagccggt ataaagggac cacctatgat gtggaacggg aaaaggacat
     7501 gatgctatgg ctggaaggaa agctgcctgt tccaaaggtc ctgcactttg aacggcatga
     7561 tggctggagc aatctgctca tgagtgaggc cgatggcgtc ctttgctcgg aagagtatga
     7621 agatggacaa agccctgaaa agattatcga gctgtatgcg gagtgcatca ggctctttca
     7681 ctccatcgac atatcggatt gtccctatac gaatagctta gacagccgct tagccgaatt
     7741 ggattactta ctgaataacg atctggccga tgtggattgc gaaaactggg aagaagacac
     7801 tccatttaaa gatccgcgcg agctgtatga ttttttaaag acggaaaagc ccgaagagga
     7861 acttgtcttt tcccacggcg acctgggaga cagcaacatc tttgtgaaag atggcaaagt
     7921 aagtggcttt attgatcttg ggagaagcgg cagggcggac aagtggtatg acattgcctt
     7981 ctgcgtccgg tcgatcaggg aggatgtcgg ggaagaacag tatgtcgagc tattttttga
     8041 cttactgggg atcaagcctg attgggagaa aataaaatat tatattttac tggatgaatt
     8101 gttttagagc tcgcacgctg ccgcaagcac tcagggcgca agggctgcta aaggaagcgg
     8161 aacacgtaga aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt cagctactgg
     8221 gctatctgga caagggaaaa cgcaagcgca aagagaaagc aggtagcttg cagtgggctt
     8281 acatggcgat agctagactg ggcggtttta tggacagcaa gcgaaccgga attgccagct
     8341 ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttcttgccg
     8401 ccaaggatct gatggcgcag gggatcaaga tctgatcaag agacaggata ctagtggagg
     8461 aagaaaaaa
//
