LOCUS       40924_30805        7104 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMHC9-1, complete sequence.
ACCESSION   .
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   
REFERENCE   3  (bases 1 to 7104)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7104)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Vet. Res.";
            date: "2013"; volume: "44"; issue: "1"; pages: "124"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (16-NOV-2013) Pathology "
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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    complement(4907..4923)
                     /label=lac operator
                     /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    complement(4976..4997)
                     /label=CAP binding site
                     /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(6047..6904)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6905..7009)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgac ggggatttaa aacagaaact ccaaaaattg actatgcttt tattaaaaaa
      241 aaacagaaag aactctcact tttacctatt tttttaggat ttatttttgg atttttactt
      301 ttaattatta gtcttccttt tttaattcaa aattaataaa aaaatataga agaaaatttc
      361 ctagtaaagt tctaaaatta ttaaaaaaac agcattaaaa actgctgttt ttttaataat
      421 cctaatttta atattggatt atttatttaa attgttcaaa tttttttata ctaaaataaa
      481 ttataaaaga agactgatta gaaatttaga actattcaaa tctttcaaaa aagtggccta
      541 aaaacaatga taaaaaaatt tccaaacttt tttgttgcaa aaaaaaaaaa aaaaagtata
      601 attttaattg tacaagttaa ataaattttt cacttatctt tttttatttt gcaaaacttt
      661 taaaaaaatt agatatctaa attatattat atgattgaga aaatgaaaaa tttatttcct
      721 agaaccaaga taattgaggt ttaaaatatg agtaaaaaat caaaaacatt taaaattggt
      781 ttgactgccg gaattgttgg tgccatggaa aaaaaggaat ttcgtgtttt gataaaatac
      841 tgttttctga agggaaaaaa tacagtggaa gcaaaaactt ggcttgataa tgagtttccg
      901 gactctgccc cagggaaatc aacaataatt gattggtatg caaaattcaa gcgtggtgaa
      961 atgagcacgg aggacggtga acgcagtgga cgcccgaaag aggtggttac cgacgaaaac
     1021 atcaaaaaaa tccacaaaat gattttgaat gaccgtaaaa tgaagttgat cgagatagca
     1081 gaggccttaa agatatcaaa ggaacgtgtt ggtcatatca ttcatcaata tttggatatg
     1141 cggaagctct gtgcgaaatg ggtgccgcgc gagctcacat ttgaccaaaa acaacgacgt
     1201 gttgatgatt ctaagcggtg tttgcagctg ttaactcgta atacacccga gtttttccgt
     1261 cgatatgtga caatggatga aacatggctc catcactaca ctcctgagtc caatcgacag
     1321 tcggctgagt ggacagcgac cggtgaaccg tctccgaagc gtggaaagac tcaaaagtcc
     1381 gctggcaaag taatggcctc tgttttttgg gatgcgcatg gaataatttt tatcgattat
     1441 cttgagaagg gaaaaaccat caacagtgac tattatatgg cgttattgga gcgtttgaag
     1501 gtcgaaatcg cggcaaaacg gccccacatg aagaagaaaa aagtgttgtt ccaccaagac
     1561 aacgcaccgt gccacaagtc attgagaacg atggcaaaaa ttcatgaatt gggcttcgaa
     1621 ttgcttcccc acccgccgta ttctccagat ctggccccca gcgacttttt cttgttctca
     1681 gacatcaaaa ggatgctcgc agggaaaaaa tttggctgca atgaagaggt gatcgccgaa
     1741 actgaggcct attttgaggc aaaaccgaag gagtactacc aaaatggtat caaaaaattg
     1801 gaaggtcgtt ataatcgttg tatcgctctt gaagggaact atgttgaata atctcgagcg
     1861 gccgccagtg tgatggattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     1921 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     1981 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     2041 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     2101 tttaaagttc tgctaacagg ttggctgata agtccccggt ctgacacata gatggcgtcg
     2161 ctagtattaa atgcatatta tttttatata gtaccaacct ttcaaatgat tcccagatct
     2221 caaactggaa caacactcaa ccctatctcg gtctaagctt gcatgcctgc aggcagaatt
     2281 aaaagttagt gaacaagaaa acagtgaagc accagtttct gaaccaaaag aagacgaaaa
     2341 aacaaaaaaa gattaagcaa tttatttgga aaatcttttt ttgttttttt aagaaatatt
     2401 tattgttttt tttaaaaaat tattgtacag ttgctactat aagggaaaga aaaaaagaaa
     2461 gatataaatt gtataaagta gggttagaag caattaataa ttattattaa tgttattttt
     2521 ctcttatata ttcaatgtaa ttttaattac atttgctttt aataaaaaca ctacttaata
     2581 gagaaaggaa atataagatc ctttgaatgg aggaaaatca catgaaaatt attaatattg
     2641 gagttttagc tcatgttgat gcaggaaaaa ctaccttaac agaaagctta ttatataaca
     2701 gtggagcgat tacagaatta ggaagcgtgg acaaaggtac aacgaggacg gataatacgc
     2761 ttttagaacg tcagagagga attacaattc agacaggaat aacctctttt cagtgggaaa
     2821 atacgaaggt gaacatcata gacacgccag gacatatgga tttcttagca gaagtatatc
     2881 gttcattatc agttttagat ggggcaattc tactgatttc tgcaaaagat ggcgtacaag
     2941 cacaaactcg tatattattt catgcactta ggaaaatggg gattcccaca atctttttta
