LOCUS       sequence-226385-        7585 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 7585)
  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 7585)
  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: pMRE136
FEATURES             Location/Qualifiers
     source          1..7585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      702..796
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             840..1499
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(906..925)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1564..1650
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1989..2783)
                     /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(2100..2119)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2710..2729
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      4263..5034
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             5035..5697
                     /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     6391..6413
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6405..6422
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6406..6422
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6432..6451
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6432..6450
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      6493..6579
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6676..6762
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6859..6945
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7042..7128
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 atggtgagca agggcgagga gctgatcaag gagaacatgc acatgaagct gtacatggaa
       61 ggcaccgtga acaaccacca cttcaagtgc accaccgaag gggagggcaa gccctacgag
      121 ggcacccaga cccagaggat taaggtggtg gagggaggcc ccctgccgtt cgcattcgac
      181 atcctggcca cctgctttat gtacgggagc aagaccttca tcaaccacac ccagggcatc
      241 cccgatttct ttaagcagtc cttccctgag ggcttcacat gggagagagt caccacatac
      301 gaagacgggg gcgtgcttac cgttacccag gacaccagcc tccaggacgg ctgcttgatc
      361 tacaacgtca agctcagagg ggtgaacttc ccatccaacg gccctgtgat gcagaagaaa
      421 acactcggct gggaggccac caccgagacc ctgtaccccg ctgacggcgg cctggaaggc
      481 agatgcgaca tggccctgaa gctcgtgggc gggggccacc tgcactgcaa cctgaagacc
      541 acatacagat ccaagaaacc cgctaagaac ctcaagatgc ccggcgtcta ctttgtggac
      601 cgcagactgg aaagaatcaa ggaggccgac aatgagacct acgtcgagca gcacgaggtg
      661 gctgtggcca gatactgcga cctcctaatt agctgagctt ggactcctgt tgatagatcc
      721 agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt
      781 ttttattggt gagaatccaa gctagcttgg cgagattttc aggagctaag gaagctaaaa
      841 tggagaaaaa aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac
      901 attttgaggc atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata
      961 ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc
     1021 acattcttgc ccgcctgatg aatgctcatc cggaatttcg tatggcaatg aaagacggtg
     1081 agctggtgat atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa
     1141 cgttttcatc gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt
     1201 cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga
     1261 atatgttttt cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg
     1321 ccaatatgga caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg
     1381 acaaggtgct gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg
     1441 tcggcagaat gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat
     1501 ttttttaagg cagttattgg tgcccttaaa cgcctggggt aatgactctc tagcttgagg
     1561 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     1621 tcggtgaacg ctctcctgag taggacaaat ccgccgctag gagcttgcgg ccgccagtgt
     1681 gatggatatc tgcataaaaa ctgttgtaat tcattaagca ttctgccgac atggaagcca
     1741 tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta
     1801 taatatttgc ccatgggggt gggcgaagaa ctccagcatg agatccccgc gctggaggat
     1861 catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt
     1921 ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag
     1981 agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga
     2041 gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca
     2101 atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag
     2161 tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca
     2221 tgggtcacga cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg
     2281 gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc
     2341 atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc
     2401 ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga
     2461 gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt
     2521 cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac
     2581 gatagccgcg ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc ggtcttgaca
     2641 aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt
     2701 gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg
     2761 tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcagatctt
     2821 gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac tttgcagggc
     2881 ttcccaacct taccagaggg cgccccagct ggcaattccg gttcgcttgc tgtccataaa
     2941 accgcccagt ctagctatcg ccatgtaagc ccactgcaag ctacctgctt tctctttgcg
     3001 cttgcgtttt cccttgtcca gatagcccag tagctgacat tcatcccagg tggcactttt
     3061 cggggaaatg tgcgcgcccg cgttcctgct ggcgctgggc ctgtttctgg cgctggactt
     3121 cccgctgttc cgtcagcagc ttttcgccca cggccttgat gatcgcggcg gccttggcct
