LOCUS       sequence-270239-        7119 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  rhamnose-inducible expression vector for replication in E. coli and 
            P. putida.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7119)
  AUTHORS   Jeske M, Altenbuchner J
  TITLE     The Escherichia coli rhamnose promoter rhaP(BAD) is in Pseudomonas 
            putida KT2440 independent of Crp-cAMP activation.
  JOURNAL   Appl Microbiol Biotechnol. 2010 Feb;85(6):1923-33. doi: 
            10.1007/s00253-009-2245-8. Epub 2009 Sep 30.
  PUBMED    19789867
REFERENCE   2  (bases 1 to 7119)
  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"; doi: 
            "10.1007/s00253-009-2245-8"; journalName: "Appl Microbiol 
            Biotechnol"; date: "2010-02"; volume: "85"; issue: "6"; pages: 
            "1923-33"
COMMENT     Sequence Label: pJeM1
FEATURES             Location/Qualifiers
     source          1..7119
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1023..1794
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1795..2457
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     CDS             complement(2764..3477)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVYGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     complement(2809..2830)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3158..3177
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3417..3438
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(3484..3501)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(3547..3665)
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     CDS             3811..4647
                     /codon_start=1
                     /gene="E. coli rhaS"
                     /product="positive regulator of the rhaB promoter
                     "
                     /label=rhaS
                     /note="activated by L-rhamnose"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     CDS             4721..5569
                     /codon_start=1
                     /gene="E. coli rhaR"
                     /product="transcriptional activator of rhaR and rhaS"
                     /label=rhaR
                     /note="needed for rhamnose-dependent activation of the rhaB
                     promoter"
                     /translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
                     RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI
                     PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR
                     YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN
                     QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK
                     D"
     CDS             complement(5868..6662)
                     /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(5979..5998)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6589..6608
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 ctcgggccgt ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct cctgcggcgg
       61 cctgtagggc aggctcatac ccctgccgaa ccgcttttgt cagccggtcg gccacggctt
      121 ccggcgtctc aacgcgcttt gagattccca gcttttcggc caatccctgc ggtgcatagg
      181 cgcgtggctc gaccgcttgc gggctgatgg tgacgtggcc cactggtggc cgctccaggg
      241 cctcgtagaa cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg ccccgttgca
      301 gccctagatc ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc tgcgctttgt
      361 tgccgatgaa ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc acgaacgcgg
      421 tcatgtgcgg gctggtttcg tcacggtgga tgctggccgt cacgatgcga tccgccccgt
      481 acttgtccgc cagccacttg tgcgccttct cgaagaacgc cgcctgctgt tcttggctgg
      541 ccgacttcca ccattccggg ctggccgtca tgacgtactc gaccgccaac acagcgtcct
      601 tgcgccgctt ctctggcagc aactcgcgca gtcggcccat cgcttcatcg gtgctgctgg
      661 ccgcccagtg ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc tcgcgctcgc
