LOCUS       sequence-226600-        8465 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 8465)
  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 8465)
  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: pMRE154
FEATURES             Location/Qualifiers
     source          1..8465
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..711
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     terminator      728..822
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             866..1525
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(932..951)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1590..1676
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2015..2809)
                     /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(2126..2145)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2736..2755
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      4289..5060
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             5061..5723
                     /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     6417..6439
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6431..6448
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6432..6448
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6458..6477
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6458..6476
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      6519..6605
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6702..6788
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6885..6971
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7068..7154
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             7310..8104
                     /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 gaataacatg gccatcatca aggagttcat gcgcttcaag
       61 gtgcgcatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
      121 cgcccctacg agggctttca gaccgctaag ctgaaggtga ccaagggtgg ccccctgccc
      181 ttcgcctggg acatcctgtc ccctcatttc acctacggct ccaaggccta cgtgaagcac
      241 cccgccgaca tccccgacta cttcaagctg tccttccccg agggcttcaa gtgggagcgc
      301 gtgatgaact acgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
      361 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgtg
      421 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggt
      481 gccctgaagg gcaagatcaa gatgaggctg aagctgaagg acggcggcca ctacacctcc
      541 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacatcgtc
      601 gacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
      661 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta ataattagct
      721 gagcttggac tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc
      781 agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagcta gcttggcgag
      841 attttcagga gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga
      901 tatatcccaa tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac
      961 ctataaccag accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa
     1021 gcacaagttt tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga
     1081 atttcgtatg gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta
     1141 caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga
     1201 tttccggcag tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc
     1261 ctatttccct aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag
     1321 tttcaccagt tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac
     1381 catgggcaaa tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca
     1441 tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg
     1501 cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt tattggtgcc cttaaacgcc
     1561 tggggtaatg actctctagc ttgaggcatc aaataaaacg aaaggctcag tcgaaagact
     1621 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     1681 cgctaggagc ttgcggccgc cagtgtgatg gatatctgca taaaaactgt tgtaattcat
     1741 taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg
     1801 gcatcagcac cttgtcgcct tgcgtataat atttgcccat gggggtgggc gaagaactcc
