LOCUS       Exported                8082 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Promoter screenings vector pMM223, complete sequence.
ACCESSION   AF076212
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8082)
  AUTHORS   Vriesema AJ, Brinkman R, Kok J, Dankert J, Zaat SA.
  TITLE     Broad-host-range shuttle vectors for screening of regulated promoter
            activity in viridans group streptococci: isolation of a pH-regulated
            promoter
  JOURNAL   Appl. Environ. Microbiol. 66 (2), 535-542 (2000)
  PUBMED    10653715
REFERENCE   2  (bases 1 to 8082)
  AUTHORS   Vriesema AJM., Brinkman R, Kok J, Dankert J, Zaat SAJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-JUL-1998) Medical Microbiology, Academic Medical 
            Center, University of Amsterdam, Meibergdreef 15, Amsterdam 1105 AZ,
            The Netherlands
REFERENCE   3  (bases 1 to 8082)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8082
                     /organism="Promoter screenings vector pMM223"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:79019"
     regulatory      26..68
                     /regulatory_class="terminator"
     misc_feature    100..168
                     /note="multiple cloning site; MCS"
     regulatory      184..189
                     /regulatory_class="ribosome_binding_site"
     CDS             196..948
                     /codon_start=1
                     /gene="aad(9)"
                     /product="spectinomycin adenyltransferase AAD(9)"
                     /label=aad(9)
                     /protein_id="AAD46051.1"
                     /translation="MNTYEQINKVKKILRKHLKNNLIGTYMFGSGVESGLKPNSDLDFL
                     VVVSEPLTDQSKEILIQKIRPISKKIGDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYG
                     EWLQELYEQGYIPQKELNSDLTIMLYQAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMD
                     SSEELIDNYQDDETNSILTLCRMILTMDTGKIIPKDIAGNAVAESSPLEHRERILLAVR
                     SYLGENIEWTNENVNLTINYLNNRLKKL"
     gene            196..948
                     /gene="aad(9)"
                     /label=aad(9)
                     /note="promoterless"
     misc_feature    959..4460
                     /label=trp-lacZ fusion
                     /note="trp-lacZ fusion"
     primer_bind     1193..1209
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1194..1197
                     /regulatory_class="ribosome_binding_site"
     CDS             1205..4279
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-d-galactosidase"
                     /label=lacZ
                     /protein_id="AAD46052.1"
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYG
                     QDSRLPSEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQ
                     ISDFHVATRFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIID
                     ERGGYADRVTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIEN
                     GLLLLNGKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLW
                     YTLCDRYGLYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWS
                     LGNESGHGANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVP
                     KWSIKKWLSLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSL
                     IKYDENGNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQ
                     TIEVTSEYLFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQ
                     LWLTVRVVQPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELG
                     NKRWQFNRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWK
                     AAGHYQAEAALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDV
                     EVASDTPHPARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPY
                     VFPSENGLRCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDG
                     FHMGIGGDDSWSPSVSAELQLSAGRYHYQLVWCQK"
     gene            1205..4279
                     /gene="lacZ"
                     /label=lacZ
                     /note="promoterless"
     primer_bind     complement(1225..1241)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(5365..6063)
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein RepA"
                     /label=repA
                     /protein_id="AAD46053.1"
                     /translation="MAIKNTKARNFGFLLYPDSIPNDWKEKLESLGVSMAVSPLHDMDE
                     KKDKDTWNSSDVIRNGKHYKKPHYHVIYIARNPVTIESVRNKIKRKLGNSSVAHVEILD
                     YIKGSYEYLTHESKDAIAKNKHIYDKKDILNINDFDIDRYITLDESQKRELKNLLLDIV
                     DDYNLVNTKDLMAFIRLRGAEFGILNTNDVKDIVSTNSSAFRLWFEGNYQCGYRASYAK
                     VLDAETGEIK"
     gene            complement(5365..6063)
                     /gene="repA"
                     /label=repA
     CDS             6675..6686
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             7156..7215
                     /codon_start=1
                     /product="emrC leader peptide"
                     /label=emrC leader peptide
                     /protein_id="AAD46054.1"
                     /translation="MGIFSIFVISTVHYQPNKK"
     CDS             7275..8009
                     /codon_start=1
                     /gene="emrC"
                     /product="adenine methylase"
                     /label=emrC
                     /protein_id="AAD46055.1"
                     /translation="MNEKNIKHSQNFITSKHNIDKIMTNIRLNEHDNIFEIGSGKGHFT
                     LELVKRCNFVTAIEIDHKLCKTTENKLVDHDNFQVLNKDIMQFKFPKNQSYKIYGNIPY
                     NISTDIIRKIVFDSIANEIYLIVEYGFAKRLLNTKRSLALLLMAEVDISILSMVPREYF
                     HPKPKVNSSLIRLSRKKSRISHKDKQKYNYFVMKWVNKEYKKIFTKNQFNNSLKHAGID
                     DLNNISFEQFLSLFNSYKLFNK"
     gene            7275..8009
                     /gene="emrC"
                     /label=emrC