LOCUS       Exported                5023 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBAM1-GFP, complete sequence.
ACCESSION   HQ908072
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5023)
  AUTHORS   Martinez-Garcia E, Calles B, Arevalo-Rodriguez M, de Lorenzo V.
  TITLE     pBAM1: an all-synthetic genetic tool for analysis and construction 
            of complex bacterial phenotypes
  JOURNAL   BMC Microbiol. 11 (1), 38 (2011)
  PUBMED    21342504
REFERENCE   2  (bases 1 to 5023)
  AUTHORS   Martinez-Garcia E, Calles B, Arevalo-Rodriguez M, de Lorenzo V.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-JAN-2011) Systems Biology Program, Centro Nacional de 
            Biotecnologia, C/ Darwin 3, Madrid, Madrid 28049, Spain
REFERENCE   3  (bases 1 to 5023)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5023
                     /organism="Cloning vector pBAM1-GFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:990067"
     rep_origin      9..400
                     /label=R6KoriV
                     /note="R6KoriV"
     rep_origin      12..400
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     misc_feature    419..662
                     /note="oriT; origin of transference"
     oriT            419..527
                     /note="incP origin of transfer"
     terminator      674..713
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(717..1577)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="ADY68347.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(717..1577)
                     /gene="bla"
                     /label=bla
     promoter        complement(1578..1669)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(1723..3153)
                     /codon_start=1
                     /gene="tnpA"
                     /product="Tn5 hyperactive transposase"
                     /label=tnpA
                     /note="contains substitutions E54K and L372P"
                     /protein_id="ADY68346.1"
                     /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
                     ITISSEGSKAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
                     HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
                     KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
                     YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
                     VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
                     QRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTP
                     DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
                     KLDGFLAAKDLMAQGIKI"
     gene            complement(1723..3153)
                     /gene="tnpA"
                     /label=tnpA
     CDS             complement(1726..3153)
                     /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"
     misc_feature    3247..3265
                     /note="ME-I; mosaic element"
     CDS             3274..3984
                     /codon_start=1
                     /gene="gfp"
                     /product="GFP"
                     /label=gfp
                     /note="lacks start and stop codons"
                     /protein_id="ADY68349.1"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     gene            3274..3984
                     /gene="gfp"
                     /label=gfp
     misc_difference 3945
                     /gene="gfp"
                     /replace="a"
     gene            4102..4920
                     /gene="neo"
                     /label=neo
     CDS             4102..4917
                     /codon_start=1
                     /gene="neo"
                     /product="kanamycin resistance protein"
                     /label=neo
                     /note="originates from transposon Tn903"
                     /protein_id="ADY68348.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     misc_feature    5002..5020
                     /note="ME-O; mosaic element"