     1861 agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag
     1921 cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc
     1981 gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat
     2041 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
     2101 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
     2161 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
     2221 tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc
     2281 gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat
     2341 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg
     2401 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag
     2461 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca
     2521 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc
     2581 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca
     2641 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga
     2701 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct
     2761 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc
     2821 ctcatcctgt ctcttgatca gatcttgatc ccctgcgcca tcagatcctt ggcggcaaga
     2881 aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca
     2941 attccggttc gcttgctgtc cataaaaccg cccagtctag ctatcgccat gtaagcccac
     3001 tgcaagctac ctgctttctc tttgcgcttg cgttttccct tgtccagata gcccagtagc
     3061 tgacattcat cccaggtggc acttttcggg gaaatgtgcg cgcccgcgtt cctgctggcg
     3121 ctgggcctgt ttctggcgct ggacttcccg ctgttccgtc agcagctttt cgcccacggc
     3181 cttgatgatc gcggcggcct tggcctgcat atcccgattc aacggcccca gggcgtccag
     3241 aacgggcttc aggcgctccc gaaggtctcg ggccgtctct tgggcttgat cggccttctt
     3301 gcgcatctca cgcgctcctg cggcggcctg tagggcaggc tcatacccct gccgaaccgc
     3361 ttttgtcagc cggtcggcca cggcttccgg cgtctcaacg cgctttgaga ttcccagctt
     3421 ttcggccaat ccctgcggtg cataggcgcg tggctcgacc gcttgcgggc tgatggtgac
     3481 gtggcccact ggtggccgct ccagggcctc gtagaacgcc tgaatgcgcg tgtgacgtgc
     3541 cttgctgccc tcgatgcccc gttgcagccc tagatcggcc acagcggccg caaacgtggt
     3601 ctggtcgcgg gtcatctgcg ctttgttgcc gatgaactcc ttggccgaca gcctgccgtc
     3661 ctgcgtcagc ggcaccacga acgcggtcat gtgcgggctg gtttcgtcac ggtggatgct
     3721 ggccgtcacg atgcgatccg ccccgtactt gtccgccagc cacttgtgcg ccttctcgaa
     3781 gaacgccgcc tgctgttctt ggctggccga cttccaccat tccgggctgg ccgtcatgac
     3841 gtactcgacc gccaacacag cgtccttgcg ccgcttctct ggcagcaact cgcgcagtcg
     3901 gcccatcgct tcatcggtgc tgctggccgc ccagtgctcg ttctctggcg tcctgctggc
     3961 gtcagcgttg ggcgtctcgc gctcgcggta ggcgtgcttg agactggccg ccacgttgcc
     4021 cattttcgcc agcttcttgc atcgcatgat cgcgtatgcc gccatgcctg cccctccctt
     4081 ttggtgtcca accggctcga cgggggcagc gcaaggcggt gcctccggcg ggccactcaa
     4141 tgcttgagta tactcactag actttgcttc gcaaagtcgt gaccgcctac ggcggctgcg
     4201 gcgccctacg ggcttgctct ccgggcttcg ccctgcgcgg tcgctgcgct cccttgccag
     4261 cccgtggata tgtggacgat ggccgcgagc ggccaccggc tggctcgctt cgctcggccc
     4321 gtggacaacc ctgctggaca agctgatgga caggctgcgc ctgcccacga gcttgaccac
     4381 agggattgcc caccggctac ccagccttcg accacatacc caccggctcc aactgcgcgg
     4441 cctgcggcct tgccccatca atttttttaa ttttctctgg ggaaaagcct ccggcctgcg
     4501 gcctgcgcgc ttcgcttgcc ggttggacac caagtggaag gcgggtcaag gctcgcgcag
     4561 cgaccgcgca gcggcttggc cttgacgcgc ctggaacgac ccaagcctat gcgagtgggg
     4621 gcagtcgaag ggcgaagccc gcccgcctgc cccccgagcc tcacggcggc gagtgcgggg
     4681 gttccaaggg ggcagcgcca ccttgggcaa ggccgaaggc cgcgcagtcg atcaacaagc
     4741 cccggagggg ccactttttg ccggaggggg agccgcgccg aaggcgtggg ggaaccccgc
     4801 aggggtgccc ttctttgggc accaaagaac tagatatagg gcgaaatgcg aaagacttaa
     4861 aaatcaacaa cttaaaaaag gggggtacgc aacagctcat tgcggcaccc cccgcaatag
     4921 ctcattgcgt aggttaaaga aaatctgtaa ttgactgcca cttttacgca acgcataatt
     4981 gttgtcgcgc tgccgaaaag ttgcagctga ttgcgcatgg tgccgcaacc gtgcggcacc
     5041 cctaccgcat ggagataagc atggccacgc agtccagaga aatcggcatt caagccaaga
     5101 acaagcccgg tcactgggtg caaacggaac gcaaagcgca tgaggcgtgg gccgggctta
     5161 ttgcgaggaa acccacggcg gcaatgctgc tgcatcacct cgtggcgcag atgggccacc
