LOCUS       pMRE132                7584 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  broad-host range plasmid to constitutively express fluorescent 
            protein genes in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    pMRE132
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7584)
  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 7584)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            //gt12365.cn
FEATURES             Location/Qualifiers
     source          1..7584
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             1..516
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIE"
     primer_bind     complement(43..64)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(304..323)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     651..672
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      701..795
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             839..1498
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(905..924)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1563..1649
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1988..2782)
                     /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(2099..2118)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2709..2728
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      4262..5033
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             5034..5696
                     /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     6390..6412
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6404..6421
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6405..6421
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6431..6450
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6431..6449
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      6492..6578
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6675..6761
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6858..6944
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7041..7127
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 atgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
       61 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
      121 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
      181 gtgaccaccc tgacctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag
      241 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
      301 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
      361 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
      421 ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc
      481 atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgccgac
      541 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
      601 ctgagcaccc agtccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg
      661 ctggagttcg tgaccgccgc cgggtaatta gctgagcttg gactcctgtt gatagatcca
      721 gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt
      781 tttattggtg agaatccaag ctagcttggc gagattttca ggagctaagg aagctaaaat
      841 ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca
      901 ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat
      961 tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca
     1021 cattcttgcc cgcctgatga atgctcatcc ggaatttcgt atggcaatga aagacggtga
     1081 gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac
     1141 gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc
     1201 gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa
     1261 tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc
     1321 caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga
     1381 caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt
     1441 cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaatt
     1501 tttttaaggc agttattggt gcccttaaac gcctggggta atgactctct agcttgaggc
     1561 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     1621 cggtgaacgc tctcctgagt aggacaaatc cgccgctagg agcttgcggc cgccagtgtg
     1681 atggatatct gcataaaaac tgttgtaatt cattaagcat tctgccgaca tggaagccat
     1741 cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat
     1801 aatatttgcc catgggggtg ggcgaagaac tccagcatga gatccccgcg ctggaggatc
     1861 atccagccgg cgtcccggaa aacgattccg aagcccaacc tttcatagaa ggcggcggtg
     1921 gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt cggtcatttc gaaccccaga
     1981 gtcccgctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag
     2041 cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc tcttcagcaa
     2101 tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc cggccacagt
     2161 cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag gcatcgccat
     2221 gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg
     2281 ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga ccggcttcca
     2341 tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg
     2401 gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc tcggcaggag
     2461 caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc cagtcccttc
     2521 ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg gccagccacg
     2581 atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa
     2641 aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag cagccgattg
     2701 tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga gaacctgcgt
     2761 gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc tgtctcttga tcagatcttg
     2821 atcccctgcg ccatcagatc cttggcggca agaaagccat ccagtttact ttgcagggct
     2881 tcccaacctt accagagggc gccccagctg gcaattccgg ttcgcttgct gtccataaaa
     2941 ccgcccagtc tagctatcgc catgtaagcc cactgcaagc tacctgcttt ctctttgcgc