      721 ggtaggcgtg cttgagactg gccgccacgt tgcccatttt cgccagcttc ttgcatcgca
      781 tgatcgcgta tgccgccatg cctgcccctc ccttttggtg tccaaccggc tcgacggggg
      841 cagcgcaagg cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg
      901 cttcgcaaag tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc
      961 ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc
     1021 gagcggccac cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga
     1081 tggacaggct gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc
     1141 ttcgaccaca tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt
     1201 ttaattttct ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg
     1261 acaccaagtg gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac
     1321 gcgcctggaa cgacccaagc ctatgcgagt gggggcagtc gaagggcgaa gcccgcccgc
     1381 ctgccccccg agcctcacgg cggcgagtgc gggggttcca agggggcagc gccaccttgg
     1441 gcaaggccga aggccgcgca gtcgatcaac aagccccgga ggggccactt tttgccggag
     1501 ggggagccgc gccgaaggcg tgggggaacc ccgcaggggt gcccttcttt gggcaccaaa
     1561 gaactagata tagggcgaaa tgcgaaagac ttaaaaatca acaacttaaa aaaggggggt
     1621 acgcaacagc tcattgcggc accccccgca atagctcatt gcgtaggtta aagaaaatct
     1681 gtaattgact gccactttta cgcaacgcat aattgttgtc gcgctgccga aaagttgcag
     1741 ctgattgcgc atggtgccgc aaccgtgcgg cacccctacc gcatggagat aagcatggcc
     1801 acgcagtcca gagaaatcgg cattcaagcc aagaacaagc ccggtcactg ggtgcaaacg
     1861 gaacgcaaag cgcatgaggc gtgggccggg cttattgcga ggaaacccac ggcggcaatg
     1921 ctgctgcatc acctcgtggc gcagatgggc caccagaacg ccgtggtggt cagccagaag
     1981 acactttcca agctcatcgg acgttctttg cggacggtcc aatacgcagt caaggacttg
     2041 gtggccgagc gctggatctc cgtcgtgaag ctcaacggcc ccggcaccgt gtcggcctac
     2101 gtggtcaatg accgcgtggc gtggggccag ccccgcgacc agttgcgcct gtcggtgttc
     2161 agtgccgccg tggtggttga tcacgacgac caggacgaat cgctgttggg gcatggcgac
     2221 ctgcgccgca tcccgaccct gtatccgggc gagcagcaac taccgaccgg ccccggcgag
     2281 gagccgccca gccagcccgg cattccgggc atggaaccag acctgccagc cttgaccgaa
     2341 acggaggaat gggaacggcg cgggcagcag cgcctgccga tgcccgatga gccgtgtttt
     2401 ctggacgatg gcgagccgtt ggagccgccg acacgggtca cgctgccgcg ccggtagcac
     2461 ttgggttgcg cagcaacccg taagtgcgct gttccagact atcggctgta gccgcctcgc
     2521 cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc
     2581 gacctgaatg gaagccggcg gcacctcgct aacggattca ccgtttttat caggctctgg
     2641 gaggcagaat aaatgatcat atcgtcaatt attacctcca cggggagagc ctgagcaaac
     2701 tggcctcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta
     2761 agcttacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag
     2821 caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact gggtgctcag
     2881 gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg
     2941 gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat
     3001 gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt tgtactccag
     3061 cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt
     3121 caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt
     3181 gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg
     3241 ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca gggtggtcac
     3301 gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca gggtcagctt
     3361 gccgtaggtg gcatcgccct cgccctcgcc gtacacgctg aacttgtggc cgtttacgtc
     3421 gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctgga
     3481 tccatgatga tgatgatgat gcgtaatcat ggtcatatgt atatctcctt cttaagaatt
     3541 gttcattacg accagtctaa aaagcgcctg aattcgcgac cttctcgtta ctgacaggaa
     3601 aatgggccat tggcaaccag ggaaagatga acgtgatgat gttcacaatt tgctgaattg
     3661 tggtgatgtg atgctcaccg catttcctga aaattcacgc tgtatcttga aaaatcgacg
     3721 ttttttacgt ggttttccgt cgaaaattta aggtaagaac ctgacctcgt gattactatt
     3781 tcgccgtgtt gacgacatca ggaggccagt atgaccgtat tacatagtgt ggattttttt
     3841 ccgtctggta acgcgtccgt ggcgatagaa ccccggctcc cgcaggcgga ttttcctgaa
