LOCUS       Exported                4939 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle vector pBS423 DNA, complete sequence.
ACCESSION   AB687999
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4939)
  AUTHORS   Hirayama Y, Sakanaka M, Fukuma H, Murayama H, Kano Y, Fukiya S, 
            Yokota A.
  TITLE     Development of a Double-Crossover Markerless Gene Deletion System in
            Bifidobacterium longum: Functional Analysis of the 
            alpha-Galactosidase Gene for Raffinose Assimilation
  JOURNAL   Appl. Environ. Microbiol. 78 (14), 4984-4994 (2012)
  PUBMED    22582061
REFERENCE   2  (bases 1 to 4939)
  AUTHORS   Hirayama Y, Sakanaka M, Fukuma H, Murayama H, Kano Y, Fukiya S, 
            Yokota A.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-DEC-2011) Contact:Satoru Fukiya Hokkaido University, 
            Laboratory of Microbial Physiology, Research Faculty of Agriculture;
            Kita 9, Nishi 9, Kita-ku, Sapporo, Hokkaido 060-8589, Japan
REFERENCE   3  (bases 1 to 4939)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4939
                     /organism="Shuttle vector pBS423"
                     /mol_type="other DNA"
                     /db_xref="taxon:1128650"
     misc_feature    2..2959
                     /note="derived from Bifidobacterium longum BK25 plasmid 
                     pTB4"
     CDS             complement(297..857)
                     /codon_start=1
                     /gene="pTB4_p1"
                     /product="hypothetical protein"
                     /label=pTB4_p1
                     /protein_id="BAL42606.1"
                     /translation="MTNQLLTPAQCPAHRRRIRDAVPGRVQAGHRAQRLGAGTGVRAPL
                     PLLCGRALPAHGREHPRQRDDAAQGAQGEGRWKMTFQKKEEPLPVRGRRHHPERSSTLM
                     RFPMRSDGYIHGHPHGHPHGHLHGHLHGHLHGHLHGHLHGHPHGDVHQIEGKQARRFMY
                     MNRARSLPRPSAQEQQPECRRPE"
     gene            complement(297..857)
                     /gene="pTB4_p1"
                     /label=pTB4_p1
     CDS             1010..1933
                     /codon_start=1
                     /gene="repA"
                     /product="plasmid replication initiation protein RepA"
                     /label=repA
                     /protein_id="BAL42607.1"
                     /translation="MSNEYVQGTLELTRAFDGWWLPQRPLCCDDDYSQLVRRSRIDALR
                     CKHIEANPAAQVNMLVVDVDSSEGRLMSLWGHRDMPPNFIAENPANGHCHAGWVLTEPV
                     CRTDMARLKPLKLLHAVTEGLRRSVDGDEGYSGLLMKNPLSDAWDSDLCREDTYDLPDL
                     VAALEAHGDMPPKSWTRTKRAREVGVGRNCTLFDEARTLAYREVRRLPDRTPASSDLLR
                     EYVRRTCHEINASFPDPLPVREVNDTAKSIHKWITTRSRMWRDGAVANAATFVAIQSAR
                     GKKSGEARQNAFEEKFAQYAQEVLGQ"
     gene            1010..1933
                     /gene="repA"
                     /label=repA
     misc_feature    1196..1744
                     /gene="repA"
                     /note="deleted in the conditional replication vector 
                     pBS423-delta repA"
     CDS             1930..2220
                     /codon_start=1
                     /gene="repB"
                     /product="plasmid replication initiation protein RepB"
                     /label=repB
                     /protein_id="BAL42608.1"
                     /translation="MTIQTIRKKRPLPAKELAEAYGVSVRTIKYWNSQTREDWIDEQAT
                     LRESIRAYHDDDGHSWSQTAEHFNMTQGAVRQRAYRARKEREAEAKAARPE"
     gene            1930..2220
                     /gene="repB"
                     /label=repB
     CDS             complement(2421..2702)
                     /codon_start=1
                     /gene="pTB4_p2"
                     /product="hypothetical protein"
                     /label=pTB4_p2
                     /note="Similar to Blon_0217 (Ribbon-helix-helix protein, 
                     copG family) [Bifidobacterium longum subsp. infantis ATCC 
                     15697]"
                     /protein_id="BAL42609.1"
                     /translation="MSDFSERDRQLMALVGLTEEQVLEDERMAESETVPDDLTGRVYYG
                     LHLAEPDEQMVSVSVRMPKSTLDGLTAMARRYHISRSEYMRRKLAGAI"
     gene            complement(2421..2702)
                     /gene="pTB4_p2"
                     /label=pTB4_p2
     CDS             complement(2699..2941)
                     /codon_start=1
                     /gene="pTB4_p3"
                     /product="hypothetical protein"
                     /label=pTB4_p3
                     /protein_id="BAL42610.1"
                     /translation="MERHPELSEKDVVTAFRSVMVDAERESGAWMAIGLDGRGRNVEML
                     YRVVGDLVVIYHAFTPPTKKFRREIDRLRGDGRTL"
     gene            complement(2699..2941)
                     /gene="pTB4_p3"
                     /label=pTB4_p3
     misc_feature    2960..3788
                     /label=derived from Escherichia coli K12 plasmid pUC18
                     /note="derived from Escherichia coli K12 plasmid pUC18"
     rep_origin      complement(3042..3630)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    3789..1
                     /note="derived from Escherichia coli-Bifidobacterium 
                     shuttle vector pBRASTA101"
     CDS             complement(3852..4526)
                     /codon_start=1
                     /gene="aad(9)"
                     /product="spectinomycin adenyltransferase"
                     /function="Spectinomycin resistance"
                     /label=aad(9)
                     /protein_id="BAL42611.1"
                     /translation="MFGSGVESGLKPNSDLDFLVVVSEPLTDQSKEILIQKIRPISKKI
                     GDKSNLRYIELTIIIQQEMVPWNHPPKQEFIYGEWLQELYEQGYIPQKELNSDLTIMLY
                     QAKRKNKRIYGNYDLEELLPDIPFSDVRRAIMDSSEELIDNYQDDETNSILTLCRMILT
                     MDTGKIIPKDIAGNAVAESSPLEHRERILLAVRSYLGENIEWTNENVNLTINYLNNRLK
                     KL"
     gene            complement(3852..4526)
                     /gene="aad(9)"
                     /label=aad(9)