LOCUS       Exported                9376 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Plasmid vector pMD.SD.Gad, complete sequence.
ACCESSION   KJ870101
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9376)
  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 9376)
  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 9376)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9376
                     /organism="Plasmid vector pMD.SD.Gad"
                     /mol_type="other DNA"
                     /db_xref="taxon:1622226"
     CDS             411..1205
                     /codon_start=1
                     /product="Kan"
                     /label=Kan
                     /protein_id="AJS13704.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     protein_bind    complement(1396..1429)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             1500..1973
                     /codon_start=1
                     /product="tviD"
                     /label=tviD
                     /protein_id="AJS13705.1"
                     /translation="MSITLSFQNASMNKLFEKECRNVATRALKYVRQKKTEGRLDEALS
                     VLISLKRIEPDVSRLMREYKQIIRLFNESRKDGGSTITSYEHLDYAKKLLVFDSENAYA
                     LKYAALNAMHLRDYTQALQYWQRLEKVNGPTEPVTRQISTCITALQKNTSGKS"
     CDS             complement(2040..2918)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        2945..3229
                     /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             3257..4657
                     /codon_start=1
                     /product="gadA"
                     /label=gadA
                     /protein_id="AJS13706.1"
                     /translation="MDQKLLTDFRSELLDSRFGAKAISTIAESKRFPLHEMRDDVAFQI
                     INDELYLDGNARQNLATFCQTWDDENVHKLMDLSINKNWIDKEEYPQSAAIDLRCVNMV
                     ADLWHAPAPKNGQAVGTNTIGSSEACMLGGMAMKWRWRKRMEAAGKPTDKPNLVCGPVQ
                     ICWHKFARYWDVELREIPMRPGQLFMDPKRMIEACDENTIGVVPTFGVTYTGNYEFPQP
                     LHDALDKFQADTGIDIDMHIDAASGGFLAPFVAPDIVWDFRLPRVKSISASGHKFGLAP
                     LGCGWVIWRDEEALPQELVFNVDYLGGQIGTFAINFSRPAGQVIAQYYEFLRLGREGYT
                     KVQNASYQVAAYLADEIAKQGPYEFICTGRPDEGIPAVCFKLKDGEDPGYTLYDLSERL
                     RLRGWQVPAFTLGGEATDIVVMRIMCRRGFEMDFAELLLEDYKASLKYLSDHPKLQGIA
                     QQNSFKHT"
     CDS             4676..6106
                     /codon_start=1
                     /product="gadB"
                     /label=gadB
                     /protein_id="AJS13707.1"
                     /translation="MRSNHFKEFKMDKKQVTDLRSELLDSRFGAKSISTIAESKRFPLH
                     EMRDDVAFQIINDELYLDGNARQNLATFCQTWDDENVHKLMDLSINKNWIDKEEYPQSA
                     AIDLRCVNMVADLWHAPAPKNGQAVGTNTIGSSEACMLGGMAMKWRWRKRMEAAGKPTD
                     KPNLVCGPVQICWHKFARYWDVELREIPMRPGQLFMDPKRMIEACDENTIGVVPTFGVT
                     YTGNYEFPQPLHDALDKFQADTGIDIDMHIDAASGGFLAPFVAPDIVWDFRLPRVKSIS
                     ASGHKFGLAPLGCGWVIWRDEEALPQELVFNVDYLGGQIGTFAINFSRPAGQVIAQYYE
                     FLRLGREGYTKVQNASYQVAAYLADEIAKLGPYEFICTGRPDEGIPAVCFKLKDGEDPG
                     YTLYDLSERLRLRGWQVPAFTLGGEATDIVVMRIMCRRGFEMDFAELLLEDYKASLKYL
                     SDHPKLQGIAQQNSFKHT"
     CDS             6128..7663
                     /codon_start=1
                     /product="gadC"
                     /label=gadC
                     /protein_id="AJS13708.1"
                     /translation="MATSVQTGKAKQLTLLGFFAITASMVMAVYEYPTFATSGFSLVFF
                     LLLGGILWFIPVGLCAAEMATVDGWEEGGVFAWVSNTLGPRWGFAAISFGYLQIAIGFI
                     PMLYFVLGALSYILKWPALNEDPITKTIAALIILWALALTQFGGTKYTARIAKVGFFAG
                     ILLPAFILIALAAIYLHSGAPVAIEMDSKTFFPDFSKVGTLVVFVAFILSYMGVEASAT
                     HVNEMSNPGRDYPLAMLLLMVAAICLSSVGGLSIAMVIPGNEINLSAGVMQTFTVLMSH
                     VAPEIEWTVRVISALLLLGVLAEIASWIVGPSRGMYVTAQKNLLPAAFAKMNKNGVPVT
                     LVISQLVITSIALIILTNTGGGNNMSFLIALALTVVIYLCAYFMLFIGYIVLVLKHPDL
                     KRTFNIPGGKGVKLVVAIVGLLTSIMAFIVSFLPPDNIQGDSTDMYVELLVVSFLVVLA
                     LPFILYAVHDRKGKANTGVTLEPINSQNAPKGHFFLHPRARSPHYIVMNDKKH"
     protein_bind    complement(7709..7756)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(7947..8318)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            8351..8460
                     /note="incP origin of transfer"
     rep_origin      8620..9008
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     terminator      9144..9230
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      9322..9349
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"