LOCUS       sequence-226742-        8449 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 8449)
  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 8449)
  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: pMRE146
FEATURES             Location/Qualifiers
     source          1..8449
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..735
                     /codon_start=1
                     /product="far-red fluorescent protein derived from 
                     mNeptune, with further red-shifted spectra"
                     /label=mCardinal
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTTEGEGKPYEGTQTQ
                     RIKVVEGGPLPFAFDILATCFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTVTQDTSLQDGCLIYNVKLRGVNFPSNGPVMQKKTLGWEATTETLYPADGGLEGRCD
                     MALKLVGGGHLHCNLKTTYRSKKPAKNLKMPGVYFVDRRLERIKEADNETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     terminator      752..846
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             890..1549
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(956..975)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1614..1700
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2039..2833)
                     /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(2150..2169)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2760..2779
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      4313..5084
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             5085..5747
                     /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     6441..6463
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6455..6472
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6456..6472
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6482..6501
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6482..6500
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      6543..6629
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6726..6812
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6909..6995
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7092..7178
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        join(7338..7343,7344..7360,7361..7366)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             7555..8088
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
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 cctccctagc aaactggggc acaaacttaa tggcatggac
      721 gagctgtaca agtaataatt agctgagctt ggactcctgt tgatagatcc agtaatgacc
      781 tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt
      841 gagaatccaa gctagcttgg cgagattttc aggagctaag gaagctaaaa tggagaaaaa
      901 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
      961 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
     1021 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
     1081 ccgcctgatg aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat
     1141 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
     1201 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt
     1261 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
     1321 cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
     1381 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct
     1441 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat
     1501 gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttttttaagg
     1561 cagttattgg tgcccttaaa cgcctggggt aatgactctc tagcttgagg catcaaataa
     1621 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     1681 ctctcctgag taggacaaat ccgccgctag gagcttgcgg ccgccagtgt gatggatatc
     1741 tgcataaaaa ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg
     1801 catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc
     1861 ccatgggggt gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg
     1921 gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa
     1981 tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc
     2041 agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga gcggcgatac
     2101 cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca atatcacggg
     2161 tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag tcgatgaatc
     2221 cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca tgggtcacga
     2281 cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg gctggcgcga
     2341 gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc atccgagtac
     2401 gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg
     2461 tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga gcaaggtgag
     2521 atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt cccgcttcag
     2581 tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg
     2641 ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg
     2701 ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt gtctgttgtg
     2761 cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg tgcaatccat
     2821 cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcagatctt gatcccctgc
     2881 gccatcagat ccttggcggc aagaaagcca tccagtttac tttgcagggc ttcccaacct
     2941 taccagaggg cgccccagct ggcaattccg gttcgcttgc tgtccataaa accgcccagt
     3001 ctagctatcg ccatgtaagc ccactgcaag ctacctgctt tctctttgcg cttgcgtttt
     3061 cccttgtcca gatagcccag tagctgacat tcatcccagg tggcactttt cggggaaatg
     3121 tgcgcgcccg cgttcctgct ggcgctgggc ctgtttctgg cgctggactt cccgctgttc
     3181 cgtcagcagc ttttcgccca cggccttgat gatcgcggcg gccttggcct gcatatcccg
     3241 attcaacggc cccagggcgt ccagaacggg cttcaggcgc tcccgaaggt ctcgggccgt
     3301 ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct cctgcggcgg cctgtagggc
     3361 aggctcatac ccctgccgaa ccgcttttgt cagccggtcg gccacggctt ccggcgtctc
     3421 aacgcgcttt gagattccca gcttttcggc caatccctgc ggtgcatagg cgcgtggctc
     3481 gaccgcttgc gggctgatgg tgacgtggcc cactggtggc cgctccaggg cctcgtagaa
     3541 cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg ccccgttgca gccctagatc
     3601 ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc tgcgctttgt tgccgatgaa
