LOCUS       sequence-226433-        8429 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 8429)
  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 8429)
  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: pMRE141
FEATURES             Location/Qualifiers
     source          1..8429
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..23,24..29)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             218..751
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     CDS             join(1114..1116,1117..1830)
                     /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(1156..1177)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1417..1436)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1764..1785
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1844..1938
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1982..2641
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2048..2067)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2706..2792
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3131..3925)
                     /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(3242..3261)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3852..3871
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      5405..6176
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             6177..6839
                     /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     7533..7555
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7547..7564
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7548..7564
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7574..7593
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7574..7592
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      7635..7721
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7818..7904
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      8001..8087
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      8184..8270
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttgacataag cctgttcggt tcgtaaactg taatgcaagt agcgtatgcg ctcacgcaac
       61 tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct
      121 tgttatgact gtttttttgt acagtctatg cctcgggcat ccaagcagca agcgcgttac
      181 gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca gcaacgatgt
      241 tacgcagcag ggcagtcgcc ctaaaacaaa gttaggtggc tcaagtatgg gcatcattcg
      301 cacatgtagg ctcggccctg accaagtcaa atccatgcgg gctgctcttg atcttttcgg
      361 tcgtgagttc ggagacgtag ccacctactc ccaacatcag ccggactccg attacctcgg
      421 gaacttgctc cgtagtaaga cattcatcgc gcttgctgcc ttcgaccaag aagcggttgt
      481 tggcgctctc gcggcttacg ttctgcccaa gtttgagcag ccgcgtagtg agatctatat
      541 ctatgatctc gcagtctccg gcgagcaccg gaggcagggc attgccaccg cgctcatcaa
      601 tctcctcaag catgaggcca acgcgcttgg tgcttatgtg atctacgtgc aagcagatta
      661 cggtgacgat cccgcagtgg ctctctatac aaagttgggc atacgggaag aagtgatgca
      721 ctttgatatc gacccaagta ccgccaccta aagctcgcac gctgccgcaa gcactcaggg
      781 cgcaagggct gctaaaggaa gcggaacacg tagaaagcca gtccgcagaa acggtgctga
      841 ccccggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga
      901 aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt tttatggaca
      961 gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa gccctgcaaa
     1021 gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc aagatctgat
     1081 caagagacag gatactagtg gaggaagaaa aaaatgagca agggcgagga gctgttcacc
     1141 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1201 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     1261 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgtcctg gggcgtgcag
     1321 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1381 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1441 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1501 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactactt tagcgacaac
     1561 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac
     1621 aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1681 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa
     1741 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1801 actctcggca tggacgagct gtacaagtaa ttagctgagc ttggactcct gttgatagat
     1861 ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg
     1921 ttttttattg gtgagaatcc aagctagctt ggcgagattt tcaggagcta aggaagctaa
     1981 aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga
     2041 acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga
     2101 tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat
     2161 tcacattctt gcccgcctga tgaatgctca tccggaattt cgtatggcaa tgaaagacgg
     2221 tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga
     2281 aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata
     2341 ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga
     2401 gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt
     2461 ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg
     2521 cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca
     2581 tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta
     2641 atttttttaa ggcagttatt ggtgccctta aacgcctggg gtaatgactc tctagcttga
     2701 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     2761 tgtcggtgaa cgctctcctg agtaggacaa atccgccgct aggagcttgc ggccgccagt
     2821 gtgatggata tctgcataaa aactgttgta attcattaag cattctgccg acatggaagc
     2881 catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg
     2941 tataatattt gcccatgggg gtgggcgaag aactccagca tgagatcccc gcgctggagg
     3001 atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg
     3061 gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc
     3121 agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
     3181 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
     3241 caatatcacg ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac
     3301 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
     3361 catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt
     3421 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     3481 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     3541 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     3601 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     3661 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     3721 acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga
     3781 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     3841 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     3901 cgtgcaatcc atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcagatc
     3961 ttgatcccct gcgccatcag atccttggcg gcaagaaagc catccagttt actttgcagg
     4021 gcttcccaac cttaccagag ggcgccccag ctggcaattc cggttcgctt gctgtccata
     4081 aaaccgccca gtctagctat cgccatgtaa gcccactgca agctacctgc tttctctttg
     4141 cgcttgcgtt ttcccttgtc cagatagccc agtagctgac attcatccca ggtggcactt
     4201 ttcggggaaa tgtgcgcgcc cgcgttcctg ctggcgctgg gcctgtttct ggcgctggac
     4261 ttcccgctgt tccgtcagca gcttttcgcc cacggccttg atgatcgcgg cggccttggc
     4321 ctgcatatcc cgattcaacg gccccagggc gtccagaacg ggcttcaggc gctcccgaag
     4381 gtctcgggcc gtctcttggg cttgatcggc cttcttgcgc atctcacgcg ctcctgcggc
     4441 ggcctgtagg gcaggctcat acccctgccg aaccgctttt gtcagccggt cggccacggc
     4501 ttccggcgtc tcaacgcgct ttgagattcc cagcttttcg gccaatccct gcggtgcata
     4561 ggcgcgtggc tcgaccgctt gcgggctgat ggtgacgtgg cccactggtg gccgctccag
     4621 ggcctcgtag aacgcctgaa tgcgcgtgtg acgtgccttg ctgccctcga tgccccgttg
     4681 cagccctaga tcggccacag cggccgcaaa cgtggtctgg tcgcgggtca tctgcgcttt
     4741 gttgccgatg aactccttgg ccgacagcct gccgtcctgc gtcagcggca ccacgaacgc
     4801 ggtcatgtgc gggctggttt cgtcacggtg gatgctggcc gtcacgatgc gatccgcccc
     4861 gtacttgtcc gccagccact tgtgcgcctt ctcgaagaac gccgcctgct gttcttggct
     4921 ggccgacttc caccattccg ggctggccgt catgacgtac tcgaccgcca acacagcgtc
     4981 cttgcgccgc ttctctggca gcaactcgcg cagtcggccc atcgcttcat cggtgctgct
     5041 ggccgcccag tgctcgttct ctggcgtcct gctggcgtca gcgttgggcg tctcgcgctc
     5101 gcggtaggcg tgcttgagac tggccgccac gttgcccatt ttcgccagct tcttgcatcg
     5161 catgatcgcg tatgccgcca tgcctgcccc tcccttttgg tgtccaaccg gctcgacggg
     5221 ggcagcgcaa ggcggtgcct ccggcgggcc actcaatgct tgagtatact cactagactt
     5281 tgcttcgcaa agtcgtgacc gcctacggcg gctgcggcgc cctacgggct tgctctccgg
     5341 gcttcgccct gcgcggtcgc tgcgctccct tgccagcccg tggatatgtg gacgatggcc
     5401 gcgagcggcc accggctggc tcgcttcgct cggcccgtgg acaaccctgc tggacaagct
     5461 gatggacagg ctgcgcctgc ccacgagctt gaccacaggg attgcccacc ggctacccag
     5521 ccttcgacca catacccacc ggctccaact gcgcggcctg cggccttgcc ccatcaattt
     5581 ttttaatttt ctctggggaa aagcctccgg cctgcggcct gcgcgcttcg cttgccggtt
     5641 ggacaccaag tggaaggcgg gtcaaggctc gcgcagcgac cgcgcagcgg cttggccttg
     5701 acgcgcctgg aacgacccaa gcctatgcga gtgggggcag tcgaagggcg aagcccgccc
     5761 gcctgccccc cgagcctcac ggcggcgagt gcgggggttc caagggggca gcgccacctt
     5821 gggcaaggcc gaaggccgcg cagtcgatca acaagccccg gaggggccac tttttgccgg
     5881 agggggagcc gcgccgaagg cgtgggggaa ccccgcaggg gtgcccttct ttgggcacca
     5941 aagaactaga tatagggcga aatgcgaaag acttaaaaat caacaactta aaaaaggggg
     6001 gtacgcaaca gctcattgcg gcaccccccg caatagctca ttgcgtaggt taaagaaaat
     6061 ctgtaattga ctgccacttt tacgcaacgc ataattgttg tcgcgctgcc gaaaagttgc
     6121 agctgattgc gcatggtgcc gcaaccgtgc ggcaccccta ccgcatggag ataagcatgg
     6181 ccacgcagtc cagagaaatc ggcattcaag ccaagaacaa gcccggtcac tgggtgcaaa
     6241 cggaacgcaa agcgcatgag gcgtgggccg ggcttattgc gaggaaaccc acggcggcaa
     6301 tgctgctgca tcacctcgtg gcgcagatgg gccaccagaa cgccgtggtg gtcagccaga
     6361 agacactttc caagctcatc ggacgttctt tgcggacggt ccaatacgca gtcaaggact
     6421 tggtggccga gcgctggatc tccgtcgtga agctcaacgg ccccggcacc gtgtcggcct
     6481 acgtggtcaa tgaccgcgtg gcgtggggcc agccccgcga ccagttgcgc ctgtcggtgt
     6541 tcagtgccgc cgtggtggtt gatcacgacg accaggacga atcgctgttg gggcatggcg
     6601 acctgcgccg catcccgacc ctgtatccgg gcgagcagca actaccgacc ggccccggcg
     6661 aggagccgcc cagccagccc ggcattccgg gcatggaacc agacctgcca gccttgaccg
     6721 aaacggagga atgggaacgg cgcgggcagc agcgcctgcc gatgcccgat gagccgtgtt
     6781 ttctggacga tggcgagccg ttggagccgc cgacacgggt cacgctgccg cgccggtagc
     6841 acttgggttg cgcagcaacc cgtaagtgcg ctgttccaga ctatcggctg tagccgcctc
     6901 gccgccctat accttgtctg cctccccgcg ttgcgtcgcg gtgcatggag ccgggccacc
     6961 tcgacctgaa tggaagccgg cggcacctcg ctaacggatt caccgttttt atcaggctct
     7021 gggaggcaga ataaatgatc atatcgtcaa ttattacctc cacggggaga gcctgagcaa
     7081 actggcctca ggcatttgag aagcacacgg tcacactgct tccggtagtc aataaaccgg
     7141 taaaccagca atagacataa gcggctattt aacgaccctg ccctgaaccg acgaccgggt
     7201 cgaatttgct ttcgaatttc tgccattcat ccgcttatta tcacttattc aggcgtagca
     7261 accaggcgtt taagggcacc aataactgcc ttaaaaaaat tacgccccgc cctgccactc
     7321 atcgcagtac ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     7381 ttaacaaaat attaacgctt acaatttcca ttcgccattc aggctgcgca actgttggga
     7441 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     7501 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     7561 cagtgagcgc gcgtaatacg actcactata gggcgaattg gagctggggc cgcaattccc
     7621 aattccccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     7681 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
     7741 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccagga
     7801 attaattccc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     7861 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
     7921 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
     7981 ggaattaatt ccccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     8041 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg
     8101 atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg
     8161 ccaggaatta attccccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     8221 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag
     8281 cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa
     8341 ctgccaggaa ttggggatcg gattcgagct ttagtaccta gatttagatg tctaaaaagc
     8401 ggggtaccga ggacgcgtca attctcgaa
//