     3001 tcaataagat tgaccaaaat ggaattgatt tatcaacggt ttatcaggat attaaagaga
     3061 aactttctgc cgaaattgta atcaaacaga aggtagaact gtatcctaat gtgtgtgtga
     3121 cgaactttac cgaatctgaa caatgggata cggtaataga gggaaacgat gaccttttag
     3181 agaaatatat gtccggtaaa tcattagaag cattggaact cgaacaagag gaaagcataa
     3241 gatttcagaa ttgttctctg ttccctcttt atcatggaag tgcaaaaagt aatataggga
     3301 ttgataacct tatagaagtt attactaata aattttattc atcaacacat cgaggtccgt
     3361 ctgaactttg cggaaatgtt ttcaaaattg aatatacaaa aaaaagacaa cgtcttgcat
     3421 atatacgcct ttatagtgga gtactacatt tacgagattc ggttagagta tcagaaaagg
     3481 aaaaaataaa agttacagaa atgtatactt caataaatgg tgaattatgt aagattgata
     3541 gagcttattc tggagaaatt gttattttgc aaaatgagtt tttgaagtta aatagtgttc
     3601 ttggagatac aaaactattg ccacagagaa aaaagattga aaatccgcac cctctactac
     3661 aaacaactgt tgaaccgagt aaacctgaac agagagaaat gttgcttgat gcccttttgg
     3721 aaatctcaga tagtgatccg cttctacgat attacgtgga ttctacgaca catgaaatta
     3781 tactttcttt cttagggaaa gtacaaatgg aagtgattag tgcactgttg caagaaaagt
     3841 atcatgtgga gatagaaata acagagccta cagtcattta tatggagaga ccgttaaaaa
     3901 atgcagaata taccattcac atcgaagtgc cgccaaatcc tttctgggct tccattggtc
     3961 tatctgtatc accgcttccg ttgggaagtg gaatgcagta tgagagctcg gtttctcttg
     4021 gatacttaaa tcaatcattt caaaatgcag ttatggaagg gatacgctat ggttgtgaac
     4081 aaggattgta tggttggaat gtgacggact gtaaaatctg ttttaagtat ggcttatact
     4141 atagccctgt tagtacccca gcagattttc ggatgcttgc tcctattgta ttggaacaag
     4201 tcttaaaaaa agctggaaca gaattgttag agccatatct tagttttaaa atttatgcgc
     4261 cacaggaata tctttcacga gcatacaacg atgctcctaa atattgtgcg aacatcgtag
     4321 acactcaatt gaaaaataat gaggtcattc ttagtggaga aatccctgct cggtgtattc
     4381 aagaatatcg tagtgattta actttcttta caaatggacg tagtgtttgt ttaacagagt
     4441 taaaagggta ccatgttact accggtgaac ctgtttgcca gccccgtcgt ccaaatagtc
     4501 ggatagataa agtacgatat atgttcaata aaataactta gtgtaattta agttgttata
     4561 taaacgcgtc aattcgaggg gtatcgctct tgaagggaac tatgttgaat aataaaaacg
     4621 aattttcaca aaaaaatgtg tttttctttg ttagaccggg gacttatcag ccaacctgtt
     4681 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4741 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4801 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcgggtac
     4861 cgagctcgaa ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     4921 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     4981 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5041 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5101 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5161 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5221 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5281 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5341 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5401 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5461 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5521 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5581 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5641 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5701 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     5761 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5821 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5881 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5941 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6001 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6061 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6121 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6181 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6241 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6301 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6361 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6421 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6481 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6541 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6601 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6661 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6721 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6781 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6841 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6901 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6961 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7021 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     7081 ggcgtatcac gaggcccttt cgtc
//