     3901 catcatcatg attttcatga aattgtgatt gtcgaacatg gcacgggtat tcatgtgttt
     3961 aatgggcagc cctataccat caccggtggc acggtctgtt tcgtacgcga tcatgatcgg
     4021 catctgtatg aacataccga taatctgtgt ctgaccaatg tgctgtatcg ctcgccggat
     4081 cgatttcagt ttctcgccgg gctgaatcag ttgctgccac aagagctgga tgggcagtat
     4141 ccgtctcact ggcgcgttaa ccacagcgta ttgcagcagg tgcgacagct ggttgcacag
     4201 atggaacagc aggaagggga aaatgattta ccctcgaccg ccagtcgcga gatcttgttt
     4261 atgcaattac tgctcttgct gcgtaaaagc agtttgcagg agaacctgga aaacagcgca
     4321 tcacgtctca acttgcttct ggcctggctg gaggaccatt ttgccgatga ggtgaattgg
     4381 gatgccgtgg cggatcaatt ttctctttca ctgcgtacgc tacatcggca gcttaagcag
     4441 caaacgggac tgacgcctca gcgatacctg aaccgcctgc gactgatgaa agcccgacat
     4501 ctgctacgcc acagcgaggc cagcgttact gacatcgcct atcgctgtgg attcagcgac
     4561 agtaaccact tttcgacgct ttttcgccga gagtttaact ggtcaccgcg tgatattcgc
     4621 cagggacggg atggctttct gcaataacgc gaatcttctc aacgtatttg tacgccatat
     4681 tgcgaataat caacttcgtt ctctggccga ggtagccacg gtggcgcatc agttaaaact
     4741 tctcaaagat gatttttttg ccagcgacca gcaggcagtc gctgtggctg accgttatcc
     4801 gcaagatgtc tttgctgaac atacacatga tttttgtgag ctggtgattg tctggcgcgg
     4861 taatggcctg catgtactca acgatcgccc ttatcgcatt acccgtggcg atctctttta
     4921 cattcatgct gacgataaac actcctacgc ttccgttaac gatctggttt tgcagaatat
     4981 tatttattgc ccggagcgtc tgaagctgaa tcttgactgg cagggggcga ttccgggatt
     5041 taacgccagc gcagggcaac cacactggcg cttaggtagc atggggatgg cgcaggcgcg
     5101 gcaggttatc ggtcagcttg agcatgaaag tagtcagcat gtgccgtttg ctaacgaaat
     5161 ggctgagttg ctgttcgggc agttggtgat gttgctgaat cgccatcgtt acaccagtga
     5221 ttcgttgccg ccaacatcca gcgaaacgtt gctggataag ctgattaccc ggctggcggc
     5281 tagcctgaaa agtccctttg cgctggataa attttgtgat gaggcatcgt gcagtgagcg
     5341 cgttttgcgt cagcaatttc gccagcagac tggaatgacc atcaatcaat atctgcgaca
     5401 ggtcagagtg tgtcatgcgc aatatcttct ccagcatagc cgcctgttaa tcagtgatat
     5461 ttcgaccgaa tgtggctttg aagatagtaa ctatttttcg gtggtgttta cccgggaaac
     5521 cgggatgacg cccagccagt ggcgtcatct caattcgcag aaagattaat gcataaaaac
     5581 tgttgtaatt cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc
     5641 tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catgggggtg
     5701 ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa
     5761 aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg
     5821 caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg
     5881 tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg
     5941 aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct
     6001 atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
     6061 ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg
     6121 ccgtcgggca tgcgcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc
     6181 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
     6241 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
     6301 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
     6361 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
     6421 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtcc
     6481 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
     6541 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
     6601 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
     6661 atgcgaaacg atcctcatcc tgtctcttga tcagatcttg atcccctgcg ccatcagatc
     6721 cttggcggca agaaagccat ccagtttact ttgcagggct tcccaacctt accagagggc
     6781 gccccagctg gcaattccgg ttcgcttgct gtccataaaa ccgcccagtc tagctatcgc
     6841 catgtaagcc cactgcaagc tacctgcttt ctctttgcgc ttgcgttttc ccttgtccag
     6901 atagcccagt agctgacatt catcccaggt ggcacttttc ggggaaatgt gcgcgcccgc
     6961 gttcctgctg gcgctgggcc tgtttctggc gctggacttc ccgctgttcc gtcagcagct
     7021 tttcgcccac ggccttgatg atcgcggcgg ccttggcctg catatcccga ttcaacggcc
     7081 ccagggcgtc cagaacgggc ttcaggcgct cccgaaggt
//
