LOCUS       Exported                9175 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Plasmid vector pGad, complete sequence.
ACCESSION   KJ870100
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9175)
  AUTHORS   Dharmasena MN, Stark CEC., Stibitz S, Kopecko DJ.
  TITLE     Acid-resistant, attenuated microbial vector for improved oral 
            delivery of multiple targeted antigens
  JOURNAL   Patent: US WO/2015/021147-B 12-FEB-2015; FDA-Center for Biologics 
            Evaluation and Research; NIH campus Bldg. 29, 8800 Rockville Pike; 
            Laboratory of Enteric and Sexually Transmitted Diseases; Bethesda, 
            MD; USA;
REFERENCE   2  (bases 1 to 9175)
  AUTHORS   Dharmasena MN, Stark CEC., Stibitz S, Kopecko DJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-MAY-2014) Laboratory of Enteric and Sexually 
            Transmitted Diseases, FDA-Center for Biologics Evaluation and 
            Research, NIH Campus Bldg. 29, 8800 Rockville Pike, Bethesda, MD 
            20892, USA
REFERENCE   3  (bases 1 to 9175)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9175
                     /organism="Plasmid vector pGad"
                     /mol_type="other DNA"
                     /db_xref="taxon:1622224"
     rep_origin      480..1025
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     CDS             1620..2435
                     /codon_start=1
                     /gene="aph(3)-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3050..3928)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3955..4239
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     CDS             4267..5667
                     /codon_start=1
                     /product="gadA"
                     /label=gadA
                     /protein_id="AJS13701.1"
                     /translation="MDQKLLTDFRSELLDSRFGAKAISTIAESKRFPLHEMRDDVAFQI
                     INDELYLDGNARQNLATFCQTWDDENVHKLMDLSINKNWIDKEEYPQSAAIDLRCVNMV
                     ADLWHAPAPKNGQAVGTNTIGSSEACMLGGMAMKWRWRKRMEAAGKPTDKPNLVCGPVQ
                     ICWHKFARYWDVELREIPMRPGQLFMDPKRMIEACDENTIGVVPTFGVTYTGNYEFPQP
                     LHDALDKFQADTGIDIDMHIDAASGGFLAPFVAPDIVWDFRLPRVKSISASGHKFGLAP
                     LGCGWVIWRDEEALPQELVFNVDYLGGQIGTFAINFSRPAGQVIAQYYEFLRLGREGYT
                     KVQNASYQVAAYLADEIAKQGPYEFICTGRPDEGIPAVCFKLKDGEDPGYTLYDLSERL
                     RLRGWQVPAFTLGGEATDIVVMRIMCRRGFEMDFAELLLEDYKASLKYLSDHPKLQGIA
                     QQNSFKHT"
     CDS             5686..7116
                     /codon_start=1
                     /product="gadB"
                     /label=gadB
                     /protein_id="AJS13702.1"
                     /translation="MRSNHFKEFKMDKKQVTDLRSELLDSRFGAKSISTIAESKRFPLH
                     EMRDDVAFQIINDELYLDGNARQNLATFCQTWDDENVHKLMDLSINKNWIDKEEYPQSA
                     AIDLRCVNMVADLWHAPAPKNGQAVGTNTIGSSEACMLGGMAMKWRWRKRMEAAGKPTD
                     KPNLVCGPVQICWHKFARYWDVELREIPMRPGQLFMDPKRMIEACDENTIGVVPTFGVT
                     YTGNYEFPQPLHDALDKFQADTGIDIDMHIDAASGGFLAPFVAPDIVWDFRLPRVKSIS
                     ASGHKFGLAPLGCGWVIWRDEEALPQELVFNVDYLGGQIGTFAINFSRPAGQVIAQYYE
                     FLRLGREGYTKVQNASYQVAAYLADEIAKLGPYEFICTGRPDEGIPAVCFKLKDGEDPG
                     YTLYDLSERLRLRGWQVPAFTLGGEATDIVVMRIMCRRGFEMDFAELLLEDYKASLKYL
                     SDHPKLQGIAQQNSFKHT"
     CDS             7138..8673
                     /codon_start=1
                     /product="gadC"
                     /label=gadC
                     /protein_id="AJS13703.1"
                     /translation="MATSVQTGKAKQLTLLGFFAITASMVMAVYEYPTFATSGFSLVFF
                     LLLGGILWFIPVGLCAAEMATVDGWEEGGVFAWVSNTLGPRWGFAAISFGYLQIAIGFI
                     PMLYFVLGALSYILKWPALNEDPITKTIAALIILWALALTQFGGTKYTARIAKVGFFAG
                     ILLPAFILIALAAIYLHSGAPVAIEMDSKTFFPDFSKVGTLVVFVAFILSYMGVEASAT
                     HVNEMSNPGRDYPLAMLLLMVAAICLSSVGGLSIAMVIPGNEINLSAGVMQTFTVLMSH
                     VAPEIEWTVRVISALLLLGVLAEIASWIVGPSRGMYVTAQKNLLPAAFAKMNKNGVPVT
                     LVISQLVITSIALIILTNTGGGNNMSFLIALALTVVIYLCAYFMLFIGYIVLVLKHPDL
                     KRTFNIPGGKGVKLVVAIVGLLTSIMAFIVSFLPPDNIQGDSTDMYVELLVVSFLVVLA
                     LPFILYAVHDRKGKANTGVTLEPINSQNAPKGHFFLHPRARSPHYIVMNDKKH"
     terminator      8906..8992
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      9084..9111
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"