LOCUS       sequence-226378-        7621 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 7621)
  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 7621)
  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: pMRE133
FEATURES             Location/Qualifiers
     source          1..7621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(1..3,4..6,7..717)
                     /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(46..67)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(307..326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     654..675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      738..832
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             876..1535
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(942..961)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1600..1686
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2025..2819)
                     /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(2136..2155)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2746..2765
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      4299..5070
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             5071..5733
                     /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     6427..6449
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6441..6458
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6442..6458
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6468..6487
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6468..6486
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      6529..6615
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6712..6798
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6895..6981
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7078..7164
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
       61 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      121 ggcaagctga ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc
      181 ctcgtgacca ccctgggcta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag
      241 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      301 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      361 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      421 aagctggagt acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac
      481 ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc
      541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      601 tacctgagct accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      661 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
      721 gtaattagct gagcttggac tcctgttgat agatccagta atgacctcag aactccatct
      781 ggatttgttc agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagcta
      841 gcttggcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata
      901 ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg
      961 ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa
     1021 agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg
     1081 ctcatccgga atttcgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc
     1141 acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat
     1201 accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg
     1261 aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc
     1321 cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc
     1381 ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga
     1441 ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac
     1501 aacagtactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt tattggtgcc
     1561 cttaaacgcc tggggtaatg actctctagc ttgaggcatc aaataaaacg aaaggctcag
     1621 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     1681 acaaatccgc cgctaggagc ttgcggccgc cagtgtgatg gatatctgca taaaaactgt
     1741 tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga
     1801 atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat gggggtgggc
     1861 gaagaactcc agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac
     1921 gattccgaag cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtgatggcag
     1981 gttgggcgtc gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca
     2041 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg
     2101 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg
     2161 tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
     2221 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
     2281 tcgggcatgc gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
     2341 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
     2401 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
     2461 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
     2521 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
     2581 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc
     2641 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
     2701 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
     2761 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg
     2821 cgaaacgatc ctcatcctgt ctcttgatca gatcttgatc ccctgcgcca tcagatcctt
     2881 ggcggcaaga aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc
     2941 ccagctggca attccggttc gcttgctgtc cataaaaccg cccagtctag ctatcgccat
     3001 gtaagcccac tgcaagctac ctgctttctc tttgcgcttg cgttttccct tgtccagata
     3061 gcccagtagc tgacattcat cccaggtggc acttttcggg gaaatgtgcg cgcccgcgtt
     3121 cctgctggcg ctgggcctgt ttctggcgct ggacttcccg ctgttccgtc agcagctttt
