LOCUS       Exported                3623 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Broad host range vector vector pCVD002, complete sequence.
ACCESSION   KM017937
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3623)
  AUTHORS   Taton A, Unglaub F, Wright NE, Zeng WY, Paz-Yepez J, Brahamsha B, 
            Palenik B, Peterson TC, Haerizadeh F, Golden SS, Golden JW.
  TITLE     Broad-host-range vector system for synthetic biology and 
            biotechnology in cyanobacteria
  JOURNAL   Nucleic Acids Res. (2014) In press
  PUBMED    25074377
REFERENCE   2  (bases 1 to 3623)
  AUTHORS   Taton A, Unglaub F, Wright N, Zeng WY, Paz Yepes J, Brahamsha B, 
            Palenik B, Peterson T, Haerizadeh F, Golden SS, Golden JW.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-JUN-2014) Division of Biological Sciences, University 
            of California, San Diego, 9500 Gilman Dr., La Jolla, CA 92093, USA
REFERENCE   3  (bases 1 to 3623)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3623
                     /organism="Broad host range vector vector pCVD002"
                     /mol_type="other DNA"
                     /note="aadA is a device that confers spectinomycin and 
                     streptomycin (SpSm) resistance The stop codon was erased by
                     mistake, another stop codon is however located 15 nt 
                     downstream in the transcriptional terminator This device 
                     works perfectly well"
                     /db_xref="taxon:1531860"
     primer_bind     293..309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        320..338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    370..375
                     /label=EcoRV
                     /note="EcoRV"
     misc_feature    complement(373..1648)
                     /note="CVD-ANTR; device for the GC-adaptor assembly: 
                     antibiotic resistance markers"
     misc_feature    complement(373..393)
                     /note="C2G; GC-adaptor for seamless assembly, flanking 
                     antibiotic resistance markers, functional devices or custom
                     sequences; synthetic"
     misc_feature    394..399
                     /label=AgeI
                     /note="AgeI"
     regulatory      406..789
                     /regulatory_class="promoter"
                     /note="PaadA; upstream region of aadA ORF including 
                     promoter and RBS; derived from pRL1383a in INSD accession 
                     AF403426, originally derived from the omega interposon INSD
                     accession M60473"
     misc_feature    790..1580
                     /note="similar to aadA; confers spectinomycin and/or 
                     streptomycin resistance; aminoglycoside-3:adenyltransferase
                     fusion; derived from pRL1383a in INSD accession AF403426, 
                     originally derived from the omega interposon INSD accession
                     M60473; stop codon was erased by mistake however as part of
                     the assembly device CVD002/CVD-aadA, a stop codon is 
                     located 15 nucleotides downstream"
     CDS             790..1575
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVG"
     regulatory      1586..1612
                     /regulatory_class="terminator"
                     /note="trpA T; trpA terminator from Lorist6 INSD accession 
                     X98450, one nt shorter than reference; Lorist6"
     misc_feature    1622..1627
                     /label=NheI
                     /note="NheI"
     misc_feature    complement(1628..1648)
                     /note="GC; GC-adaptor for seamless assembly, flanking 
                     antibiotic resistance markers or Escherichia coli origins 
                     to be assembled with a cyanobacterial replicon; synthetic"
     misc_feature    1646..1651
                     /label=EcoRV
                     /note="EcoRV"
     rep_origin      complement(1817..2490)
                     /direction=LEFT
                     /note="colE1_pMA-RQ; origin of replication of colE1 found 
                     inderived from pMA-RQ GeneART vector"
     rep_origin      complement(1876..2464)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2635..3495)
                     /codon_start=1
                     /product="bla_pMA-RQ"
                     /label=bla_pMA-RQ
                     /note="confers ampicillin resistance; beta-lactamase; 
                     derived from pMA-RQ GeneART vector, has one point mutation 
                     compared to pUC19"
                     /protein_id="AII99781.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3496..3600)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      complement(3496..3565)
                     /regulatory_class="promoter"
                     /note="Pbla; upstream region of the bla ORF including 
                     promoter and RBS; derived from pUC19"