LOCUS       Exported                4658 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBAMD1-2, complete sequence.
ACCESSION   KM403113
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4658)
  AUTHORS   Martinez-Garcia E, Aparicio T, de Lorenzo V, Nikel PI.
  TITLE     New transposon tools tailored for metabolic engineering of 
            gram-negative microbial cell factories
  JOURNAL   Front Bioeng Biotechnol 2, 46 (2014)
  PUBMED    25389526
REFERENCE   2  (bases 1 to 4658)
  AUTHORS   Martinez-Garcia E, Aparicio T, de Lorenzo V, Nikel PI.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-AUG-2014) Systems Biology Program, Centro Nacional de 
            Biotecnologia (CNB-CSIC), C/Darwin 3, Madrid, Madrid 28049, Spain
REFERENCE   3  (bases 1 to 4658)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4658
                     /organism="Cloning vector pBAMD1-2"
                     /mol_type="other DNA"
                     /db_xref="taxon:1572690"
     misc_feature    10..28
                     /label=mosaic element I (ME-I)
                     /note="mosaic element I (ME-I)"
     misc_feature    62..118
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     regulatory      127..156
                     /regulatory_class="terminator"
                     /note="terminator T500"
     CDS             266..1081
                     /codon_start=1
                     /gene="aphA"
                     /product="AphA"
                     /label=aphA
                     /note="aminoglycoside 3-phosphotransferase; kanamycin 
                     resistance"
                     /protein_id="AIX94012.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            266..1081
                     /gene="aphA"
                     /label=aphA
     regulatory      1092..1163
                     /regulatory_class="terminator"
                     /note="terminator of gene 32 (T32)"
     misc_feature    1174..1192
                     /label=mosaic element O (ME-O)
                     /note="mosaic element O (ME-O)"
     regulatory      1199..1301
                     /regulatory_class="terminator"
                     /note="terminator T0"
     terminator      1203..1297
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1402..2832
                     /codon_start=1
                     /gene="tnpA"
                     /product="TnpA"
                     /label=tnpA
                     /note="Tn5 transposase"
                     /protein_id="AIX94013.1"
                     /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
                     ITISSEGSKAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
                     HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
                     KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
                     YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
                     VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
                     QRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTP
                     DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
                     KLDGFLAAKDLMAQGIKI"
     gene            1402..2832
                     /gene="tnpA"
                     /label=tnpA
     CDS             1402..2829
                     /codon_start=1
                     /product="transposase from the bacterial Tn5 transposon 
                     (Reznikoff, 1993)"
                     /label=Tn5 transposase
                     /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
                     ITISSEGSKAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
                     HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
                     KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
                     YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
                     VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
                     QRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTP
                     DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
                     KLDGFLAAKDLMAQGIKI"
     promoter        2886..2977
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2978..3838
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AIX94014.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            2978..3838
                     /gene="bla"
                     /label=bla
     terminator      3842..3881
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     misc_feature    complement(3894..4139)
                     /label=oriT
                     /note="oriT"
     oriT            4029..4137
                     /note="incP origin of transfer"
     misc_feature    complement(4148..4539)
                     /label=oriR6K
                     /note="oriR6K"
     rep_origin      4148..4536
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     regulatory      complement(4554..4658)
                     /regulatory_class="terminator"
                     /note="terminator T1"
     terminator      4567..4653
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"