     3181 cgcccacggc cttgatgatc gcggcggcct tggcctgcat atcccgattc aacggcccca
     3241 gggcgtccag aacgggcttc aggcgctccc gaaggtctcg ggccgtctct tgggcttgat
     3301 cggccttctt gcgcatctca cgcgctcctg cggcggcctg tagggcaggc tcatacccct
     3361 gccgaaccgc ttttgtcagc cggtcggcca cggcttccgg cgtctcaacg cgctttgaga
     3421 ttcccagctt ttcggccaat ccctgcggtg cataggcgcg tggctcgacc gcttgcgggc
     3481 tgatggtgac gtggcccact ggtggccgct ccagggcctc gtagaacgcc tgaatgcgcg
     3541 tgtgacgtgc cttgctgccc tcgatgcccc gttgcagccc tagatcggcc acagcggccg
     3601 caaacgtggt ctggtcgcgg gtcatctgcg ctttgttgcc gatgaactcc ttggccgaca
     3661 gcctgccgtc ctgcgtcagc ggcaccacga acgcggtcat gtgcgggctg gtttcgtcac
     3721 ggtggatgct ggccgtcacg atgcgatccg ccccgtactt gtccgccagc cacttgtgcg
     3781 ccttctcgaa gaacgccgcc tgctgttctt ggctggccga cttccaccat tccgggctgg
     3841 ccgtcatgac gtactcgacc gccaacacag cgtccttgcg ccgcttctct ggcagcaact
     3901 cgcgcagtcg gcccatcgct tcatcggtgc tgctggccgc ccagtgctcg ttctctggcg
     3961 tcctgctggc gtcagcgttg ggcgtctcgc gctcgcggta ggcgtgcttg agactggccg
     4021 ccacgttgcc cattttcgcc agcttcttgc atcgcatgat cgcgtatgcc gccatgcctg
     4081 cccctccctt ttggtgtcca accggctcga cgggggcagc gcaaggcggt gcctccggcg
     4141 ggccactcaa tgcttgagta tactcactag actttgcttc gcaaagtcgt gaccgcctac
     4201 ggcggctgcg gcgccctacg ggcttgctct ccgggcttcg ccctgcgcgg tcgctgcgct
     4261 cccttgccag cccgtggata tgtggacgat ggccgcgagc ggccaccggc tggctcgctt
     4321 cgctcggccc gtggacaacc ctgctggaca agctgatgga caggctgcgc ctgcccacga
     4381 gcttgaccac agggattgcc caccggctac ccagccttcg accacatacc caccggctcc
     4441 aactgcgcgg cctgcggcct tgccccatca atttttttaa ttttctctgg ggaaaagcct
     4501 ccggcctgcg gcctgcgcgc ttcgcttgcc ggttggacac caagtggaag gcgggtcaag
     4561 gctcgcgcag cgaccgcgca gcggcttggc cttgacgcgc ctggaacgac ccaagcctat
     4621 gcgagtgggg gcagtcgaag ggcgaagccc gcccgcctgc cccccgagcc tcacggcggc
     4681 gagtgcgggg gttccaaggg ggcagcgcca ccttgggcaa ggccgaaggc cgcgcagtcg
     4741 atcaacaagc cccggagggg ccactttttg ccggaggggg agccgcgccg aaggcgtggg
     4801 ggaaccccgc aggggtgccc ttctttgggc accaaagaac tagatatagg gcgaaatgcg
     4861 aaagacttaa aaatcaacaa cttaaaaaag gggggtacgc aacagctcat tgcggcaccc
     4921 cccgcaatag ctcattgcgt aggttaaaga aaatctgtaa ttgactgcca cttttacgca
     4981 acgcataatt gttgtcgcgc tgccgaaaag ttgcagctga ttgcgcatgg tgccgcaacc
     5041 gtgcggcacc cctaccgcat ggagataagc atggccacgc agtccagaga aatcggcatt
     5101 caagccaaga acaagcccgg tcactgggtg caaacggaac gcaaagcgca tgaggcgtgg
     5161 gccgggctta ttgcgaggaa acccacggcg gcaatgctgc tgcatcacct cgtggcgcag
     5221 atgggccacc agaacgccgt ggtggtcagc cagaagacac tttccaagct catcggacgt
     5281 tctttgcgga cggtccaata cgcagtcaag gacttggtgg ccgagcgctg gatctccgtc
     5341 gtgaagctca acggccccgg caccgtgtcg gcctacgtgg tcaatgaccg cgtggcgtgg
     5401 ggccagcccc gcgaccagtt gcgcctgtcg gtgttcagtg ccgccgtggt ggttgatcac
     5461 gacgaccagg acgaatcgct gttggggcat ggcgacctgc gccgcatccc gaccctgtat
     5521 ccgggcgagc agcaactacc gaccggcccc ggcgaggagc cgcccagcca gcccggcatt
     5581 ccgggcatgg aaccagacct gccagccttg accgaaacgg aggaatggga acggcgcggg
     5641 cagcagcgcc tgccgatgcc cgatgagccg tgttttctgg acgatggcga gccgttggag
     5701 ccgccgacac gggtcacgct gccgcgccgg tagcacttgg gttgcgcagc aacccgtaag
     5761 tgcgctgttc cagactatcg gctgtagccg cctcgccgcc ctataccttg tctgcctccc
     5821 cgcgttgcgt cgcggtgcat ggagccgggc cacctcgacc tgaatggaag ccggcggcac
     5881 ctcgctaacg gattcaccgt ttttatcagg ctctgggagg cagaataaat gatcatatcg
     5941 tcaattatta cctccacggg gagagcctga gcaaactggc ctcaggcatt tgagaagcac
     6001 acggtcacac tgcttccggt agtcaataaa ccggtaaacc agcaatagac ataagcggct
     6061 atttaacgac cctgccctga accgacgacc gggtcgaatt tgctttcgaa tttctgccat
     6121 tcatccgctt attatcactt attcaggcgt agcaaccagg cgtttaaggg caccaataac
     6181 tgccttaaaa aaattacgcc ccgccctgcc actcatcgca gtacggccta ttggttaaaa
     6241 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     6301 tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     6361 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     6421 ggttttccca gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac
     6481 tatagggcga attggagctg gggccgcaat tcccaattcc ccaggcatca aataaaacga
     6541 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     6601 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
     6661 tggcgggcag gacgcccgcc ataaactgcc aggaattaat tccccaggca tcaaataaaa
     6721 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     6781 ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga
     6841 gggtggcggg caggacgccc gccataaact gccaggaatt aattccccag gcatcaaata
     6901 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     6961 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc
     7021 ggagggtggc gggcaggacg cccgccataa actgccagga attaattccc caggcatcaa
     7081 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     7141 aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg
     7201 cccggagggt ggcgggcagg acgcccgcca taaactgcca ggaattgggg atcggattcg
     7261 agctcgcacg ctgccgcaag cactcagggc gcaagggctg ctaaaggaag cggaacacgt
     7321 agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct
     7381 ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc
     7441 gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc
     7501 cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga
     7561 tctgatggcg caggggatca agatctgatc aagagacagg atactagtgg aggaagaaaa
     7621 a
//
