LOCUS       Exported               13561 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  TnphoZ mutagenesis vector pMHL120, complete sequence.
ACCESSION   AY028776
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13561)
  AUTHORS   Lee MH, Nittayajarn A, Ross RP, Rothschild CB, Parsonage D, 
            Claiborne A, Rubens CE.
  TITLE     Characterization of Enterococcus faecalis alkaline phosphatase and 
            use in identifying Streptococcus agalactiae secreted proteins
  JOURNAL   J. Bacteriol. 181 (18), 5790-5799 (1999)
  PUBMED    10482522
REFERENCE   2  (bases 1 to 13561)
  AUTHORS   Clancy A, Lee MH, Jones AL, Rubens CE.
  TITLE     Construction and characterization of transposon TnphoZ for the 
            identification of genes encoding exported proteins in Streptococcus 
            agalactiae
  JOURNAL   FEMS Microbiol. Lett. 241 (2), 257-264 (2004)
  PUBMED    15598541
REFERENCE   3  (bases 1 to 13561)
  AUTHORS   Lee MH, Clancy A, Jones AL, Nittayajarn A, Rubens CE.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-MAR-2001) Infectious Disease, Childrens Hospital and 
            Regional Medical Center, 4800 Sand Point Way NE, Seattle, WA 98105, 
            USA
REFERENCE   4  (bases 1 to 13561)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..13561
                     /organism="TnphoZ mutagenesis vector pMHL120"
                     /lab_host="Streptococcus agalactiae"
                     /mol_type="genomic DNA"
                     /label=transposon TnphoZ delivery vehicle
                     /note="transposon TnphoZ delivery vehicle"
                     /db_xref="taxon:156762"
     RBS             3821..3829
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     CDS             complement(4167..4928)
                     /codon_start=1
                     /gene="erm"
                     /product="rRNA methylase"
                     /label=erm
                     /note="confers erythromycin-resistance"
                     /protein_id="AAK27828.1"
                     /translation="MYHFRSDYMNKNIKYSQNFLTNEKVLNQIIKQLNLKETDTVYEIG
                     TGKGHLTTKLAKISKQVTSIELDSHLFNLSSEKLKLNIRVTLIHQDILQFQFPNKQRYK
                     IVGNIPYHLSTQIIKKVVFESHASDIYLIVEEGFYKRTLDIHRTLGLLLHTQVSIQQLL
                     KLPAECFHPKPKVNSVLIKLTRHTTDVPDKYWKLYTYFVSKWVNREYRQLFTKNQFHQA
                     MKHAKVNNLSTVTYEQVLSIFNSYLLFNGRK"
     gene            complement(4167..4928)
                     /gene="erm"
                     /label=erm
     rep_origin      5356..5944
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    6130..6270
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(6372..6563)
                     /codon_start=1
                     /gene="rop"
                     /product="ROP protein"
                     /label=rop
                     /note="regulatory protein; involved in determination of 
                     plasmid copy number"
                     /protein_id="AAK27829.1"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     gene            complement(6372..6563)
                     /gene="rop"
                     /label=rop
     CDS             complement(7201..8391)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline-resistance protein"
                     /label=tet
                     /protein_id="AAK27830.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            complement(7201..8391)
                     /gene="tet"
                     /label=tet
     promoter        complement(8439..8467)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     gene            complement(8683..10032)
                     /gene="phoZ"
                     /allele="phoZ2"
                     /label=phoZ
                     /note="alkaline phosphatase; derived from Enterococcus 
                     faecalis; expression is dependent upon integration event to
                     create in-frame start codon"
     CDS             10619..11413
                     /codon_start=1
                     /gene="aphA3"
                     /product="aminoglycoside phosphotransferase A3"
                     /label=aphA3
                     /note="confers kanamycin-resistance"
                     /protein_id="AAK27831.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            10619..11413
                     /gene="aphA3"
                     /label=aphA3
     CDS             complement(11817..12515)
                     /codon_start=1
                     /gene="repAts"
                     /product="temperature-sensitive RepA"
                     /label=repAts
                     /protein_id="AAK27832.1"
                     /translation="MAIKNTKARNFGFLLYPDSIPNDWKEKLESLGVSMAVSPLHDMDE
                     KKDKDTWNNSNIIQNGKHYKKPHYHVIYIARNPVTIESVRNKIKRKLGNSSVAHVEILD
                     YIKGSYEYLTHESKDAIAKNKHIYDKKDILNINDFDIDRYITLDESQKRELKNLLLDIV
                     DDYNLVNTKDLMAFIRLRGAEFGILNTNDVKDIVSTNSSAFRLWFEGNYQCGYRASYAK
                     VLDAETGEIK"
     gene            complement(11817..12515)
                     /gene="repAts"
                     /label=repAts
     CDS             13127..13138
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"