     3001 ttgcgttttc ccttgtccag atagcccagt agctgacatt catcccaggt ggcacttttc
     3061 ggggaaatgt gcgcgcccgc gttcctgctg gcgctgggcc tgtttctggc gctggacttc
     3121 ccgctgttcc gtcagcagct tttcgcccac ggccttgatg atcgcggcgg ccttggcctg
     3181 catatcccga ttcaacggcc ccagggcgtc cagaacgggc ttcaggcgct cccgaaggtc
     3241 tcgggccgtc tcttgggctt gatcggcctt cttgcgcatc tcacgcgctc ctgcggcggc
     3301 ctgtagggca ggctcatacc cctgccgaac cgcttttgtc agccggtcgg ccacggcttc
     3361 cggcgtctca acgcgctttg agattcccag cttttcggcc aatccctgcg gtgcataggc
     3421 gcgtggctcg accgcttgcg ggctgatggt gacgtggccc actggtggcc gctccagggc
     3481 ctcgtagaac gcctgaatgc gcgtgtgacg tgccttgctg ccctcgatgc cccgttgcag
     3541 ccctagatcg gccacagcgg ccgcaaacgt ggtctggtcg cgggtcatct gcgctttgtt
     3601 gccgatgaac tccttggccg acagcctgcc gtcctgcgtc agcggcacca cgaacgcggt
     3661 catgtgcggg ctggtttcgt cacggtggat gctggccgtc acgatgcgat ccgccccgta
     3721 cttgtccgcc agccacttgt gcgccttctc gaagaacgcc gcctgctgtt cttggctggc
     3781 cgacttccac cattccgggc tggccgtcat gacgtactcg accgccaaca cagcgtcctt
     3841 gcgccgcttc tctggcagca actcgcgcag tcggcccatc gcttcatcgg tgctgctggc
     3901 cgcccagtgc tcgttctctg gcgtcctgct ggcgtcagcg ttgggcgtct cgcgctcgcg
     3961 gtaggcgtgc ttgagactgg ccgccacgtt gcccattttc gccagcttct tgcatcgcat
     4021 gatcgcgtat gccgccatgc ctgcccctcc cttttggtgt ccaaccggct cgacgggggc
     4081 agcgcaaggc ggtgcctccg gcgggccact caatgcttga gtatactcac tagactttgc
     4141 ttcgcaaagt cgtgaccgcc tacggcggct gcggcgccct acgggcttgc tctccgggct
     4201 tcgccctgcg cggtcgctgc gctcccttgc cagcccgtgg atatgtggac gatggccgcg
     4261 agcggccacc ggctggctcg cttcgctcgg cccgtggaca accctgctgg acaagctgat
     4321 ggacaggctg cgcctgccca cgagcttgac cacagggatt gcccaccggc tacccagcct
     4381 tcgaccacat acccaccggc tccaactgcg cggcctgcgg ccttgcccca tcaatttttt
     4441 taattttctc tggggaaaag cctccggcct gcggcctgcg cgcttcgctt gccggttgga
     4501 caccaagtgg aaggcgggtc aaggctcgcg cagcgaccgc gcagcggctt ggccttgacg
     4561 cgcctggaac gacccaagcc tatgcgagtg ggggcagtcg aagggcgaag cccgcccgcc
     4621 tgccccccga gcctcacggc ggcgagtgcg ggggttccaa gggggcagcg ccaccttggg
     4681 caaggccgaa ggccgcgcag tcgatcaaca agccccggag gggccacttt ttgccggagg
     4741 gggagccgcg ccgaaggcgt gggggaaccc cgcaggggtg cccttctttg ggcaccaaag
     4801 aactagatat agggcgaaat gcgaaagact taaaaatcaa caacttaaaa aaggggggta
     4861 cgcaacagct cattgcggca ccccccgcaa tagctcattg cgtaggttaa agaaaatctg
     4921 taattgactg ccacttttac gcaacgcata attgttgtcg cgctgccgaa aagttgcagc
     4981 tgattgcgca tggtgccgca accgtgcggc acccctaccg catggagata agcatggcca
     5041 cgcagtccag agaaatcggc attcaagcca agaacaagcc cggtcactgg gtgcaaacgg
     5101 aacgcaaagc gcatgaggcg tgggccgggc ttattgcgag gaaacccacg gcggcaatgc
     5161 tgctgcatca cctcgtggcg cagatgggcc accagaacgc cgtggtggtc agccagaaga
     5221 cactttccaa gctcatcgga cgttctttgc ggacggtcca atacgcagtc aaggacttgg
     5281 tggccgagcg ctggatctcc gtcgtgaagc tcaacggccc cggcaccgtg tcggcctacg
     5341 tggtcaatga ccgcgtggcg tggggccagc cccgcgacca gttgcgcctg tcggtgttca
     5401 gtgccgccgt ggtggttgat cacgacgacc aggacgaatc gctgttgggg catggcgacc
     5461 tgcgccgcat cccgaccctg tatccgggcg agcagcaact accgaccggc cccggcgagg
     5521 agccgcccag ccagcccggc attccgggca tggaaccaga cctgccagcc ttgaccgaaa
     5581 cggaggaatg ggaacggcgc gggcagcagc gcctgccgat gcccgatgag ccgtgttttc
     5641 tggacgatgg cgagccgttg gagccgccga cacgggtcac gctgccgcgc cggtagcact
     5701 tgggttgcgc agcaacccgt aagtgcgctg ttccagacta tcggctgtag ccgcctcgcc
     5761 gccctatacc ttgtctgcct ccccgcgttg cgtcgcggtg catggagccg ggccacctcg
     5821 acctgaatgg aagccggcgg cacctcgcta acggattcac cgtttttatc aggctctggg
     5881 aggcagaata aatgatcata tcgtcaatta ttacctccac ggggagagcc tgagcaaact
     5941 ggcctcaggc atttgagaag cacacggtca cactgcttcc ggtagtcaat aaaccggtaa
     6001 accagcaata gacataagcg gctatttaac gaccctgccc tgaaccgacg accgggtcga
     6061 atttgctttc gaatttctgc cattcatccg cttattatca cttattcagg cgtagcaacc
     6121 aggcgtttaa gggcaccaat aactgcctta aaaaaattac gccccgccct gccactcatc
     6181 gcagtacggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     6241 acaaaatatt aacgcttaca atttccattc gccattcagg ctgcgcaact gttgggaagg
     6301 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     6361 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     6421 tgagcgcgcg taatacgact cactataggg cgaattggag ctggggccgc aattcccaat
     6481 tccccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     6541 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
     6601 ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggaatt
     6661 aattccccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     6721 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
     6781 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccagga
     6841 attaattccc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     6901 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
     6961 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
     7021 ggaattaatt ccccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     7081 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg
     7141 atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg
     7201 ccaggaattg gggatcggat tcgagctcgc acgctgccgc aagcactcag ggcgcaaggg
     7261 ctgctaaagg aagcggaaca cgtagaaagc cagtccgcag aaacggtgct gaccccggat
     7321 gaatgtcagc tactgggcta tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt
     7381 agcttgcagt gggcttacat ggcgatagct agactgggcg gttttatgga cagcaagcga
     7441 accggaattg ccagctgggg cgccctctgg taaggttggg aagccctgca aagtaaactg
     7501 gatggctttc ttgccgccaa ggatctgatg gcgcagggga tcaagatctg atcaagagac
     7561 aggatactag tggaggaaga aaaa
//