     3661 ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc acgaacgcgg tcatgtgcgg
     3721 gctggtttcg tcacggtgga tgctggccgt cacgatgcga tccgccccgt acttgtccgc
     3781 cagccacttg tgcgccttct cgaagaacgc cgcctgctgt tcttggctgg ccgacttcca
     3841 ccattccggg ctggccgtca tgacgtactc gaccgccaac acagcgtcct tgcgccgctt
     3901 ctctggcagc aactcgcgca gtcggcccat cgcttcatcg gtgctgctgg ccgcccagtg
     3961 ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc tcgcgctcgc ggtaggcgtg
     4021 cttgagactg gccgccacgt tgcccatttt cgccagcttc ttgcatcgca tgatcgcgta
     4081 tgccgccatg cctgcccctc ccttttggtg tccaaccggc tcgacggggg cagcgcaagg
     4141 cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg cttcgcaaag
     4201 tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc ttcgccctgc
     4261 gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc gagcggccac
     4321 cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga tggacaggct
     4381 gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc ttcgaccaca
     4441 tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt ttaattttct
     4501 ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg acaccaagtg
     4561 gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac gcgcctggaa
     4621 cgacccaagc ctatgcgagt gggggcagtc gaagggcgaa gcccgcccgc ctgccccccg
     4681 agcctcacgg cggcgagtgc gggggttcca agggggcagc gccaccttgg gcaaggccga
     4741 aggccgcgca gtcgatcaac aagccccgga ggggccactt tttgccggag ggggagccgc
     4801 gccgaaggcg tgggggaacc ccgcaggggt gcccttcttt gggcaccaaa gaactagata
     4861 tagggcgaaa tgcgaaagac ttaaaaatca acaacttaaa aaaggggggt acgcaacagc
     4921 tcattgcggc accccccgca atagctcatt gcgtaggtta aagaaaatct gtaattgact
     4981 gccactttta cgcaacgcat aattgttgtc gcgctgccga aaagttgcag ctgattgcgc
     5041 atggtgccgc aaccgtgcgg cacccctacc gcatggagat aagcatggcc acgcagtcca
     5101 gagaaatcgg cattcaagcc aagaacaagc ccggtcactg ggtgcaaacg gaacgcaaag
     5161 cgcatgaggc gtgggccggg cttattgcga ggaaacccac ggcggcaatg ctgctgcatc
     5221 acctcgtggc gcagatgggc caccagaacg ccgtggtggt cagccagaag acactttcca
     5281 agctcatcgg acgttctttg cggacggtcc aatacgcagt caaggacttg gtggccgagc
     5341 gctggatctc cgtcgtgaag ctcaacggcc ccggcaccgt gtcggcctac gtggtcaatg
     5401 accgcgtggc gtggggccag ccccgcgacc agttgcgcct gtcggtgttc agtgccgccg
     5461 tggtggttga tcacgacgac caggacgaat cgctgttggg gcatggcgac ctgcgccgca
     5521 tcccgaccct gtatccgggc gagcagcaac taccgaccgg ccccggcgag gagccgccca
     5581 gccagcccgg cattccgggc atggaaccag acctgccagc cttgaccgaa acggaggaat
     5641 gggaacggcg cgggcagcag cgcctgccga tgcccgatga gccgtgtttt ctggacgatg
     5701 gcgagccgtt ggagccgccg acacgggtca cgctgccgcg ccggtagcac ttgggttgcg
     5761 cagcaacccg taagtgcgct gttccagact atcggctgta gccgcctcgc cgccctatac
     5821 cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc gacctgaatg
     5881 gaagccggcg gcacctcgct aacggattca ccgtttttat caggctctgg gaggcagaat
     5941 aaatgatcat atcgtcaatt attacctcca cggggagagc ctgagcaaac tggcctcagg
     6001 catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta aaccagcaat
     6061 agacataagc ggctatttaa cgaccctgcc ctgaaccgac gaccgggtcg aatttgcttt
     6121 cgaatttctg ccattcatcc gcttattatc acttattcag gcgtagcaac caggcgttta
     6181 agggcaccaa taactgcctt aaaaaaatta cgccccgccc tgccactcat cgcagtacgg
     6241 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     6301 taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     6361 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     6421 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgagcgcgc
     6481 gtaatacgac tcactatagg gcgaattgga gctggggccg caattcccaa ttccccaggc
     6541 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     6601 cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc
     6661 aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggaat taattcccca
     6721 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     6781 tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga
     6841 agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg aattaattcc
     6901 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     6961 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg
     7021 cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggaattaat
     7081 tccccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     7141 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
     7201 ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggaatt
     7261 ggggatcgga ttcgagcttt agtacctaga tttagatgtc taaaaagcgg ggtaccgagg
     7321 acgcgtcaat tctcgaattg acataagcct gttcggttcg taaactgtaa tgcaagtagc
     7381 gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag
     7441 tggcggtttt catggcttgt tatgactgtt tttttgtaca gtctatgcct cgggcatcca
     7501 agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta
     7561 cgcagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt aggtggctca
     7621 agtatgggca tcattcgcac atgtaggctc ggccctgacc aagtcaaatc catgcgggct
     7681 gctcttgatc ttttcggtcg tgagttcgga gacgtagcca cctactccca acatcagccg
     7741 gactccgatt acctcgggaa cttgctccgt agtaagacat tcatcgcgct tgctgccttc
     7801 gaccaagaag cggttgttgg cgctctcgcg gcttacgttc tgcccaagtt tgagcagccg
     7861 cgtagtgaga tctatatcta tgatctcgca gtctccggcg agcaccggag gcagggcatt
     7921 gccaccgcgc tcatcaatct cctcaagcat gaggccaacg cgcttggtgc ttatgtgatc
     7981 tacgtgcaag cagattacgg tgacgatccc gcagtggctc tctatacaaa gttgggcata
     8041 cgggaagaag tgatgcactt tgatatcgac ccaagtaccg ccacctaaag ctcgcacgct
     8101 gccgcaagca ctcagggcgc aagggctgct aaaggaagcg gaacacgtag aaagccagtc
     8161 cgcagaaacg gtgctgaccc cggatgaatg tcagctactg ggctatctgg acaagggaaa
     8221 acgcaagcgc aaagagaaag caggtagctt gcagtgggct tacatggcga tagctagact
     8281 gggcggtttt atggacagca agcgaaccgg aattgccagc tggggcgccc tctggtaagg
     8341 ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc tgatggcgca
     8401 ggggatcaag atctgatcaa gagacaggat actagtggag gaagaaaaa
//
