LOCUS       Exported                7104 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMHC9-1, complete sequence.
ACCESSION   KF861545
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7104)
  AUTHORS   Maglennon GA, Cook BS, Deeney AS, Bosse JT, Peters SE, Langford PR, 
            Maskell DJ, Tucker AW, Wren BW, Rycroft AN.
  TITLE     Transposon mutagenesis in Mycoplasma hyopneumoniae using a novel 
            mariner-based system for generating random mutations
  JOURNAL   Vet. Res. 44 (1), 124 (2013)
  PUBMED    24359443
REFERENCE   2  (bases 1 to 7104)
  AUTHORS   Maglennon GA, Cook BS, Deeney AS, Bosse JT, Langford PR, Maskell DJ,
            Tucker AW, Wren BW, Rycroft AN.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-NOV-2013) Pathology & Pathogen Biology, Royal 
            Veterinary College, Hawkshead Lane, Hatfield, Hertfordshire AL9 7TA,
            United Kingdom
REFERENCE   3  (bases 1 to 7104)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7104
                     /organism="Cloning vector pMHC9-1"
                     /mol_type="other DNA"
                     /db_xref="taxon:1441630"
     CDS             805..1851
                     /codon_start=1
                     /product="transposase"
                     /label=transposase
                     /note="C9 mutant transposase from Himar1"
                     /protein_id="AHE63356.1"
                     /translation="MEKKEFRVLIKYCFLKGKNTVEAKTWLDNEFPDSAPGKSTIIDWY
                     AKFKRGEMSTEDGERSGRPKEVVTDENIKKIHKMILNDRKMKLIEIAEALKISKERVGH
                     IIHQYLDMRKLCAKWVPRELTFDQKQRRVDDSKRCLQLLTRNTPEFFRRYVTMDETWLH
                     HYTPESNRQSAEWTATGEPSPKRGKTQKSAGKVMASVFWDAHGIIFIDYLEKGKTINSD
                     YYMALLERLKVEIAAKRPHMKKKKVLFHQDNAPCHKSLRTMAKIHELGFELLPHPPYSP
                     DLAPSDFFLFSDIKRMLAGKKFGCNEEVIAETEAYFEAKPKEYYQNGIKKLEGRYNRCI
                     ALEGNYVE"
     CDS             2622..4541
                     /codon_start=1
                     /gene="tetM"
                     /product="tetracycline resistance protein"
                     /label=tetM
                     /protein_id="AHE63355.1"
                     /translation="MKIINIGVLAHVDAGKTTLTESLLYNSGAITELGSVDKGTTRTDN
                     TLLERQRGITIQTGITSFQWENTKVNIIDTPGHMDFLAEVYRSLSVLDGAILLISAKDG
                     VQAQTRILFHALRKMGIPTIFFINKIDQNGIDLSTVYQDIKEKLSAEIVIKQKVELYPN
                     VCVTNFTESEQWDTVIEGNDDLLEKYMSGKSLEALELEQEESIRFQNCSLFPLYHGSAK
                     SNIGIDNLIEVITNKFYSSTHRGPSELCGNVFKIEYTKKRQRLAYIRLYSGVLHLRDSV
                     RVSEKEKIKVTEMYTSINGELCKIDRAYSGEIVILQNEFLKLNSVLGDTKLLPQRKKIE
                     NPHPLLQTTVEPSKPEQREMLLDALLEISDSDPLLRYYVDSTTHEIILSFLGKVQMEVI
                     SALLQEKYHVEIEITEPTVIYMERPLKNAEYTIHIEVPPNPFWASIGLSVSPLPLGSGM
                     QYESSVSLGYLNQSFQNAVMEGIRYGCEQGLYGWNVTDCKICFKYGLYYSPVSTPADFR
                     MLAPIVLEQVLKKAGTELLEPYLSFKIYAPQEYLSRAYNDAPKYCANIVDTQLKNNEVI
                     LSGEIPARCIQEYRSDLTFFTNGRSVCLTELKGYHVTTGEPVCQPRRPNSRIDKVRYMF
                     NKIT"
     gene            2622..4541
                     /gene="tetM"
                     /label=tetM
     primer_bind     complement(4883..4899)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4907..4923
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4931..4961)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4976..4997
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5285..5873)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6044..6904)
                     /codon_start=1
                     /gene="bla"
                     /product="beta lactamase"
                     /label=bla
                     /protein_id="AHE63354.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(6044..6904)
                     /gene="bla"
                     /label=bla
     promoter        complement(6905..7009)
                     /gene="bla"
                     /label=AmpR promoter