     5221 agaacgccgt ggtggtcagc cagaagacac tttccaagct catcggacgt tctttgcgga
     5281 cggtccaata cgcagtcaag gacttggtgg ccgagcgctg gatctccgtc gtgaagctca
     5341 acggccccgg caccgtgtcg gcctacgtgg tcaatgaccg cgtggcgtgg ggccagcccc
     5401 gcgaccagtt gcgcctgtcg gtgttcagtg ccgccgtggt ggttgatcac gacgaccagg
     5461 acgaatcgct gttggggcat ggcgacctgc gccgcatccc gaccctgtat ccgggcgagc
     5521 agcaactacc gaccggcccc ggcgaggagc cgcccagcca gcccggcatt ccgggcatgg
     5581 aaccagacct gccagccttg accgaaacgg aggaatggga acggcgcggg cagcagcgcc
     5641 tgccgatgcc cgatgagccg tgttttctgg acgatggcga gccgttggag ccgccgacac
     5701 gggtcacgct gccgcgccgg tagcacttgg gttgcgcagc aacccgtaag tgcgctgttc
     5761 cagactatcg gctgtagccg cctcgccgcc ctataccttg tctgcctccc cgcgttgcgt
     5821 cgcggtgcat ggagccgggc cacctcgacc tgaatggaag ccggcggcac ctcgctaacg
     5881 gattcaccgt ttttatcagg ctctgggagg cagaataaat gatcatatcg tcaattatta
     5941 cctccacggg gagagcctga gcaaactggc ctcaggcatt tgagaagcac acggtcacac
     6001 tgcttccggt agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac
     6061 cctgccctga accgacgacc gggtcgaatt tgctttcgaa tttctgccat tcatccgctt
     6121 attatcactt attcaggcgt agcaaccagg cgtttaaggg caccaataac tgccttaaaa
     6181 aaattacgcc ccgccctgcc actcatcgca gtacggccta ttggttaaaa aatgagctga
     6241 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt tccattcgcc
     6301 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     6361 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     6421 gtcacgacgt tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggcga
     6481 attggagctg gggccgcaat tcccaattcc ccaggcatca aataaaacga aaggctcagt
     6541 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     6601 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
     6661 gacgcccgcc ataaactgcc aggaattaat tccccaggca tcaaataaaa cgaaaggctc
     6721 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
     6781 ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg
     6841 caggacgccc gccataaact gccaggaatt aattccccag gcatcaaata aaacgaaagg
     6901 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
     6961 gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc
     7021 gggcaggacg cccgccataa actgccagga attaattccc caggcatcaa ataaaacgaa
     7081 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     7141 tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt
     7201 ggcgggcagg acgcccgcca taaactgcca ggaattgggg atcggattcg agctaagaat
     7261 tcccttgggg tatctttaaa tactgtagaa aagaggaagg aaataataaa tggctaaaat
     7321 gagaatatca ccggaattga aaaaactgat cgaaaaatac cgctgcgtaa aagatacgga
     7381 aggaatgtct cctgctaagg tatataagct ggtgggagaa aatgaaaacc tatatttaaa
     7441 aatgacggac agccggtata aagggaccac ctatgatgtg gaacgggaaa aggacatgat
     7501 gctatggctg gaaggaaagc tgcctgttcc aaaggtcctg cactttgaac ggcatgatgg
     7561 ctggagcaat ctgctcatga gtgaggccga tggcgtcctt tgctcggaag agtatgaaga
     7621 tggacaaagc cctgaaaaga ttatcgagct gtatgcggag tgcatcaggc tctttcactc
     7681 catcgacata tcggattgtc cctatacgaa tagcttagac agccgcttag ccgaattgga
     7741 ttacttactg aataacgatc tggccgatgt ggattgcgaa aactgggaag aagacactcc
     7801 atttaaagat ccgcgcgagc tgtatgattt tttaaagacg gaaaagcccg aagaggaact
     7861 tgtcttttcc cacggcgacc tgggagacag caacatcttt gtgaaagatg gcaaagtaag
     7921 tggctttatt gatcttggga gaagcggcag ggcggacaag tggtatgaca ttgccttctg
     7981 cgtccggtcg atcagggagg atgtcgggga agaacagtat gtcgagctat tttttgactt
     8041 actggggatc aagcctgatt gggagaaaat aaaatattat attttactgg atgaattgtt
     8101 ttagagctcg cacgctgccg caagcactca gggcgcaagg gctgctaaag gaagcggaac
     8161 acgtagaaag ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag ctactgggct
     8221 atctggacaa gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag tgggcttaca
     8281 tggcgatagc tagactgggc ggttttatgg acagcaagcg aaccggaatt gccagctggg
     8341 gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca
     8401 aggatctgat ggcgcagggg atcaagatct gatcaagaga caggatacta gtggaggaag
     8461 aaaaa
//
