LOCUS       Exported                9278 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pG106, complete sequence.
ACCESSION   MF480755
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9278)
  AUTHORS   Jones KR, Macrina FL, Lewis JP.
  TITLE     Porphyromonas gingivalis shuttle vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9278)
  AUTHORS   Jones KR, Macrina FL, Lewis JP.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-JUL-2017) OCMB, Virginia Commonwealth University, 
            Philips Institute, 521 North 11th Street, Richmond, VA 23298, USA
REFERENCE   3  (bases 1 to 9278)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9278
                     /organism="Shuttle vector pG106"
                     /mol_type="other DNA"
                     /label=Porphyromonas gingivalis vector
                     /note="Porphyromonas gingivalis vector"
                     /db_xref="taxon:2045299"
     RBS             316..324
                     /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             767..2359
                     /codon_start=1
                     /gene="repB"
                     /product="RepB"
                     /label=repB
                     /note="putative protein required for plasmid replication in
                     Porphyromonas gingivalis"
                     /protein_id="AVN57617.1"
                     /translation="MRTFEYRSLKPFEFFQIIFGSCDHNDPQCPIYPFEKELCQEYIDW
                     RAQQAGRKSKSKAGKKAIYNPAQPFRYNALVQAVYSVEPDAGRANDNGRRSKTWVVVGD
                     ATSEDDIKVCGWDSLEELMGRYKFVVTTPATYVGRRRFKSNARYLYALAFDLDEVGVPE
                     MSEVIHQQTIGMTPQANIIVNSGDGIHLYYILAKPMPLYPKVYETLTNIKKALTRLIWN
                     KASSRTGEDNQVQIQPIIQPFRVPGSRTKHGDIVTAWHNADAPLHTIEELNRFASKPTL
                     KYLNSGVSQEQAQAFGQRGILPRIWLTKARAKELYPDWYQKRIVEGAPRKTWHVHRGLY
                     EWWLHMLWTNEKVAVGHRYHCIMFLAVYAKKCNVPFDILKRDALELVPRMEALTGNTGR
                     HFSVQDALDALKAYKESSETYPRQLIEDRSGLRIEPNKRNGRPQDLHLKLARGSRDILQ
                     EAKGRKDWREGNGRPKGSIVLAEDSPQYAKVQEWRDRNPNSNNKSRCARETGLSRPTRP
                     QVVAKPLGNALRT"
     gene            767..2359
                     /gene="repB"
                     /label=repB
     CDS             complement(2518..3495)
                     /codon_start=1
                     /gene="mob"
                     /product="Mob"
                     /label=mob
                     /note="putative protein required for plasmid stability in 
                     Porphyromonas gingivalis"
                     /protein_id="AVN57618.1"
                     /translation="MSKTSIHIEPCKIGSSEQHNQRLKHLDYVRPDLSPQNESWVGVAD
                     LPAHLEQLRILVKEKTGRAMQQKATPIREGVIVINQGTSINQLRGLATAIELRWGIKTL
                     QIYTHKDEGHIDSDDTWKPNLHAHMVLDWVNHDTGKSIKLSKQDMAEMQTMVADCLQMV
                     RGESSDVKHLGAIQYKNQAEELRLQTLKEQTMKEEQARELALAEAKRMEQVASEQIIAE
                     NKKLGGLWMDWQKSFEASEKRRESITARKIEEDKLWEELGLKGAFKSFWEECKDWFRGF
                     GDGLKDLALLIWASPRGVMDKDVRCTADRKRGKLLLKWAYTRSR"
     gene            complement(2518..3495)
                     /gene="mob"
                     /label=mob
     CDS             complement(4510..5043)
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="orfE; hypothetical protein"
                     /protein_id="AVN57619.1"
                     /translation="MLVRGAEPMEKRQQRGLFTVPGLLLAFCSHGSSIADGLVAHPSEG
                     FYIIYTTSSEEFLTLSEEDYSLFASFANLVVREDSNSNGLRSAKPSGDGWKFGGKGKGK
                     LDAIHIGYELSKILGENQDFEIYVEYNGDGSYTVWYREVQKSSKGSNPVKTFNVLIRAN
                     EREGHTVSLFQPYA"
     rep_origin      complement(5013..5601)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5939..6676)
                     /codon_start=1
                     /gene="ermAM"
                     /product="ErmAM"
                     /label=ermAM
                     /note="erythromycin resistance"
                     /protein_id="AVN57620.1"
                     /translation="MNKNIKYSQNFLTSEKVLNQIIKQLNLKETDTVYEIGTGKGHLTT
                     KLAKISKQVTSIELDSHLFNLSSEKLKLNTRVTLIHQDILQFQFPNKQRYKIVGSIPYH
                     LSTQIIKKVVFESHASDIYLIVEEGFYKRTLDIHRTLGLLLHTQVSIQQLLKLPAECFH
                     PKPKVNSVLIKLTRHTTDVPDKYWKLYTYFVSKWVNREYRQLFTKNQFHQAMKHAKVNN
                     LSTVTYEQVLSIFNSYLLFNGRK"
     gene            complement(5939..6676)
                     /gene="ermAM"
                     /label=ermAM
     CDS             complement(6912..7712)
                     /codon_start=1
                     /gene="ermF"
                     /product="ErmF"
                     /label=ermF
                     /note="erythromycin resistance in Bacteroidetes"
                     /protein_id="AVN57621.1"
                     /translation="MTKKKLPVRFTGQHFTIDKVLIKDAIRQANISNQDTVLDIGAGKG
                     FLTVHLLKIANNVVAIENDTALVEHLRKLFSDARNVQVVGCDFRNFAVPKFPFKVVSNI
                     PYGITSDIFKILMFESLGNFLGGSIVLQLEPTQKLFSRKLYNPYTVFYHTFFDLKLVYE
                     VGPESFLPPPTVKSALLNIKRKHLFFDFKFKAKYLAFISCLLEKPDLSVKTALKSIFRK
                     SQVRSISEKFGLNLNAQIVCLSPSQWLNCFLEMLEVVPEKFHPS"
     gene            complement(6912..7712)
                     /gene="ermF"
                     /label=ermF
     oriT            8269..8378
                     /note="incP origin of transfer"
     CDS             complement(8620..8937)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRI?MVHSQYNL
                     L"
     CDS             8654..9010
                     /codon_start=1
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="orfG; codes for LacOPZ"
                     /protein_id="AVN57622.1"
                     /translation="MRCEIPHRCVRRKYRIRRHSPFRLRNCWEGRSVRASSLLRQLAKG
                     GCAARRLSWVTPGFSQSRRCKTTASEFELGTRGSSRVDLQACKLGVIMVIAVSCVKLLS
                     AHNSTQHTSRKHKV"
     primer_bind     8847..8863
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    8864..8920
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(8933..8949)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8957..8973
                     /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(8981..9011)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9026..9047
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."