     3181 gcatatcccg attcaacggc cccagggcgt ccagaacggg cttcaggcgc tcccgaaggt
     3241 ctcgggccgt ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct cctgcggcgg
     3301 cctgtagggc aggctcatac ccctgccgaa ccgcttttgt cagccggtcg gccacggctt
     3361 ccggcgtctc aacgcgcttt gagattccca gcttttcggc caatccctgc ggtgcatagg
     3421 cgcgtggctc gaccgcttgc gggctgatgg tgacgtggcc cactggtggc cgctccaggg
     3481 cctcgtagaa cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg ccccgttgca
     3541 gccctagatc ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc tgcgctttgt
     3601 tgccgatgaa ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc acgaacgcgg
     3661 tcatgtgcgg gctggtttcg tcacggtgga tgctggccgt cacgatgcga tccgccccgt
     3721 acttgtccgc cagccacttg tgcgccttct cgaagaacgc cgcctgctgt tcttggctgg
     3781 ccgacttcca ccattccggg ctggccgtca tgacgtactc gaccgccaac acagcgtcct
     3841 tgcgccgctt ctctggcagc aactcgcgca gtcggcccat cgcttcatcg gtgctgctgg
     3901 ccgcccagtg ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc tcgcgctcgc
     3961 ggtaggcgtg cttgagactg gccgccacgt tgcccatttt cgccagcttc ttgcatcgca
     4021 tgatcgcgta tgccgccatg cctgcccctc ccttttggtg tccaaccggc tcgacggggg
     4081 cagcgcaagg cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg
     4141 cttcgcaaag tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc
     4201 ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc
     4261 gagcggccac cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga
     4321 tggacaggct gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc
     4381 ttcgaccaca tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt
     4441 ttaattttct ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg
     4501 acaccaagtg gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac
     4561 gcgcctggaa cgacccaagc ctatgcgagt gggggcagtc gaagggcgaa gcccgcccgc
     4621 ctgccccccg agcctcacgg cggcgagtgc gggggttcca agggggcagc gccaccttgg
     4681 gcaaggccga aggccgcgca gtcgatcaac aagccccgga ggggccactt tttgccggag
     4741 ggggagccgc gccgaaggcg tgggggaacc ccgcaggggt gcccttcttt gggcaccaaa
     4801 gaactagata tagggcgaaa tgcgaaagac ttaaaaatca acaacttaaa aaaggggggt
     4861 acgcaacagc tcattgcggc accccccgca atagctcatt gcgtaggtta aagaaaatct
     4921 gtaattgact gccactttta cgcaacgcat aattgttgtc gcgctgccga aaagttgcag
     4981 ctgattgcgc atggtgccgc aaccgtgcgg cacccctacc gcatggagat aagcatggcc
     5041 acgcagtcca gagaaatcgg cattcaagcc aagaacaagc ccggtcactg ggtgcaaacg
     5101 gaacgcaaag cgcatgaggc gtgggccggg cttattgcga ggaaacccac ggcggcaatg
     5161 ctgctgcatc acctcgtggc gcagatgggc caccagaacg ccgtggtggt cagccagaag
     5221 acactttcca agctcatcgg acgttctttg cggacggtcc aatacgcagt caaggacttg
     5281 gtggccgagc gctggatctc cgtcgtgaag ctcaacggcc ccggcaccgt gtcggcctac
     5341 gtggtcaatg accgcgtggc gtggggccag ccccgcgacc agttgcgcct gtcggtgttc
     5401 agtgccgccg tggtggttga tcacgacgac caggacgaat cgctgttggg gcatggcgac
     5461 ctgcgccgca tcccgaccct gtatccgggc gagcagcaac taccgaccgg ccccggcgag
     5521 gagccgccca gccagcccgg cattccgggc atggaaccag acctgccagc cttgaccgaa
     5581 acggaggaat gggaacggcg cgggcagcag cgcctgccga tgcccgatga gccgtgtttt
     5641 ctggacgatg gcgagccgtt ggagccgccg acacgggtca cgctgccgcg ccggtagcac
     5701 ttgggttgcg cagcaacccg taagtgcgct gttccagact atcggctgta gccgcctcgc
     5761 cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc
     5821 gacctgaatg gaagccggcg gcacctcgct aacggattca ccgtttttat caggctctgg
     5881 gaggcagaat aaatgatcat atcgtcaatt attacctcca cggggagagc ctgagcaaac
     5941 tggcctcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta
     6001 aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac gaccgggtcg
     6061 aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag gcgtagcaac
     6121 caggcgttta agggcaccaa taactgcctt aaaaaaatta cgccccgccc tgccactcat
     6181 cgcagtacgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     6241 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
     6301 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     6361 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     6421 gtgagcgcgc gtaatacgac tcactatagg gcgaattgga gctggggccg caattcccaa
     6481 ttccccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     6541 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
     6601 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggaat
     6661 taattcccca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     6721 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     6781 aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
     6841 aattaattcc ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     6901 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
     6961 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
     7021 aggaattaat tccccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     7081 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg
     7141 gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact
     7201 gccaggaatt ggggatcgga ttcgagctcg cacgctgccg caagcactca gggcgcaagg
     7261 gctgctaaag gaagcggaac acgtagaaag ccagtccgca gaaacggtgc tgaccccgga
     7321 tgaatgtcag ctactgggct atctggacaa gggaaaacgc aagcgcaaag agaaagcagg
     7381 tagcttgcag tgggcttaca tggcgatagc tagactgggc ggttttatgg acagcaagcg
     7441 aaccggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc aaagtaaact
     7501 ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagatct gatcaagaga
     7561 caggatacta gtggaggaag aaaaa
//
