LOCUS       40924_26286        4885 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pJM2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4885)
  AUTHORS   Murry JP, Pandey AK, Sassetti CM, Rubin EJ.
  TITLE     Phthiocerol Dimycocerosate Transport Is Required for Resisting 
            Interferon-gamma-Independent Immunity
  JOURNAL   J. Infect. Dis. 200 (5), 774-782 (2009)
  PUBMED    19622047
REFERENCE   2  (bases 1 to 4885)
  AUTHORS   Murry JP, Rubin EJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-AUG-2008) Microbiology and Molecular Genetics, Harvard
            Medical School, 200 Longwood Ave., Boston, MA 02115, USA
REFERENCE   3  (bases 1 to 4885)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4885)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. Infect. 
            Dis."; date: "2009"; volume: "200"; issue: "5"; pages: "774-782"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-AUG-2008) Microbiology and Molecular Genetics, Harvard Medical 
            School, 200 Longwood Ave., Boston, MA 02115, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     62..78
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        88..106
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(254..1111)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1112..1216)
                     /label=AmpR promoter
     rep_origin      complement(1243..1698)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     terminator      complement(1735..1782)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1849..1866)
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             complement(1916..1945)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="ASMTGGQQMG"
     protein_bind    complement(1954..1975)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(2263..2851)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        2980..3425
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             3426..4844
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
ORIGIN
        1 taaggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
       61 tgtaaaacga cggccagtga gcgcgcgtaa tacgactcac tatagggtct gacgctcagt
      121 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
      181 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
      241 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
      301 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
      361 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
      421 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
      481 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
      541 gtttgcgcaa cgttgttgcc attgctgcag gcatcgtggt gtcacgctcg tcgtttggta
      601 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
      661 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
      721 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
      781 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
      841 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
      901 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
      961 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     1021 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     1081 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     1141 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     1201 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgaaattgta aacgttaata
     1261 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     1321 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     1381 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     1441 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     1501 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     1561 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     1621 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     1681 cgccgctaca gggcgcgtcc cattcgccaa tccggatata gttcctcctt tcagcaaaaa
     1741 acccctcaag acccgtttag aggccccaag gggttatgct agttattgct cagcggtggc
     1801 agcagccaac tcagcttcct ttcgggcttt gttagcagcc ggatctcagt ggtggtggtg
     1861 gtggtgctcg agtgcggccg caagcttgtc gacggagctc gaattcggat ccgcgaccca
     1921 tttgctgtcc accagtcatg ctagccgtgc ctaatgagtg agctaactca cattaattgc
     1981 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     2041 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     2101 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2161 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2221 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2281 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2341 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2401 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2461 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2521 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2581 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2641 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2701 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2761 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2821 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2881 tgacgctcag tgcatatgga ggaatcactt cgcaatggcc aagacaattg cggatccagc
     2941 tgcaggtcga ctctagagga tcgatccttt ttaacccatc acatatacct gccgttcact
     3001 attatttagt gaaatgagat attatgatat tttctgaatt gtgattaaaa aggcaacttt
     3061 atgcccatgc aacagaaact ataaaaaata cagagaatga aaagaaacag atagattttt
     3121 tagttcttta ggcccgtagt ctgcaaatcc ttttatgatt ttctatcaaa caaaagagga
     3181 aaatagacca gttgcaatcc aaacgagagt ctaatagaat gaggtcgaaa agtaaatcgc
     3241 gcgggtttgt tactgataaa gcaggcaaga cctaaaatgt gtaaagggca aagtgtatac
     3301 tttggcgtca ccccttacat attttaggtc tttttttatt gtgcgtaact aacttgccat
     3361 cttcaaacag gagggctgga agaagcagac cgctaacaca gtacataaaa aaggagacat
     3421 gaacgatgaa catcaaaaag tttgcaaaac aagcaacagt attaaccttt actaccgcac
     3481 tgctggcagg aggcgcaact caagcgtttg cgaaagaaac gaaccaaaag ccatataagg
     3541 aaacatacgg catttcccat attacacgcc atgatatgct gcaaatccct gaacagcaaa
     3601 aaaatgaaaa atatcaagtt cctgaattcg attcgtccac aattaaaaat atctcttctg
     3661 caaaaggcct ggacgtttgg gacagctggc cattacaaaa cgctgacggc actgtcgcaa
     3721 actatcacgg ctaccacatc gtctttgcat tagccggaga tcctaaaaat gcggatgaca
     3781 catcgattta catgttctat caaaaagtcg gcgaaacttc tattgacagc tggaaaaacg
     3841 ctggccgcgt ctttaaagac agcgacaaat tcgatgcaaa tgattctatc ctaaaagacc
     3901 aaacacaaga atggtcaggt tcagccacat ttacatctga cggaaaaatc cgtttattct
     3961 acactgattt ctccggtaaa cattacggca aacaaacact gacaactgca caagttaacg
     4021 tatcagcatc agacagctct ttgaacatca acggtgtaga ggattataaa tcaatctttg
     4081 acggtgacgg aaaaacgtat caaaatgtac agcagttcat cgatgaaggc aactacagct
     4141 caggcgacaa ccatacgctg agagatcctc actacgtaga agataaaggc cacaaatact
     4201 tagtatttga agcaaacact ggaactgaag atggctacca aggcgaagaa tctttattta
     4261 acaaagcata ctatggcaaa agcacatcat tcttccgtca agaaagtcaa aaacttctgc
     4321 aaagcgataa aaaacgcacg gctgagttag caaacggcgc tctcggtatg attgagctaa
     4381 acgatgatta cacactgaaa aaagtgatga aaccgctgat tgcatctaac acagtaacag
     4441 atgaaattga acgcgcgaac gtctttaaaa tgaacggcaa atggtacctg ttcactgact
     4501 cccgcggatc aaaaatgacg attgacggca ttacgtctaa cgatatttac atgcttggtt
     4561 atgtttctaa ttctttaact ggcccataca agccgctgaa caaaactggc cttgtgttaa
     4621 aaatggatct tgatcctaac gatgtaacct ttacttactc acacttcgct gtacctcaag
     4681 cgaaaggaaa caatgtcgtg attacaagct atatgacaaa cagaggattc tacgcagaca
     4741 aacaatcaac gtttgcgcca agcttcctgc tgaacatcaa aggcaagaaa acatctgttg
     4801 tcaaagacag catccttgaa caaggacaat taacagttaa caaataaaaa cgcaaaagaa
     4861 aatgccgata tcctattggc ttaat
//
