LOCUS       Exported                8365 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pTBR101-CM DNA, complete sequence.
ACCESSION   AB688000
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8365)
  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 8365)
  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 8365)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8365
                     /organism="Shuttle vector pTBR101-CM"
                     /mol_type="other DNA"
                     /db_xref="taxon:1128651"
     misc_feature    7..1044
                     /label=derived from Staphylococcus aureus plasmid pC194
                     /note="derived from Staphylococcus aureus plasmid pC194"
     CDS             complement(106..756)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /function="chloramphenicol resistance"
                     /label=cat
                     /protein_id="BAL42612.1"
                     /translation="MNFNKIDLDNWKRKEIFNHYLNQQTTFSITTEIDISVLYRNIKQE
                     GYKFYPAFIFLVTRVINSNTAFRTGYNSDGELGYWDKLEPLYTIFDGVSKTFSGIWTPV
                     KNDFKEFYDLYLSDVEKYNGSGKLFPKTPIPENAFSLSIIPWTSFTGFNLNINNNSNYL
                     LPIITAGKFINKGNSIYLPLSLQVHHSVCDGYHAGLFMNSIQELSDRPNDWLL"
     gene            complement(106..756)
                     /gene="cat"
                     /label=cat
     misc_feature    1046..1393
                     /label=derived from Escherichia coli plasmid pBR322
                     /note="derived from Escherichia coli plasmid pBR322"
     misc_feature    1104..1393
                     /gene="tetA"
                     /note="tetA was disrupted by the insertion of 
                     Bifidobacterium longum BK25 plasmid pTB4"
     gene            1104..1393
                     /gene="tetA"
                     /label=tetA
     misc_feature    1394..4351
                     /note="derived from Bifidobacterium longum BK25 plasmid 
                     pTB4"
     CDS             complement(1689..2249)
                     /codon_start=1
                     /gene="pTB4_p1"
                     /product="hypothetical protein"
                     /label=pTB4_p1
                     /protein_id="BAL42613.1"
                     /translation="MTNQLLTPAQCPAHRRRIRDAVPGRVQAGHRAQRLGAGTGVRAPL
                     PLLCGRALPAHGREHPRQRDDAAQGAQGEGRWKMTFQKKEEPLPVRGRRHHPERSSTLM
                     RFPMRSDGYIHGHPHGHPHGHLHGHLHGHLHGHLHGHLHGHPHGDVHQIEGKQARRFMY
                     MNRARSLPRPSAQEQQPECRRPE"
     gene            complement(1689..2249)
                     /gene="pTB4_p1"
                     /label=pTB4_p1
     CDS             2402..3325
                     /codon_start=1
                     /gene="repA"
                     /product="plasmid replication initiation protein RepA"
                     /label=repA
                     /protein_id="BAL42614.1"
                     /translation="MSNEYVQGTLELTRAFDGWWLPQRPLCCDDDYSQLVRRSRIDALR
                     CKHIEANPAAQVNMLVVDVDSSEGRLMSLWGHRDMPPNFIAENPANGHCHAGWVLTEPV
                     CRTDMARLKPLKLLHAVTEGLRRSVDGDEGYSGLLMKNPLSDAWDSDLCREDTYDLPDL
                     VAALEAHGDMPPKSWTRTKRAREVGVGRNCTLFDEARTLAYREVRRLPDRTPASSDLLR
                     EYVRRTCHEINASFPDPLPVREVNDTAKSIHKWITTRSRMWRDGAVANAATFVAIQSAR
                     GKKSGEARQNAFEEKFAQYAQEVLGQ"
     gene            2402..3325
                     /gene="repA"
                     /label=repA
     CDS             3322..3612
                     /codon_start=1
                     /gene="repB"
                     /product="plasmid replication initiation protein RepB"
                     /label=repB
                     /protein_id="BAL42615.1"
                     /translation="MTIQTIRKKRPLPAKELAEAYGVSVRTIKYWNSQTREDWIDEQAT
                     LRESIRAYHDDDGHSWSQTAEHFNMTQGAVRQRAYRARKEREAEAKAARPE"
     gene            3322..3612
                     /gene="repB"
                     /label=repB
     CDS             complement(3813..4094)
                     /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="BAL42616.1"
                     /translation="MSDFSERDRQLMALVGLTEEQVLEDERMAESETVPDDLTGRVYYG
                     LHLAEPDEQMVSVSVRMPKSTLDGLTAMARRYHISRSEYMRRKLAGAI"
     gene            complement(3813..4094)
                     /gene="pTB4_p2"
                     /label=pTB4_p2
     CDS             complement(4091..4333)
                     /codon_start=1
                     /gene="pTB4_p3"
                     /product="hypothetical protein"
                     /label=pTB4_p3
                     /protein_id="BAL42617.1"
                     /translation="MERHPELSEKDVVTAFRSVMVDAERESGAWMAIGLDGRGRNVEML
                     YRVVGDLVVIYHAFTPPTKKFRREIDRLRGDGRTL"
     gene            complement(4091..4333)
                     /gene="pTB4_p3"
                     /label=pTB4_p3
     misc_feature    4352..1
                     /label=derived from Escherichia coli plasmid pBR322
                     /note="derived from Escherichia coli plasmid pBR322"
     misc_feature    4352..5252
                     /gene="tetA"
                     /note="tetA was disrupted by the insertion of 
                     Bifidobacterium longum BK25 plasmid pTB4"
     gene            4352..5252
                     /gene="tetA"
                     /label=tetA
     CDS             5891..6082
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    6184..6324
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(6510..7098)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7269..8129)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="Ampicillin resistance"
                     /protein_id="BAL42618.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(7269..8129)
                     /gene="bla"
                     /label=bla
     promoter        complement(8130..8234)
                     /gene="bla"
                     /label=AmpR promoter